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Assessment of DNA damage by RAPD in Paracentrotus lividus embryos exposed to amniotic fluid from residents living close to waste landfill sites.

Guida M, Guida M, De Felice B, Santafede D, D'Alessandro R, Di Spiezio Sardo A, Scognamiglio M, Ferrara C, Bifulco G, Nappi C - J. Biomed. Biotechnol. (2010)

Bottom Line: The aim of this study was to assess the genotoxic effects of environmental chemicals on residents living near landfills.The study was based on samples of amniotic fluid from women living in the intensely polluted areas around the Campania region of Italy compared to a nonexposed control group.These results show the ability of RAPD-PCR to detect and isolate DNA sequences representing genetic alterations induced in P. lividus embryos.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetric Gynecology Urological Sciences and Reproductive Medicine, University of Naples Federico II, Via Pansini, Napoli, Italy.

ABSTRACT
The aim of this study was to assess the genotoxic effects of environmental chemicals on residents living near landfills. The study was based on samples of amniotic fluid from women living in the intensely polluted areas around the Campania region of Italy compared to a nonexposed control group. We evaluated the genetic effects that this amniotic fluids collected in contaminated sites had on Paracentrotus lividus embryos. DNA damage was detected through changes in RAPD (Random Amplified Polymorphism DNA) profiles. The absence of the amplified DNA fragments indicated deletions in Paracentrotus lividus DNA exposed to the contaminated amniotic fluids when compared to equal exposure to uncontaminated fluids. These results show the ability of RAPD-PCR to detect and isolate DNA sequences representing genetic alterations induced in P. lividus embryos. Using this method, we identified two candidate target regions for DNA alterations in the genome of P. lividus. Our research indicates that RAPD-PCR in P. lividus embryo DNA can provide a molecular approach for studying DNA damage from pollutants that can impact human health. To our knowledge, this is the first time that assessment of DNA damage in P. lividus embryos has been tested using the RAPD strategy after exposure to amniotic fluid from residents near waste landfill sites.

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Comparison of developmental defects and mortality in P. lividus embryos following exposure over the range of zygote to pluteus stage.  The gray bar represents the untreated negative control (C0). Unpolluted and polluted amniotic liquid-induced developmental defects are represented, respectively, by white and black bars. P1, per cent larval malformations; P2, per cent developmental arrest at blastula/gastrula stage; D, per cent embryonic mortality, N, normal larvae. Results are expressed as means ± SE from  six separate experiments.
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fig1: Comparison of developmental defects and mortality in P. lividus embryos following exposure over the range of zygote to pluteus stage. The gray bar represents the untreated negative control (C0). Unpolluted and polluted amniotic liquid-induced developmental defects are represented, respectively, by white and black bars. P1, per cent larval malformations; P2, per cent developmental arrest at blastula/gastrula stage; D, per cent embryonic mortality, N, normal larvae. Results are expressed as means ± SE from six separate experiments.

Mentions: The mean percentage of embryotoxicity (including all 15 samples run in six replicates) is shown in Figure 1. The developmental defects in polluted amniotic liquid-treated P. lividus larvae are mainly of the P1 type (larvae affected in skeletal or gut differentiation). In a lower number of embryos, the effect becomes drastic, with total arrest at prelarval stadium (P2). In particular, in polluted amniotic fluid samples, we have found the maximum quantity of malformed and dead larvae, which indicates the most severe toxicity class.


Assessment of DNA damage by RAPD in Paracentrotus lividus embryos exposed to amniotic fluid from residents living close to waste landfill sites.

Guida M, Guida M, De Felice B, Santafede D, D'Alessandro R, Di Spiezio Sardo A, Scognamiglio M, Ferrara C, Bifulco G, Nappi C - J. Biomed. Biotechnol. (2010)

Comparison of developmental defects and mortality in P. lividus embryos following exposure over the range of zygote to pluteus stage.  The gray bar represents the untreated negative control (C0). Unpolluted and polluted amniotic liquid-induced developmental defects are represented, respectively, by white and black bars. P1, per cent larval malformations; P2, per cent developmental arrest at blastula/gastrula stage; D, per cent embryonic mortality, N, normal larvae. Results are expressed as means ± SE from  six separate experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2913803&req=5

fig1: Comparison of developmental defects and mortality in P. lividus embryos following exposure over the range of zygote to pluteus stage. The gray bar represents the untreated negative control (C0). Unpolluted and polluted amniotic liquid-induced developmental defects are represented, respectively, by white and black bars. P1, per cent larval malformations; P2, per cent developmental arrest at blastula/gastrula stage; D, per cent embryonic mortality, N, normal larvae. Results are expressed as means ± SE from six separate experiments.
Mentions: The mean percentage of embryotoxicity (including all 15 samples run in six replicates) is shown in Figure 1. The developmental defects in polluted amniotic liquid-treated P. lividus larvae are mainly of the P1 type (larvae affected in skeletal or gut differentiation). In a lower number of embryos, the effect becomes drastic, with total arrest at prelarval stadium (P2). In particular, in polluted amniotic fluid samples, we have found the maximum quantity of malformed and dead larvae, which indicates the most severe toxicity class.

Bottom Line: The aim of this study was to assess the genotoxic effects of environmental chemicals on residents living near landfills.The study was based on samples of amniotic fluid from women living in the intensely polluted areas around the Campania region of Italy compared to a nonexposed control group.These results show the ability of RAPD-PCR to detect and isolate DNA sequences representing genetic alterations induced in P. lividus embryos.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetric Gynecology Urological Sciences and Reproductive Medicine, University of Naples Federico II, Via Pansini, Napoli, Italy.

ABSTRACT
The aim of this study was to assess the genotoxic effects of environmental chemicals on residents living near landfills. The study was based on samples of amniotic fluid from women living in the intensely polluted areas around the Campania region of Italy compared to a nonexposed control group. We evaluated the genetic effects that this amniotic fluids collected in contaminated sites had on Paracentrotus lividus embryos. DNA damage was detected through changes in RAPD (Random Amplified Polymorphism DNA) profiles. The absence of the amplified DNA fragments indicated deletions in Paracentrotus lividus DNA exposed to the contaminated amniotic fluids when compared to equal exposure to uncontaminated fluids. These results show the ability of RAPD-PCR to detect and isolate DNA sequences representing genetic alterations induced in P. lividus embryos. Using this method, we identified two candidate target regions for DNA alterations in the genome of P. lividus. Our research indicates that RAPD-PCR in P. lividus embryo DNA can provide a molecular approach for studying DNA damage from pollutants that can impact human health. To our knowledge, this is the first time that assessment of DNA damage in P. lividus embryos has been tested using the RAPD strategy after exposure to amniotic fluid from residents near waste landfill sites.

Show MeSH
Related in: MedlinePlus