The “systems biology” in the study of xenobiotic effects.
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The project consist in a field monitoring program to evaluate the effects of toxic contaminants (e.g. heavy metals and hydrocarbons) on “sea sentinels” (organisms representing early living warning systems) purposely placed in different contaminated areas.
In our laboratory, we analyze by quali/quantitative PCR and ultrastructure analysis, tissues coming from Mytilus galloprovincialis placed in a polluted or unpolluted natural marine environment. Specifically, target tissues (gills, and digestive gland) are extracted to isolate RNA which is retrotranscribed to evaluated the presence of transcripts of the genes involved in oxidative stress, heavy metal toxicity and apoptosis, such as metallothionein (MT), heat shock protein 70 (HSP70), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), caspase-3, p53). The fquantitative evaluation of the expression of these key genes, together with Transmission Electron Microscopy (TEM) analysis, help to understand the type of environment pollution (presence of metallic nanoparticles), and adaptive responses triggered by these animal models. Such information is essential to give the basis for innovative interventions based on the use of in situ bioremediation using zerovalent iron nanoparticle system. Extremely promising, nanotechnology play a pivotal role also in environmental remediation, especially in respect of persistent organic (i.e. chlorophenols) and inorganic (i.e. Cr VI) pollutants.
For this research, we take advantage of several basic molecular biology techniques such as:
- DNA and RNA extraction
- DNA and RNA electrophoresis
- Qualitative and Real Time PCR
- cDNA cloning