Search

Polycyclic aromatic hydrocarbons (PAHs) are known to exert mutagenic and carcinogenic effects. Research on extracted organic matter (EOM) from environmental mixtures has indicated several mechanisms of intracellular damage in living organisms. The toxic effect of environmental pollutants is usually assessed on cell systems or in single species. We used the model organism Caenorhabditis elegans to compare the effect of synthetic PAHs with that of the EOM from environmental mixtures. The biological effect was measured by monitoring the expression level of some crucial genes, sensitive parameters of the organism's response. The results indicate the ability of C. elegans to counteract damage by mounting a stress-response only in the presence of EOM. On the other hand the exposure of C. elegans to a mixture of synthetic PAHs determines the silencing of the transcriptional machinery, thus preventing the synthesis of proteins that are important for both the damage repair mechanism and survival itself. The results strongly indicate that the study of environmental toxicant effects at the molecular level may provide information on their mechanism of action. Copyright © 2011 John Wiley & Sons, Ltd.

Background FAD synthase is a ubiquitous enzyme that catalyses the last step of FAD biosynthesis, allowing for the biogenesis of several flavoproteins. In humans different isoforms are generated by alternative splicing, isoform 1 being localized in mitochondria. Homology searching in Caenorabditis elegans leads to the identification of two human FAD synthase homologues, coded by the single copy gene R53.1 . Methods The C. elegans R53.1 gene was silenced by feeding. The expression level of transcripts was established by semi-quantitative RT-PCR. Overall protein composition was evaluated by two-dimensional electrophoresis. Enzymatic activities were measured by spectrophotometry and oxygen consumption by polarography on isolated mitochondria. Results From R53.1 two transcripts are generated by trans -splicing. Reducing by 50% the transcription efficiency of R53.1 by RNAi results in a 50% reduction in total flavin with decrease in ATP content and increase in ROS level. Significant phenotypical changes are noticed in knock-down nematodes. Among them, a significant impairment in locomotion behaviour possibly due to altered cholinergic transmission. At biochemical level, impairment of flavoenzyme activities and of some KCN-insensitive oxygen-consuming enzymes is detected. At proteomic level, at least 15 abundant proteins are affected by R53.1 gene silencing, among which superoxide dismutases. Conclusion and General Significance For the first time we addressed the existence of different isoforms of FAD-metabolizing enzymes in nematodes. A correlation between FAD synthase silencing and flavoenzyme derangement, energy shortage and redox balance impairment is apparent. In this aspect R53.1 -interfered nematodes could provide an animal model system for studying human pathologies with alteration in flavin homeostasis/flavoenzyme biogenesis.

Le recenti politiche europee sono rivolte alla valutazione dei possibili effetti nocivi di miscele complesse di sostanze chimiche (REACH) utilizzando metodi che escludano l'impiego di vertebrati. In questa relazione vengono illustrati gli effetti di tossicità di particolato atmosferico su due organismi modello: Caenorhabditis elegans e Rhodobacter sphaeroides. I campioni di particolato sono stati raccolti in due siti della città di Bari e da questi sono stati estratti i composti organici solubili in acetone ed esano. I risultati ottenuti da tali test hanno mostrato un effetto tossico sul nematode C. elegans, mentre nel caso del batterio fotosintetico gli effetti sono limitati ad una leggera diminuzione della velocità di crescita. Gli sviluppi futuri di questo studio saranno orientati verso frazioni granulometriche quali PM1 e nano particolato e verso altri sistemi modello.

Polycyclic aromatic hydrocarbons (PAHs) are known to exert mutagenic and carcinogenic effects. Research on extracted organic matter (EOM) from environmental mixtures has indicated several mechanisms of intracellular damage in living organisms. The toxic effect of environmental pollutants is usually assessed on cell systems or in single species. We used the model organism Caenorhabditis elegans to compare the effect of synthetic PAHs with that of the EOM from environmental mixtures. The biological effect was measured by monitoring the expression level of some crucial genes, sensitive parameters of the organism's response. The results indicate the ability of C. elegans to counteract damage by mounting a stress-response only in the presence of EOM. On the other hand the exposure of C. elegans to a mixture of synthetic PAHs determines the silencing of the transcriptional machinery, thus preventing the synthesis of proteins that are important for both the damage repair mechanism and survival itself. The results strongly indicate that the study of environmental toxicant effects at the molecular level may provide information on their mechanism of action.