Procjena genotoksičnosti insekticida Lannate-90® i njegovih biljnih i životinjskih metabolita u kulturi ljudskih limfocita

This study evaluated direct and metabolic genotoxic effects caused by Lannate-90®, a methomyl-based formulation (90 % active ingredient), in human lymphocyte cultures using sister chromatid exchange assay (SCE). Two processes were used for the plant promutagens evaluation: in vivo activation, applying the insecticide systemically in plants for 4 h and subsequently adding plant metabolites containing extracts to lymphocyte cultures; and in vitro activation, where the insecticide was incubated with Vicia faba S10 mix plus human lymphocyte culture. Direct treatment with the insecticide significantly increased SCE frequency in human lymphocytes (250-750 mg L-1), with cellular death observed at 1000 mg L-1 concentration. Using the extracts of Vicia faba treated with Lannate-90® to treat human lymphocytes, a dose-response relationship was observed. In lymphocyte cultures treated directly with the insecticide for 2 h, a negative response was obtained. When S10 mix was added, SCE frequency did not change significantly. Meanwhile, a mixture of S9 mammalian metabolic mix and Lannate-90® increased the SCE frequency, with an observed concentration-dependent response. Although Lannate-90® induced cellular death at the highest concentrations, it did not cause a delay in cell proliferation in any of the treatments, confirming its genotoxic action. This study is one of the first to evaluate and compare the direct effect of Lannate-90® in two bioassays, animal and vegetal, and the effect of plant and animal metabolism on its genotoxic potential.Korištenjem testa izmjena sestrinskih kromatida (eng. Sister Chromatide Exchange Assay – SCE) u kulturama ljudskih limfocita ispitivani su izravni i metabolički genotoksični učinci insekticida Lannate-90®, formulacije koja se temelji na metomilu (90 % aktivni sastojak). Za procjenu biljnih promutagena provedena su dva postupka: in vivo aktivacija, kod koje se insekticid četiri sata sustavno primjenjivao na biljci, a potom su kulturama limfocita dodani biljni metaboliti s ekstraktom, i aktivacija in vitro, kod koje je insekticid inkubiran mješavinom S10 biljke Vicia faba i kulturom ljudskih limfocita. Izravno tretiranje insekticidom značajno je povećalo učestalost SCE-a u ljudskim limfocitima (250-750 mg L-1), a stanična smrt uočena je pri koncentraciji od 1000 mg L-1. Nakon tretiranja ljudskih limfocita ekstraktima biljke Vicia faba koji su tretirani insekticidom Lannate-90®, primijećen je odnos između doze i učinka. Kod kultura limfocita koje su dva sata bile izravno tretirane insekticidom primijećen je negativan odgovor. Kada je dodana S10 mješavina za metaboličku aktivaciju, učestalost SCE-a nije se značajnije promijenila. Naspram tomu, metabolička mješavina S9 za kultivirane stanice sisavaca i Lannate-90® povećali su učestalost SCE-a, uz zamijećen koncentracijski ovisan odgovor. Premda je Lannate-90® inducirao staničnu smrt pri najvišim koncentracijama, nije uzrokovao zastoj stanične proliferacije ni u jednom postupku, čime se potvrđuje njegovo genotoksično djelovanje. Ovo je ispitivanje među prvima kojim se procjenjivao i uspoređivao izravan učinak insekticida Lannate-90® u dvama biološkim testovima, životinjskom i biljnom, te učinak biljnog i životinjskog metabolizma na njegov genotoksični potencijal

Evaluation of Genotoxic and Cytotoxic Effects in Human Peripheral Blood Lymphocytes Exposed In Vitro to Neonicotinoid Insecticides News

Calypso (thiacloprid), Poncho (clothianidin), Gaucho (imidacloprid), and Jade (imidacloprid) are commercial neonicotinoid insecticides, a new class of agrochemicals in México. However, genotoxic and cytotoxic studies have not been performed. In the present study, human peripheral blood lymphocytes (PBL) were exposed in vitro to different concentrations of the four insecticides. The genotoxic and cytotoxic effects were evaluated using the alkaline comet and trypan blue dye exclusion assays. DNA damage was evaluated using two genotoxicity parameters: tail length and comet frequency. Exposure to 9.5 × 10−6 to 5.7 × 10−5 M Jade; 2.8 × 10−4 to 1.7 × 10−3 M Gaucho; 0.6 × 10−1 to 1.4 × 10−1 M Calypso; 1.2 × 10−1 to 9.5 × 10−1 M Poncho for 2 h induced a significant increase DNA damage with a concentration-dependent relationship. Jade was the most genotoxic of the four insecticides studied. Cytotoxicity was observed in cells exposed to 18 × 10−3 M Jade, 2.0 × 10−3 M Gaucho, 2.0 × 10−1 M Calypso, 1.07 M Poncho, and cell death occurred at 30 × 10−3 M Jade, 3.3 × 10−3 M Gaucho, 2.8 × 10−1 M Calypso, and 1.42 M Poncho. This study provides the first report of genotoxic and cytotoxic effects in PBL following in vitro exposure to commercial neonicotinoid insecticides

Procjena genotoksičnosti insekticida Lannate-90® i njegovih biljnih i životinjskih metabolita u kulturi ljudskih limfocita

This study evaluated direct and metabolic genotoxic effects caused by Lannate-90®, a methomyl-based formulation (90 % active ingredient), in human lymphocyte cultures using sister chromatid exchange assay (SCE). Two processes were used for the plant promutagens evaluation: in vivo activation, applying the insecticide systemically in plants for 4 h and subsequently adding plant metabolites containing extracts to lymphocyte cultures; and in vitro activation, where the insecticide was incubated with Vicia faba S10 mix plus human lymphocyte culture. Direct treatment with the insecticide significantly increased SCE frequency in human lymphocytes (250-750 mg L-1), with cellular death observed at 1000 mg L-1 concentration. Using the extracts of Vicia faba treated with Lannate-90® to treat human lymphocytes, a dose-response relationship was observed. In lymphocyte cultures treated directly with the insecticide for 2 h, a negative response was obtained. When S10 mix was added, SCE frequency did not change significantly. Meanwhile, a mixture of S9 mammalian metabolic mix and Lannate-90® increased the SCE frequency, with an observed concentration-dependent response. Although Lannate-90® induced cellular death at the highest concentrations, it did not cause a delay in cell proliferation in any of the treatments, confirming its genotoxic action. This study is one of the first to evaluate and compare the direct effect of Lannate-90® in two bioassays, animal and vegetal, and the effect of plant and animal metabolism on its genotoxic potential.Korištenjem testa izmjena sestrinskih kromatida (eng. Sister Chromatide Exchange Assay – SCE) u kulturama ljudskih limfocita ispitivani su izravni i metabolički genotoksični učinci insekticida Lannate-90®, formulacije koja se temelji na metomilu (90 % aktivni sastojak). Za procjenu biljnih promutagena provedena su dva postupka: in vivo aktivacija, kod koje se insekticid četiri sata sustavno primjenjivao na biljci, a potom su kulturama limfocita dodani biljni metaboliti s ekstraktom, i aktivacija in vitro, kod koje je insekticid inkubiran mješavinom S10 biljke Vicia faba i kulturom ljudskih limfocita. Izravno tretiranje insekticidom značajno je povećalo učestalost SCE-a u ljudskim limfocitima (250-750 mg L-1), a stanična smrt uočena je pri koncentraciji od 1000 mg L-1. Nakon tretiranja ljudskih limfocita ekstraktima biljke Vicia faba koji su tretirani insekticidom Lannate-90®, primijećen je odnos između doze i učinka. Kod kultura limfocita koje su dva sata bile izravno tretirane insekticidom primijećen je negativan odgovor. Kada je dodana S10 mješavina za metaboličku aktivaciju, učestalost SCE-a nije se značajnije promijenila. Naspram tomu, metabolička mješavina S9 za kultivirane stanice sisavaca i Lannate-90® povećali su učestalost SCE-a, uz zamijećen koncentracijski ovisan odgovor. Premda je Lannate-90® inducirao staničnu smrt pri najvišim koncentracijama, nije uzrokovao zastoj stanične proliferacije ni u jednom postupku, čime se potvrđuje njegovo genotoksično djelovanje. Ovo je ispitivanje među prvima kojim se procjenjivao i uspoređivao izravan učinak insekticida Lannate-90® u dvama biološkim testovima, životinjskom i biljnom, te učinak biljnog i životinjskog metabolizma na njegov genotoksični potencijal

The microRNAs as potential biomarkers for predicting the onset of aflatoxin exposure in human beings: a review

The identification of aflatoxins as human carcinogens has stimulated extensive research efforts, which continue to the present, to assess potential health hazards resulting from contamination of the human food supply and to minimize exposure. The use of biomarkers that are mechanistically supported by toxicological studies will be important tools for identifying stages in the progression of development of the health effects of environmental agents. miRNAs are small non-coding mRNAs that regulate post-transcriptional gene expression. Also, they are molecular markers of cellular responses to various chemical agents. Growing evidence has demonstrated that environmental chemicals can induce changes in miRNA expression. miRNAs are good biomarkers because they are well defined, chemically uniform, restricted to a manageable number of species, and stable in cells and in the circulation. miRNAs have been used as serological markers of HCC and other tumors. The expression patterns of different miRNAs can distinguish among HCC-hepatitis viruses related, HCC cirrhosis-derivate, and HCC unrelated to either of them. The main objective of this review is to find unreported miRNAs in HCC related to other causes, so that they can be used as specific molecular biomarkers in populations exposed to aflatoxins and as early markers of exposure, damage/presence of HCC. Until today specific miRNAs as markers for aflatoxins-exposure and their reliability are currently lacking. Based on their elucidated mechanisms of action, potential miRNAs that could serve as possible markers of HCC by exposure to aflatoxins are miR-27a, miR-27b, miR-122, miR-148, miR-155, miR-192, miR-214, miR-221, miR-429, and miR-500. Future validation for all of these miRNAs will be needed to assess their prognostic significance and confirm their relationship with the induction of HCC due to aflatoxin exposure