Toxicity assessment and DNA repair kinetics in HEK293 cells exposed to environmentally relevant concentrations of Glyphosate (Roundup® Control Max).

Glyphosate is a systemic, non-selective, pre and post-emergence wide range herbicide. In 2015, IARC classified Glyphosate as "a probable carcinogenic agent for humans". The aim of this study was to evaluate the cytotoxicity and genotoxicity of the commercial formulation of glyphosate (Roundup® Control Max) at environmentally relevant concentrations and measure the potential effect of this herbicide over the cell capacity to repair DNA damage. HEK293 cells were exposed to 5 concentrations of Roundup® Control Max equivalent to 0.7; 7; 70; 700 and 3,500 µg/L glyphosate acid, for 1, 4 and 24 h. Cytotoxicity was quantified by the Trypan Blue staining method and by the MTT assay, while genotoxicity and evaluation of DNA damage repair kinetics were analyzed through the alkaline comet assay. In all treatments, cell viability was higher than 80%. The three highest glyphosate concentrations-70 µg/L, 700 µg/L, and 3,500 µg/L-increased levels of DNA damage compared to the control at the three exposure times tested. Finally, concerning the kinetics of DNA damage repair, cells initially exposed to 3,500 µg/L of glyphosate for 24 h were unable to repair the breaks in DNA strands even after 4 h of incubation in culture medium. The present study demonstrated for the first time that Roundup® Control Max may induce genetic damage and cause alterations in the DNA repair system in human embryonic kidney cells even at concentrations found in blood and breast milk of people exposed through residues of the herbicide in food, which values have been poorly assessed or not studied yet according to the existent literature.

Cytogenetic markers and their spatial distribution in a population living in proximity to areas sprayed with pesticides.

Human populations are in contact with potentially toxic substances in varying amounts, if the exposure is work-related or direct, generally the amount of toxin is usually greater than if the exposure is environmental through the drifts that occur. It was proposed to determine the existence of genotoxic damage evaluated through Chromosomal Aberrations and Micronuclei assays and their spatial distribution pattern, as well as the possible relationship between that damage and the values found in biochemical biomarkers, in groups of individuals environmental exposure (respiratory exposure) to mixtures of pesticides, in the province of Córdoba-Argentina. Biochemical and hematological determinations were made in each samples. The results reveal that the monitoring of human populations through the analysis of cytogenetic markers enabled the detection of direct damage in man caused by polluting substances and the results were obtained rapidly. The disadvantage of this type of study is the inability to estimate the degree of exposure.

Evaluation of genetic damage in pesticides applicators from the province of Córdoba, Argentina.

The impact evaluation of pesticide exposure is conducted using combined data from biomonitoring and environmental monitoring. Damage to the human genome is, probably, the leading cause of chronic-degenerative disorders, reproductive toxicology, and developmental problems. Although the general population is exposed to pesticides, workers in the agrochemical industry and farmers represent a high-risk group due to the occupational and environmental exposure. The aim of this study is to determine whether occupational exposure to agrochemicals in Córdoba (Argentina) constitute a factor of genotoxic damage. The study was conducted in 30 pesticide applicators from the province of Córdoba. Chromosomal aberrations (CAs), micronuclei (MN), and comet assays (CO) were performed. The current study shows that occupational exposure to pesticides increases values of CAs, MN, and DNA fragmentation biomarkers, all indicators of damage to the genetic material. Evidence suggests that chronic exposure to pesticides is a potential risk to workers health.

Evaluación del nivel de daño en el material genético de niños de la provincia de Córdoba expuestos a plaguicidas / Assessment of the level of damage to the genetic material of children exposed to pesticides in the province of Córdoba

Introducción. En las últimas décadas, numerosos autores han investigado acerca de los daños genotóxicos producidos por la exposición a sustancias químicas, aunque no existen para Argentina reportes de estudios que analicen dichos efectos en los niños. El objetivo de este trabajo fue determinar la frecuencia de micronúcleos en células exfoliadas de la mucosa bucal en niños de plantas urbanas con exposición ambiental (por inhalación) y compararla con la frecuencia de micronúcleos en niños que habitan en plantas urbanas alejadas de zonas donde se pulveriza. Población y métodos. Se estudiaron cincuenta niños que habitan la localidad de Marcos Juárez (Córdoba), ubicados a diferentes distancias de exposición a la aplicación de productos plaguicidas, y veinticinco niños de la ciudad de Río Cuarto (Córdoba), considerados no expuestos a dichos productos, y se aplicó el ensayo de micronúcleos en células de la mucosa bucal. Resultados. Se encontró diferencia significativa entre los expuestos a menos de quinientos metros con respecto al grupo de niños no expuestos. El 40% de los individuos expuestos sufren algún tipo de afección persistente, que se podría asociar a la exposición crónica a plaguicidas. Conclusiones. Los resultados obtenidos permiten indicar que existe una exposición a sustancias genotóxicas en un grupo de niños con relación al otro y poner de manifiesto la relevancia del ensayo de micronúcleos en la mucosa bucal para el biomonitoreo genético y la vigilancia en salud pública. El ensayo utilizado detecta un nivel de daño que todavía es reversible.

Introduction. In the past decades, several authors have investigated the genotoxicity caused by exposure to chemicals, but there are no reports on studies analyzing such effects on children in Argentina. The objective of this study was to establish the micronucleus frequency in exfoliated buccal mucosa cells in children from urban areas with environmental exposure (through inhalation) and to compare it with the micronucleus frequency in children from urban regions far from areas subjected to spraying. Population and Methods. Fifty children living in the town of Marcos Juárez (Córdoba) at different distances from pesticide spraying areas and twenty-five children from the city of Río Cuarto (Córdoba), who are considered not exposed to pesticides, were studied; the micronucleus assay in buccal mucosa cells was used. Results. A significant difference was observed between exposed children living less than 500 m from areas subjected to spraying and those who were not exposed. Forty percent of exposed children suffer some type of persistent condition, which may be associated with chronic exposure to pesticides. Conclusions. Results indicate that genotoxicity is present in a group of children compared to the other one, and highlight the importance of the micronucleus assay in buccal mucosa cells for genetic biomonitoring and public health surveillance. This assay is capable of detecting a level of damage that can be reversible.

Evaluación del nivel de daño en el material genético de niños de la provincia de Córdoba expuestos a plaguicidas / Assessment of the level of damage to the genetic material of children exposed to pesticides in the province of Córdoba

Introducción. En las últimas décadas, numerosos autores han investigado acerca de los daños genotóxicos producidos por la exposición a sustancias químicas, aunque no existen para Argentina reportes de estudios que analicen dichos efectos en los niños. El objetivo de este trabajo fue determinar la frecuencia de micronúcleos en células exfoliadas de la mucosa bucal en niños de plantas urbanas con exposición ambiental (por inhalación) y compararla con la frecuencia de micronúcleos en niños que habitan en plantas urbanas alejadas de zonas donde se pulveriza. Población y métodos. Se estudiaron cincuenta niños que habitan la localidad de Marcos Juárez (Córdoba), ubicados a diferentes distancias de exposición a la aplicación de productos plaguicidas, y veinticinco niños de la ciudad de Río Cuarto (Córdoba), considerados no expuestos a dichos productos, y se aplicó el ensayo de micronúcleos en células de la mucosa bucal. Resultados. Se encontró diferencia significativa entre los expuestos a menos de quinientos metros con respecto al grupo de niños no expuestos. El 40% de los individuos expuestos sufren algún tipo de afección persistente, que se podría asociar a la exposición crónica a plaguicidas. Conclusiones. Los resultados obtenidos permiten indicar que existe una exposición a sustancias genotóxicas en un grupo de niños con relación al otro y poner de manifiesto la relevancia del ensayo de micronúcleos en la mucosa bucal para el biomonitoreo genético y la vigilancia en salud pública. El ensayo utilizado detecta un nivel de daño que todavía es reversible.(AU)

Introduction. In the past decades, several authors have investigated the genotoxicity caused by exposure to chemicals, but there are no reports on studies analyzing such effects on children in Argentina. The objective of this study was to establish the micronucleus frequency in exfoliated buccal mucosa cells in children from urban areas with environmental exposure (through inhalation) and to compare it with the micronucleus frequency in children from urban regions far from areas subjected to spraying. Population and Methods. Fifty children living in the town of Marcos Juárez (Córdoba) at different distances from pesticide spraying areas and twenty-five children from the city of Río Cuarto (Córdoba), who are considered not exposed to pesticides, were studied; the micronucleus assay in buccal mucosa cells was used. Results. A significant difference was observed between exposed children living less than 500 m from areas subjected to spraying and those who were not exposed. Forty percent of exposed children suffer some type of persistent condition, which may be associated with chronic exposure to pesticides. Conclusions. Results indicate that genotoxicity is present in a group of children compared to the other one, and highlight the importance of the micronucleus assay in buccal mucosa cells for genetic biomonitoring and public health surveillance. This assay is capable of detecting a level of damage that can be reversible.(AU)

Assessment of the level of damage to the genetic material of children exposed to pesticides in the province of Córdoba.

INTRODUCTION: In the past decades, several authors have investigated the genotoxicity caused by exposure to chemicals, but there are no reports on studies analyzing such effects on children in Argentina. The objective of this study was to establish the micronucleus frequency in exfoliated buccal mucosa cells in children from urban areas with environmental exposure (through inhalation) and to compare it with the micronucleus frequency in children from urban regions far from areas subjected to spraying. POPULATION AND METHODS: Fifty children living in the town of Marcos Juárez (Córdoba) at different distances from pesticide spraying areas and twenty-five children from the city of Río Cuarto (Córdoba), who are considered not exposed to pesticides, were studied; the micronucleus assay in buccal mucosa cells was used. RESULTS: A significant difference was observed between exposed children living less than 500 m from areas subjected to spraying and those who were not exposed. Forty percent of exposed children suffer some type of persistent condition, which may be associated with chronic exposure to pesticides. CONCLUSIONS: Results indicate that genotoxicity is present in a group of children compared to the other one, and highlight the importance of the micronucleus assay in buccal mucosa cells for genetic biomonitoring and public health surveillance. This assay is capable of detecting a level of damage that can be reversible.

[Turner's syndrome 45,X/46Xdel(X)(q21): a case report]. / Síndrome de Turner con mosaicismo 45,X/46Xdel(X)(q21). Comunicación de un caso.

Turner's syndrome was described by Otto Ullrich (1930) and Henry Turner (1938). An estimated 1 from 2,000 to 3,000 female babies and 1% of the conceptions of female embryos and fetuses have this condition, and 95 to 99% of them result in miscarriage during the first trimester. The case presented concerns a 15 y/o girl who consulted due to primary amenorrhea. The karyotype was 45,X[6]/46Xdel(X)(q21)[14]. Her mother had experienced premature ovarian failure and her karyotype was: 46Xdel(X)(q21)[3]/46,XX[35].

Síndrome de Turner con mosaicismo 45,X/46Xdel(X)(q21): Comunicación de un caso / Turner's syndrome 45,X/46Xdel(X)(q21): A case report

El síndrome de Turner fue descrito por Otto Ullrich en 1930 y por Henry Turner en 1938. Se estima que 1 de 2000 a 3000 niñas recién nacidas, y 1% de las concepciones de embriones y fetos femeninos portan esta patología, llegándose a abortar espontáneamente, en el primer trimestre, entre el 95% y el 99% de los fetos afectados. El caso que se presenta corresponde a una adolescente, nacida en 1995, que consultó por amenorrea primaria, con cariotipo 45,X[6]/46Xdel(X)(q21)[14]. Dado el resultado observado, se estudió a la madre, quien había experimentado una insuficiencia ovárica prematura y cuyo cariotipo era 46Xdel(X)(q21) [3]/ 46,XX[35].

Turner's syndrome was described by Otto Ullrich (1930) and Henry Turner (1938). An estimated 1 from 2,000 to 3,000 female babies and 1% of the conceptions of female embryos and fetuses have this condition, and 95 to 99% of them result in miscarriage during the first trimester. The case presented concerns a 15 y/o girl who consulted due to primary amenorrhea. The karyotype was 45,X[6]/46Xdel(X)(q21)[14]. Her mother had experienced premature ovarian failure and her karyotype was: 46Xdel(X)(q21)[3]/46,XX[35].

Síndrome de Turner con mosaicismo 45,X/46Xdel(X)(q21): Comunicación de un caso / Turners syndrome 45,X/46Xdel(X)(q21): A case report

El síndrome de Turner fue descrito por Otto Ullrich en 1930 y por Henry Turner en 1938. Se estima que 1 de 2000 a 3000 niñas recién nacidas, y 1% de las concepciones de embriones y fetos femeninos portan esta patología, llegándose a abortar espontáneamente, en el primer trimestre, entre el 95% y el 99% de los fetos afectados. El caso que se presenta corresponde a una adolescente, nacida en 1995, que consultó por amenorrea primaria, con cariotipo 45,X[6]/46Xdel(X)(q21)[14]. Dado el resultado observado, se estudió a la madre, quien había experimentado una insuficiencia ovárica prematura y cuyo cariotipo era 46Xdel(X)(q21) [3]/ 46,XX[35].(AU)

Turners syndrome was described by Otto Ullrich (1930) and Henry Turner (1938). An estimated 1 from 2,000 to 3,000 female babies and 1% of the conceptions of female embryos and fetuses have this condition, and 95 to 99% of them result in miscarriage during the first trimester. The case presented concerns a 15 y/o girl who consulted due to primary amenorrhea. The karyotype was 45,X[6]/46Xdel(X)(q21)[14]. Her mother had experienced premature ovarian failure and her karyotype was: 46Xdel(X)(q21)[3]/46,XX[35].(AU)

Síndrome de Turner con mosaicismo 45,X/46Xdel(X)(q21). Comunicación de un caso. / [Turners syndrome 45,X/46Xdel(X)(q21): a case report].

Turners syndrome was described by Otto Ullrich (1930) and Henry Turner (1938). An estimated 1 from 2,000 to 3,000 female babies and 1

of the conceptions of female embryos and fetuses have this condition, and 95 to 99

of them result in miscarriage during the first trimester. The case presented concerns a 15 y/o girl who consulted due to primary amenorrhea. The karyotype was 45,X[6]/46Xdel(X)(q21)[14]. Her mother had experienced premature ovarian failure and her karyotype was: 46Xdel(X)(q21)[3]/46,XX[35].

Biomarcadores d daño genético en poblaciones humanas expuestas a plaguicidas / Biomarkers of Genetic Damage in Human Populations Exposed to Pesticides

El efecto de los plaguicidas sobre la salud humana, animal y ambiental es preocupación de la comunidad científica desde hace mucho tiempo. Numerosos estudios reportan que los plaguicidas no son inofensivos y que su uso puede conducir a efectos biológicos dañinos a mediano y a largo plazo, en los grupos humanos y animales expuestos, en el presente o en los descendientes. La importancia en la detección precoz del daño genético radica en que permite tomar las medidas necesarias para disminuir o suprimir la exposición al agente deletéreo cuando aún éste es reversible, y de ese modo prevenir y disminuir el riesgo de desarrollar neoplasias y otras alteraciones patológicas. En este trabajo se revisan los principales conceptos en la temática, la utilidad de los estudios de genotoxicidad y se hace referencia a los trabajos realizados en los últimos veinticinco años sobre monitoreo genético de personas expuestas laboralmente a plaguicidas. Los ensayos de genotoxicidad, que incluyen aberraciones cromosómicas, micronúcleos, intercambio de cromátidas hermanas y cometa, deberían ser considerados como herramientas indispensables en la implementación de una vigilancia médica completa en personas potencialmente expuestas a diversos contaminantes ambientales y en especial aquellas que habitan en el mismo lugar con personas que ya han desarrollado algún tipo de neoplasia en edades tempranas, con el fin de prevenir la ocurrencia de tumores de origen ambiental y especialmente laboral.

The effect of pesticides on human, animal and environmental health has been cause of concern in the scientific community for a long time. Numerous studies have reported that pesticides are not harmless and that their use can lead to harmful biological effects in the medium and long term, in exposed human and animals, and their offspring. The importance of early detection of genetic damage is that it allows us to take the necessary measures to reduce or eliminate the exposure to the deleterious agent when damage is still reversible, and thus to prevent and to diminish the risk of developing tumors or other alterations. In this paper we reviewed the main concepts in the field, the usefulness of genotoxicity studies and we compiled studies performed during the last twenty years on genetic monitoring of people occupationally exposed to pesticides. We think that genotoxicity tests, including that include chromosomal aberrations, micronucleous, sister chromatid exchanges and comet assays, should be considered as essential tools in the implementation of complete medical supervision for people exposed to potential environmental pollutants, particularly for those living in the same place as others who where others have already developed some type of malignancy. This action is particularly important at early stages to prevent the occurrence of tumors, especially from environmental origins.

Evaluación del daño en el material genético en grupos humanos expuestos a agroquímicos para la prevención de patologías asociadas / Assessment of genetic material damage in human groups exposed to agrochemicals for the prevention of related diseases

INTRODUCCIÓN El monitoreo de grupos de población humana expuestos a agentes potencialmente dañinos es una herramienta valiosa en salud pública y ocupacional. Tiene como propósito preservar la salud y la calidad de vida en aquellos grupos de población humana que son de alto riesgo debido a la naturaleza de las sustancias a las que están expuestos. OBJETIVOS Obtener elementos para proponer un protocolo de vigilancia de las medidas preventivas y de vigilancia de la salud de la familia rural, que sirva como aporte para acciones futuras en salud pública. Discutir la problemática de la exposición a tóxicos ambientales/ocupacionales como factores de riesgo en la etiología de enfermedades maligna. MÉTODOS El estudio se basó en un diseño transversal de tipo caso-control. Se realizaron encuestas con el fin de obtener información sobre actividad laboral y antecedentes personales. Para detectar en forma temprana los efectos biológicos de los plaguicidas antes de que causen daño en la salud, se emplearon los ensayos de aberraciones cromosómicas y micronúcleos (MN) en sangre y mucosa bucal. RESULTADOS Se observaron diferencias estadísticamente significativas entre los dos grupos estudiados en los tres ensayos y una asociación entre los ensayos de MN en sangre y mucosa bucal. DISCUSIÓN Los resultados permiten obtener elementos para proponer un protocolo de vigilancia que sirva como aporte para acciones futuras en salud pública. La evaluación de los efectos de los agroquímicos sobre el material genético de grupos humanos expuestos conlleva un deber de concientización sobre medidas de protección personal, control y seguridad ambiental.