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

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

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

STUDIES IN THE AGAVACEAE. I. CHROMOSOME MORPHOLOGY AND NUMBER OF SEVEN SPECIES

Volume: 21Start Page: 208End Page: 22

Genotoksični učinci karbamatnog insekticida Pirimor-50® na vršni meristem korijena biljke Vicia faba i kulturu ljudskih limfocita nakon izravne primjene i tretiranja njegovim metaboličkim ekstraktima

The aim of the study was to evaluate genotoxic effects of Pirimor-50®, a pirimicarb-based formulation (50 % active ingredient), in human lymphocyte cultures and Vicia faba root meristems. Furthermore, the objective was to examine a combined influence of insecticide treatment with mammalian microsomal S9 and vegetal S10 metabolic fractions or S10 mix metabolic transformation extracts (after Vicia faba primary roots treatment with Pirimor-50®). We used sister chromatid exchange assay-SCE and measured cell cycle progression and proliferation (proportion of M1-M3 metaphases and replication index ratio-RI). Two processes were used for plant promutagen activation: in vivo activation-Pirimor-50® was applied for 4 h to the plant and then S10 mix was added to lymphocytes; and, in vitro activation-lymphocytes were treated with Pirimor-50® and S10 or S9 for 2 h. Direct treatment induced significantly higher SCE frequencies in meristems at 0.01 mg mL-1. In lymphocytes, significantly higher SCE was at 1 mg mL-1 with decrease in RI and M1-M3 metaphase proportions at 0.5 mg mL-1 and cell division stop at 2.5 mg mL1. S10 mix lymphocyte treatment showed significantly elevated SCE values at 2-2.5 mg mL-1, with cell death at 3 mg mL-1. Lymphocyte treatment with Pirimor-50® together with S9 or S10 showed slightly elevated SCE frequency but had a significant influence on RI decrease, with lowest values in S9 treatment. Since no data are available on the genotoxicity of Pirimor-50®, this study is one of the first to evaluate and compare its direct effect in two bioassays, animal and vegetal, and also the effect of plant and animal metabolism on its genotoxic potential.Cilj ovog ispitivanja bio je procijeniti genotoksične učinke insekticida Pirimor-50®, formulacije koja se temelji na pirimikarbu (50 %-tni aktivni sastojak), u kulturama ljudskih limfocita i meristemu korijena biljke Vicia faba. Nadalje, cilj je bio ispitati objedinjeni utjecaj tretiranja insekticidom i metaboličkom mješavinom S9 za kultivirane stanice sisavaca i mješavinom S10 za stanice biljaka ili mješavinom S10 transformacijskih ekstrakata (nakon tretiranja primarnih korijena biljke Vicia faba insekticidom Pirimor-50®). Korišten je test izmjena sestrinskih kromatida (Sister Chromatid Exchange – SCE) i mjerena je progresija i proliferacija staničnog ciklusa (kroz omjer M1-M3 metafaza i vrijednost replikacijskog indeksa – RI). Dva su procesa korištena za aktivaciju biljnog promutagena: in vivo aktivacija – Pirimor-50® primjenjivan je tijekom 4 h na biljci, a potom je mješavina S10 dodana limfocitima, i in vitro aktivacija – limfociti su 2 h tretirani insekticidom Pirimor-50® i mješavinom S10 ili S9. Izravno tretiranje proizvelo je značajno veću učestalost SCE-a u meristemu pri 0,01 mg mL-1. U limfocitima je razina SCE-a značajno povećana pri 1 mg mL-1, uz smanjenje RI-ja i omjera M1-M3 metafaza pri 0,5 mg mL-1, uz zastoj stanične diobe pri 2,5 mg mL-1. Tretiranje limfocita mješavinom S10 značajno je povisilo vrijednosti SCE-a pri 2-2,5 mg mL-1, a stanična smrt nastupila je pri 3 mg mL-1. Tretiranje limfocita insekticidom Pirimor-50®, zajedno sa S9 ili S10, pokazalo je nešto veću učestalost SCE-a, ali i značajniji utjecaj na povećanje RI-ja, pri čemu su najniže vrijednosti utvrđene nakon tretiranja mješavinom S9. S obzirom na to da nema podataka o genotoksičnosti insekticida Pirimor-50®, ovo je istraživanje među prvima koje pomoću dvaju testova, životinjskoga i biljnoga, ispituje i uspoređuje njegov izravan učinak, ali i učinak biljnog i životinjskog metabolizma na njegov genotoksični potencijal

CONCENTRACIÓN TOTAL Y GEODISPONIBLE DE ELEMENTOS POTENCIALMENTE TÓXICOS EN SUELOS VOLCÁNICOS CON USO AGRÍCOLA DEL NEVADO DE TOLUCA, MÉXICO

En el Nevado de Toluca, México el uso indiscriminado de agroquímicos para la pro - ducción de alimentos ha generado impactos negativos en el suelo, como cambios en sus propiedades físicas, químicas y bioquímicas, lo que representa un riesgo potencial para la salud humana y el ambiente. El objetivo del presente estudio fue evaluar la concentración total y geodisponible de los elementos potencialmente tóxicos (As, Pb, Cd, Zn, Cu y Fe) en suelos agrícolas del Nevado de Toluca en los que se practica agri - cultura intensiva, semi-intensiva y tradicional. La evaluación de la concentración de los elementos potencialmente tóxicos se realizó por espectroscopia de emisión atómica por plasma de acoplamiento inductivo. En el caso del As, por espectroscopia de absorción atómica con generación de hidruros. Los resultados indican que en los suelos en los que se practica agricultura intensiva y semi-intensiva existe una mayor concentración total (mg/kg) de elementos potencialmente tóxicos que en los suelos en los que se practica agricultura tradicional. En algunos elementos potencialmente tóxicos se observó un aumento evidente en su geodisponibilidad (mg/L), no obstante que las concentraciones en términos absolutos son bajas. El uso no controlado de agroquímicos es un factor de degradación de los suelos agrícolas del Nevado de Toluca ya que modifica sus propie - dades físicas y químicas, principalmente en los que se practica la agricultura intensiva y semi-intensiva para el cultivo de papa. La adición de agroquímicos ha causado una fuerte acidificación (pH = 2.8 - 4.2) y consecuente intemperismo acelerado de las arcillas, que ha favorecido la reducción de la capacidad de intercambio catiónico, la disminución de la materia orgánica y el aumento de la conductividad eléctrica. Por tanto, los resultados de este estudio evidencian el riesgo ambiental (calidad de suelos) y a la salud humana en el largo plazo por el abuso de agroquímicos

Concentración total y geodisponible de elementos potencialmente tóxicos en suelos volcánicos con uso agrícola del nevado de Toluca, México

En el Nevado de Toluca, México el uso indiscriminado de agroquímicos para la pro - ducción de alimentos ha generado impactos negativos en el suelo, como cambios en sus propiedades físicas, químicas y bioquímicas, lo que representa un riesgo potencial para la salud humana y el ambiente. El objetivo del presente estudio fue evaluar la concentración total y geodisponible de los elementos potencialmente tóxicos (As, Pb, Cd, Zn, Cu y Fe) en suelos agrícolas del Nevado de Toluca en los que se practica agri - cultura intensiva, semi-intensiva y tradicional. La evaluación de la concentración de los elementos potencialmente tóxicos se realizó por espectroscopia de emisión atómica por plasma de acoplamiento inductivo. En el caso del As, por espectroscopia de absorción atómica con generación de hidruros. Los resultados indican que en los suelos en los que se practica agricultura intensiva y semi-intensiva existe una mayor concentración total (mg/kg) de elementos potencialmente tóxicos que en los suelos en los que se practica agricultura tradicional. En algunos elementos potencialmente tóxicos se observó un aumento evidente en su geodisponibilidad (mg/L), no obstante que las concentraciones en términos absolutos son bajas. El uso no controlado de agroquímicos es un factor de degradación de los suelos agrícolas del Nevado de Toluca ya que modifica sus propie - dades físicas y químicas, principalmente en los que se practica la agricultura intensiva y semi-intensiva para el cultivo de papa. La adición de agroquímicos ha causado una fuerte acidificación (pH = 2.8 - 4.2) y consecuente intemperismo acelerado de las arcillas, que ha favorecido la reducción de la capacidad de intercambio catiónico, la disminución de la materia orgánica y el aumento de la conductividad eléctrica. Por tanto, los resultados de este estudio evidencian el riesgo ambiental (calidad de suelos) y a la salud humana en el largo plazo por el abuso de agroquímicos