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Cytogenetic status of human lymphocytes after exposure to low concentrations of p,p'-DDT, and its metabolites (p,p'-DDE, and p,p'-DDD) in vitro
Authors:Geri? Marko  Ceraj-Ceri? Nikolina  Gajski Goran  Vasili? ?elimira  Capuder ?eljka  Garaj-Vrhovac Vera
Institution:a Mutagenesis Unit, Institute for Medical Research and Occupational Health, Ksaverska cesta 2, 10000 Zagreb, Croatia
b Biochemistry and Organic Analytical Chemistry Unit, Institute for Medical Research and Occupational Health, Ksaverska cesta 2, 10000 Zagreb, Croatia
c Department of Environmental Health, Dr. Andrija Štampar Institute of Public Health, Mirogojska cesta 16, 10000 Zagreb, Croatia
Abstract:Despite that the use of DDT has been restricted for more than 40 years to malaria affected areas, low doses of this pesticide and its metabolites DDE and DDD can be found in the environment around the world. Although it has been shown that these pollutants induce cell and DNA damage, the mechanisms of their cytogenotoxic activity remains largely unknown. This study looks into their possible genotoxic effects, at doses that can be found in body fluids, on human lymphocytes using the cytokinesis-block micronucleus assay and the comet assay. After exposure for 1, 6, and 24 h compounds p,p′-DDT (0.1 μg mL−1), p,p′-DDE (4.1 μg mL−1), and p,p′-DDD (3.9 μg mL−1) showed increase in DNA damage. The most significant results were observed at exposure period of 24 h where number of micronucleated cells increased from control 2.5 ± 0.71 to 23.5 ± 3.54, 13.5 ± 0.71, and 16.5 ± 6.36 for DDT, DDE, and DDD, respectively. Similar effect was observed using comet test where the percentage of DNA in comets tail increased from control 1.81 ± 0.16 to 17.24 ± 0.55, 11.21 ± 0.56 and 9.28 ± 0.50 for each compound, respectively. At the same time Fpg-comet assay failed to report induction of oxidative DNA damage of these pollutants. Additionally, the type of cell death was determined using diffusion assay and necrosis dominated. Our findings suggest that even at low concentrations, these pesticides could induce cytogenetic damage to human peripheral blood lymphocytes and in that manner have the impact on human health as well.
Keywords:CBPI  cytokinesis-block proliferation index  DDD  1  1-dichloro-2  2-bis(p-chlorophenyl)ethane  DDE  1  1-dichloro-2  2-bis(p-chlorophenyl)ethylene  DDT  1  1  1-trichloro-2  2-bis(p-chlorophenyl) ethane  DEET  N  N-diethyl-meta-toluamide  Fpg  formamidopyrimidine-DNA glycosylase  HR  high resolution  LMP  low melting point  NMP  normal melting point  MN  micronucleus  NB  nuclear bud  NPB  nucleoplasmic bridge
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