Paraoxon and glyphosate induce DNA double-strand breaks but are not type II topoisomerase poisons.

We tested the hypothesis that the pesticides paraoxon and glyphosate cause DNA double-strand breaks (DSB) by poisoning the enzyme Type II topoisomerase (topo II). Peripheral lymphocytes in G0 phase, treated with the pesticides, plus or minus ICRF-187, an inhibitor of Topo II, were stimulated to proliferate; induced cytogenetic damage was measured. Micronuclei, chromatin buds, nucleoplasmic bridges, and extranuclear fragments were induced by treatments with the pesticides, irrespective of the pre-treatment with ICRF-187. These results indicate that the pesticides do not act as topo II poisons. The induction of DSB may occur by other mechanisms, such as effects on other proteins involved in recombination repair.

Increased micronucleus frequencies in reticulocytes of children exposed to industrial pollution: oxidative stress and the OGG1 S326C polymorphism.

We examined possible early-effect biomarkers and polymorphisms of susceptibility in primary school children living near the Atoyac River in central México, which receives waste from multiple industries. We observed a significant increase in micronucleated reticulocytes associated with the oxidative stress index (OSI) and the OGG1 GG (S326C) genotype, and a significant decrease of reticulocytes carrying the transferrin receptor, inversely correlated with OSI.

Prenatal exposure to particulate matter and ozone: Bulky DNA adducts, plasma isoprostanes, allele risk variants, and neonate susceptibility in the Mexico City Metropolitan Area.

Mexico City's Metropolitan Area (MCMA) includes Mexico City and 60 municipalities of the neighbor states. Inhabitants are exposed to emissions from over five million vehicles and stationary sources of air pollutants such as particulate matter (PM) and ozone. MCMA PM contains elemental carbon and organic carbon (OC). OCs include polycyclic aromatic hydrocarbons (PAHs), many of which induce mutagenic and carcinogenic DNA adducts. Gestational exposure to air pollution has been associated with increased risk of intrauterine growth restriction, preterm birth or low birth weight risk, and PAH-DNA adducts. These effects also depend on the presence of risk alleles. We investigated the presence of bulky PAH-DNA adducts, plasma 8-iso-PGF2α (8-iso-prostaglandin F2α ) and risk allele variants in neonates cord blood and their non-smoking mothers' leucocytes from families that were living in a highly polluted area during 2014-2015. The presence of adducts was significantly associated with both PM2.5 and PM10 levels, mainly during the last trimester of gestation in both neonates and mothers, while the last month of pregnancy was significant for the association between ozone levels and maternal plasma 8-iso-PGF2α . Fetal CYP1B1*3 risk allele was associated with increased adduct levels in neonates while the presence of the maternal allele significantly reduced the levels of fetal adducts. Maternal NQO1*2 was associated with lower maternal levels of adducts. Our findings suggest the need to reduce actual PM limits in MCMA. We did not observe a clear association between PM and/or adduct levels and neonate weight, length, body mass index, Apgar or Capurro score. Environ. Mol. Mutagen. 60:428-442, 2019. © 2019 Wiley Periodicals, Inc.

Preclinical evidences of safety of a new synthetic adjuvant to formulate with the influenza human vaccine: absence of subchronic toxicity and mutagenicity.

Context: Influenza is a severe, life-threatening viral disease that can be prevented by vaccination. However, the anti-influenza human vaccine failed to show the required efficacy both in infants under 5 years old and in the elder population, who are among those with the highest risk of developing severe complications after influenza infection. Therefore, it is of high importance to improve the vaccine efficacy and ensure its safety in these susceptible populations. GK-1, a novel 18-aa peptide adjuvant, has been proved to increase the immunogenicity of the human influenza vaccine in both young and aged mice. Objective: A preclinical study of the toxicity profile of GK-1 following the World Health Organization guidelines to support its use was herein conducted. Material and methods: GK-1 was synthetically produced following Good Manufacturing Practices. The toxicological evaluation of GK-1 peptide was performed in rats after repeated dose-ranging trials by the subcutaneous route. The mutagenic potential of GK-1 was assessed by the micronucleus, chromosomal aberration, and Ames tests, in accordance with OECD Guidelines. Results: GK-1 did not show toxic effects at doses up to 12.5mg/kg, corresponding to 25 times the dose intended for human use. No indications of mutagenic potential were observed. GK-1 after dermal administration was well tolerated locally. Conclusion: The efficacy of GK-1 to improve influenza vaccine protection, along with the absence of toxicity and mutagenicity, as reported herein, support the evaluation of this peptide in a clinical trial as a novel adjuvant for human use.

Oxidative stress index is increased in children exposed to industrial discharges and is inversely correlated with metabolite excretion of voc.

Although the Atoyac River has been classified as highly polluted by environmental authorities, several communities are settled on its banks, affecting around 1.5 million persons, as well as farmland, due to an environmental distribution of toxics in the area. Our aim was to demonstrate that this environment affects important physiological processes that have an impact in health, so we conducted a study of schoolchildren from small communities on the banks of the river and in another similar town located far from it. 91 and 93 students, boys and girls, were studied from each site for oxidative stress index (OSI), calculated from the total antioxidant capacity and the total oxidative status, BTEX metabolite excretion and relevant metabolic polymorphisms participating in the bioactivation-detoxification of most VOC: CYP2E1 RsaI, NQO1 C609T, and null polymorphisms of GSTT1 and GSTM1. Results showed that OSI was significantly higher in children living by the river (5.23 ± 3.4 vs 2.59 ± 1.46, 95% C.I.). At this site, OSI was correlated with diminished metabolite excretion and a diminished antioxidant capacity; an association with genotypes CYP2E1RsaI (c2c2), GSTT1 present and NQO1*2 (CC) was also observed. Furthermore, boys at this site exhibited a diminished BMI compared to boys from the other community who were younger. IN CONCLUSION: children living at polluted sites like this, show early biological effects that might lead to health problems in their adult life. Environmental protection should be enforced to protect people's health in these sites where not even environmental monitoring is done. Environ. Mol. Mutagen. 59:639-652, 2018. © 2018 Wiley Periodicals, Inc.

Volatile Organic Compounds in Air: Sources, Distribution, Exposure and Associated Illnesses in Children.

BACKGROUND: Toxic volatile organic compounds (VOC), like benzene, toluene, ethylbenzene and xylenes (BTEX), are atmospheric pollutants representing a threat to human health. They are released into the environment from mobile sources in urban settings, but newly polluted areas are gaining importance in countries where accelerated industrialization is taking place in suburban or rural settings. METHODS: The review includes studies done in Mexico and Latin-America and countries considered to have emerging economies and are compared with similar studies in developed countries. Data about environmental VOC levels and exposure of children have been included. Also, information about health effects was reviewed. Articles were searched in PubMed and Scopus, and information was also obtained from the United States Environmental Protection Agency (EPA), the EPAs Integrated Risk Information System (IRIS-EPA) and state reports on air quality of Mexican cities. RESULTS: VOC or BTEX levels reported in industrial and suburban areas were found to be higher due to the burning of fossil fuels and waste emission; whereas, in big cities, VOC emissions were mainly due to mobile sources. Even though TEX levels were under reference values, benzene was found at levels several times over this value in cities and even higher in industrial zones. Elevated VOC emissions were also reported in cities with industrial development in their peripheral rural areas.Public health relevance: Industrial activities have changed the way of life of small towns, which previously had no concern about environmental pollution and chemicals. No air monitoring is done in these places where toxic chemicals are released into rivers and the atmosphere. This work demonstrates the need for environmental monitors to protect human life in suburban and rural areas where industrial growth occurs without planning and ecological or health protection, compromising the health of new generations beginning in fetal development.

Screening of Pesticides with the Potential of Inducing DSB and Successive Recombinational Repair.

A study was realized to ascertain whether eight selected pesticides would induce double strand breaks (DSB) in lymphocyte cultures and whether this damage would induce greater levels of proteins Rad51 participating in homologous recombination or of p-Ku80 participating in nonhomologous end joining. Only five pesticides were found to induce DSB of which only glyphosate and paraoxon induced a significant increase of p-Ku80 protein, indicating that nonhomologous end joining recombinational DNA repair system would be activated. The type of gamma-H2AX foci observed was comparable to that induced by etoposide at similar concentrations. These results are of importance since these effects occurred at low concentrations in the micromolar range, in acute treatments to the cells. Effects over longer exposures in actual environmental settings are expected to produce cumulative damage if repeated events of recombination take place over time.

Micronuclei in bone marrow and liver in relation to hepatic metabolism and antioxidant response due to coexposure to chloroform, dichloromethane, and toluene in the rat model.

Genotoxicity in cells may occur in different ways, direct interaction, production of electrophilic metabolites, and secondary genotoxicity via oxidative stress. Chloroform, dichloromethane, and toluene are primarily metabolized in liver by CYP2E1, producing reactive electrophilic metabolites, and may also produce oxidative stress via the uncoupled CYP2E1 catalytic cycle. Additionally, GSTT1 also participates in dichloromethane activation. Despite the oxidative metabolism of these compounds and the production of oxidative adducts, their genotoxicity in the bone marrow micronucleus test is unclear. The objective of this work was to analyze whether the oxidative metabolism induced by the coexposure to these compounds would account for increased micronucleus frequency. We used an approach including the analysis of phase I, phase II, and antioxidant enzymes, oxidative stress biomarkers, and micronuclei in bone marrow (MNPCE) and hepatocytes (MNHEP). Rats were administered different doses of an artificial mixture of CLF/DCM/TOL, under two regimes. After one administration MNPCE frequency increased in correlation with induced GSTT1 activity and no oxidative stress occurred. Conversely, after three-day treatments oxidative stress was observed, without genotoxicity. The effects observed indicate that MNPCE by the coexposure to these VOCs could be increased via inducing the activity of metabolism enzymes.

Assessment of exposure to organochlorine pesticides and levels of DNA damage in mother-infant pairs of an agrarian community.

Exposure to organochlorine pesticides was studied in a group of mother-infant pairs living in a rural area where agriculture is the main economic activity. Fumigation in this zone is performed with airplanes, thus affecting the inhabited areas around them, including schools. Heparinized venous blood of mothers and umbilical cords was used to evaluate the olive tail moment in the comet assay, and micronuclei, chromatin buds, and nucleoplasmic bridges in peripheral blood lymphocytes. Cord blood samples were taken at the moment of birth only from natural and normal parturitions. Determinations of hexachlorobenzene, aldrin, heptachlor epoxide, oxichlordane, t and c-chlordane, cis-nonachlor, mirex, alpha and beta-endosulfan, alpha, beta and gamma hexachlorocyclohexane, and p'p'-DDT, p'p'-DDE were conducted to establish the differential distribution of the toxicants between compartments, i.e., mother and umbilical cord. Significantly higher pesticide levels were found in umbilical cord plasma than in mothers' plasma for almost all compounds tested, except DDE and oxychlordane. Significantly higher olive tail moments were found in umbilical cords than in mothers, whereas micronuclei frequencies were higher in mothers than in umbilical cords. However, neither the levels of micronuclei nor the olive tail moment were correlated with pesticide levels. Given that no other exposure to toxic compounds has been identified in this region, the lack of correlation between genotoxicity biomarkers and pesticide levels may be due to the variability of the exposure and to endogenous processes related to lipid mobility during pregnancy, the metabolism of the compounds, and individual susceptibilities.

The HUman MicroNucleus project on eXfoLiated buccal cells (HUMN(XL)): the role of life-style, host factors, occupational exposures, health status, and assay protocol.

The human buccal micronucleus cytome assay (BMCyt) is one of the most widely used techniques to measure genetic damage in human population studies. Reducing protocol variability, assessing the role of confounders, and estimating a range of reference values are research priorities that will be addressed by the HUMN(XL) collaborative study. The HUMN(XL) project evaluates the impact of host factors, occupation, life-style, disease status, and protocol features on the occurrence of MN in exfoliated buccal cells. In addition, the study will provide a range of reference values for all cytome endpoints. A database of 5424 subjects with buccal MN values obtained from 30 laboratories worldwide was compiled and analyzed to investigate the influence of several conditions affecting MN frequency. Random effects models were mostly used to investigate MN predictors. The estimated spontaneous MN frequency was 0.74‰ (95% CI 0.52-1.05). Only staining among technical features influenced MN frequency, with an abnormal increase for non-DNA-specific stains. No effect of gender was evident, while the trend for age was highly significant (p<0.001). Most occupational exposures and a diagnosis of cancer significantly increased MN and other endpoints frequencies. MN frequency increased in heavy smoking (≥40cig/day, FR=1.37; 95% CI 1.03-.82) and decreased with daily fruit consumption (FR=0.68; 95% CI 0.50-0.91). The results of the HUMN(XL) project identified priorities for validation studies, increased the basic knowledge of the assay, and contributed to the creation of a laboratory network which in perspective may allow the evaluation of disease risk associated with MN frequency.

Genotoxic properties of nylon-6/MWNTs nanohybrid.

We performed the micronucleus test to determine the level of biocompatibility of pristine multi-walled carbon nanotubes (MWNTs) and MWNTs functionalized with nylon-6, (referred to as the nylon-6/ MWNTs nanohybrid), when they interact with human lymphocytes in a cell culture medium. A comparative genotoxic analysis demonstrated a better degree of biocompatibility of nylon-6/MWNT with human lymphocytes, as compared to pristine MWNTs, for the concentration range of 10-60 microg/ml. An evidence was found that pristine MWNTs act as clastogenic agents and possibly as aneuploidogenic agents, increasing the frequency of genotoxic bioindicators. On the other hand, nylon-6/ MWNTs nanohybrid were observed to induce cell death via apoptosis, which could be attributed to residual impurities of epsilon-caprolactam.