title: Development of an assay to assess genotoxicity by particulate matter extract.
abstract: The current study describes a method for assessing the oxidative potential of common environmental stressors (ambient air particulate matter), using a plasmid relaxation assay where the extract caused single-strand breaks, easily visualised through electrophoresis. This assay utilises a miniscule amount (11 µg) of particulate matter (PM) extract compared to other, cell‑based methods (~3,000 µg). The negative impact of air pollution on human health has been extensively recognised. Among the air pollutants, PM plays an eminent role, as reflected in the broad scientific interest. PM toxicity highly depends on its composition (metals and organic compounds), which in turn has been linked to multiple health effects (such as cardiorespiratory diseases and cancer) through multiple toxicity mechanisms; the induction of oxidative stress is considered a major mechanism among these. In this study, the PM levels, oxidative potential, cytotoxicity and genotoxicity of PM in the region of Larissa, Greece were examined using the plasmid relaxation assay. Finally, coffee extracts from different varieties, derived from both green and roasted seeds, were examined for their ability to inhibit PM-induced DNA damage. These extracts also exerted an inhibitory effect on xanthine oxidase and catalase, but had no effect against superoxide dismutase. Overall, this study highlights the importance of assays for assessing the oxidative potential of widespread environmental stressors (PM), as well as the antioxidant capacity of beverages and food items, with the highlight being the development of a plasmid relaxation assay to assess the genotoxicity caused by PM using only a miniscule amount.
authors.affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, Larissa 41221, Greece.
authors.forename: Alexandros
authors.initials: A
authors.lastname: Priftis
authors.affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, Larissa 41221, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Papikinos
authors.affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, Larissa 41221, Greece.
authors.forename: Marina
authors.initials: M
authors.lastname: Koukoulanaki
authors.affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, Larissa 41221, Greece.
authors.forename: Efthalia
authors.initials: E
authors.lastname: Kerasioti
authors.affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, Larissa 41221, Greece.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Stagos
authors.affiliation: Coffee Island S.A., Patras 26334, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Konstantinopoulos
authors.affiliation: Laboratory of Clinical Virology, University of Crete, Medical School, Heraklion 71409, Greece.
authors.forename: Demetrios A
authors.initials: DA
authors.lastname: Spandidos
authors.affiliation: Aristotle University of Thessaloniki, Department of Chemical Engineering, Environmental Engineering Laboratory, Thessaloniki 54124, Greece.
authors.forename: Marianthi
authors.initials: M
authors.lastname: Kermenidou
authors.affiliation: Aristotle University of Thessaloniki, Department of Chemical Engineering, Environmental Engineering Laboratory, Thessaloniki 54124, Greece.
authors.forename: Spyros
authors.initials: S
authors.lastname: Karakitsios
authors.affiliation: Aristotle University of Thessaloniki, Department of Chemical Engineering, Environmental Engineering Laboratory, Thessaloniki 54124, Greece.
authors.forename: Dimosthenis
authors.initials: D
authors.lastname: Sarigiannis
authors.affiliation: Department of Forensic Sciences and Toxicology, Medical School, University of Crete, Heraklion 71003, Greece.
authors.forename: Aristides M
authors.initials: AM
authors.lastname: Tsatsakis
authors.affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, Larissa 41221, Greece.
authors.forename: Demetrios
authors.initials: D
authors.lastname: Kouretas
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D040503:Coffea / Q000737:chemistry; D053837:DNA Cleavage / Q000187:drug effects; D004249:DNA Damage; D006801:Humans; D009152:Mutagenicity Tests / Q000379:methods*; D052638:Particulate Matter / Q000633:toxicity*; D010936:Plant Extracts / Q000494:pharmacology; D059808:Polyphenols / Q000032:analysis
chemical_list: D000975:Antioxidants; D052638:Particulate Matter; D010936:Plant Extracts; D059808:Polyphenols