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Chemical structure of glyphosate, its metabolites?aminomethylphosphonic acid (AMPA), methylphosphonic acid and impurities?N-(phosphonomethyl)iminodiacetic acid (PMIDA), N-methylglyphosate, hydroxymethylphosphonic acid and bis-(phosphonomethyl)amine. doi:10.1371/journal.pone.0156946.g001
Source publication
The toxicity of herbicides to animals and human is an issue of worldwide concern. The present study has been undertaken to assess toxic effect of widely used pesticide - glyphosate, its metabolites: aminomethylphosphonic acid (AMPA) and methylphosphonic acid and its impurities: N-(phosphonomethyl)iminodiacetic acid (PMIDA), N-methylglyphosate, hydr...
Context in source publication
Context 1
... this study, hydroxymethylphosphonic acid and bis-(phosphonomethyl)amine have also been examined as potential impurities of glyphosate [13]. We have analyzed the effect of low concentrations (0.01 mM) of the xenobiotics studied on human PBMCs (environmental exposure), as well as high concentrations of these substances, which may enter human organism only as a result of acute poisoning (> 0.05 mM) (Fig 1). ...
Citations
... Moreover, nine independent association results were obtained in a large GWAS summary count of NK cell-related immune traits, including NK cells and CD45, SSC-A, CD16-CD56 on NK cells, CD45 on HLA-DR NK cells, CD8 + NKT cells, HLA DR NK/NK + cells, NK cells/CD3-lymphocytes, and HLA DR + NK cells/ CD3-lymphocytes. "SSC-A" is one of the parameters used to assess cell granularity in flow cytometry [23]. In the case of NK cells, it is their granularity that is measured, which allows NK cells to be easily distinguished from other immune cells [24]. ...
Background
Hepatocellular carcinoma (HCC), which is featured with high morbidity and mortality worldwide, is a primary malignant tumor of the liver. Recently, there is a wealth of supporting evidence revealing that NK cell-related immune traits are strongly associated with the development of HCC, but the causality between them has not been proven.
Methods
Two-sample Mendelian randomization (MR) study was performed to probe the causal correlation between NK cell-related immune traits and HCC. Genetic variations in NK cell-related immune traits were extracted from recent genome-wide association studies (GWAS) of individuals with European blood lineage. HCC data were derived from the UK Biobank Consortium's GWAS summary count data, including a total of 372,184 female and male subjects, with 168 cases and 372,016 controls, all of whom are of European ancestry. Sensitivity analysis was mainly used for heterogeneity and pleiotropy testing.
Results
Our research indicated the causality between NK cell-related immune traits and HCC. Importantly, CD8⁺NKT cells had protective causal effects on HCC (OR = 0.9996;95%CI,0.9993–0.9999; P = 0.0489). CD16⁻CD56 caused similar effects on NK cells (OR = 0.9997;95%CI,0.9996–0.9999; P = 0.0117) as CD8⁺NKT cells. Intercepts from Egger showed no pleiotropy and confounding factors. Furthermore, insufficient evidence was found to support the existence of heterogeneity by Cochran's Q test.
Conclusion
MR analysis suggested that low CD8⁺NKT cells and CD16⁻CD56 expression on NK cells were linked with a higher risk of HCC.
... In this study, we used glyphosate, one of the most widely applied pesticides on the global market, 28 as a modeling example. Glyphosate has a number of metabolites reaching the environment, 29 and its primary metabolite AMPA [29][30][31][32][33][34] has been shown to be toxic to humans. 35,36 In addition, glyphosate and its metabolites are frequently detected in soil and crops, 34 and the active metabolite (i.e., AMPA) is occasionally found at higher levels than glyphosate in plant tissues. ...
... In this study, we used glyphosate, one of the most widely applied pesticides on the global market, 28 as a modeling example. Glyphosate has a number of metabolites reaching the environment, 29 and its primary metabolite AMPA [29][30][31][32][33][34] has been shown to be toxic to humans. 35,36 In addition, glyphosate and its metabolites are frequently detected in soil and crops, 34 and the active metabolite (i.e., AMPA) is occasionally found at higher levels than glyphosate in plant tissues. ...
Although several models of pesticide uptake into plants are available, there are few modeling studies on the bioconcentration of metabolites in plants. Ignoring metabolites in plant uptake models can result in an underestimation of the parent compound's overall impacts on human health associated with pesticide residues in harvested food crops. To address this limitation, we offer a metabolite-based plant uptake model to predict the bioconcentration of the parent compound and its metabolites in plants. We used the uptake of glyphosate and its major metabolite (aminomethylphosphonic acid, AMPA) into potato as an example. The analysis of variability revealed that soil properties (affecting the soil sorption coefficient), dissipation half-life in soil, and metabolic half-life in the potato had a significant impact on the simulated AMPA concentration in the potato, indicating that regional variability could be generated in the plant bioconcentration process of metabolites. The proposed model was further compared using the non-metabolite model. The findings of the comparison suggested that the non-metabolite model, which is integrated with the AMPA bioconcentration process, can predict the AMPA concentration in the potato similarly to the proposed model. In conclusion, we provide insight into the bioconcentration process of metabolites in tuber plants from a modeling viewpoint, with some crucial model inputs, such as biotransformation and metabolic rate constants, requiring confirmation in future studies. The modeling demonstration emphasizes that it is relevant to consider bioaccumulation of metabolites, which can propagate further into increased overall residues of harmful compounds, especially in cases where metabolites have higher toxicity effect potency than their respective parent compounds.
... In general, it is recognized that commercial glyphosate-based herbicides cause higher cytotoxicity than pure glyphosate. 22,48,49 These findings might be due to a considerable toxicity of surfactants and the presence of amine salts of glyphosate. 50 Additional evidence that the cytotoxicity of commercial formulations containing glyphosate is higher than technical-grade glyphosate is supported by Townsend et al. 28 In this study, human Raji cells were exposed to concentrations of 17 μg/L-2,550 mg/L of technical-grade glyphosate (95% purity) for 24 h. ...
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.
... Moreover, toxic effects of exposure to glyphosate have also been observed in mammals and humans and the nervous system of mice [14]. Recent studies have reported its endocrine-disrupting activity in humans [15][16][17][18]. Kwiatkowska et al. [19] found that exposure to glyphosate can induce DNA damage in human leukocytes, resulting in epigenetic alterations in animal/human cells. ...
Glyphosate is a broad-spectrum herbicide widely employed in agriculture. Exposure to this genotoxic and endocrine-disrupting compound has adverse effects on terrestrial and aquatic organisms and on humans as well. Here, we explored the effects of glyphosate on female reproductive output and somatic growth rate in the marine polychaete worm, Ophryotrocha diadema. Adult focal individuals were exposed to different concentrations of pure glyphosate (0.0, 0.125 0.250, 0.500, 1.000 µg/mL) administered once a week for 3 weeks. Toxic effects and mortalities were observed at the three higher concentrations, whereas only a decrease in growth rate was noted after exposure to 0.125 µg/mL, which did not affect female allocation. An area of focus in future studies should be the effects of contaminants, their metabolites, and ecologically relevant human-driven stressors in the context of global warming.
... Glyphosate cytotoxicity and genotoxicity have been demonstrated across several cell types; however, whether these results are environmentally relevant remains debated. For example, Kwiatkowska et al. (2016) showed that glyphosate isopropylamine salt affects the viability of human peripheral blood mononuclear (PBMC) cells at 10 mM (tested 0.01-10 mM; 24-h exposure; propidum ioidide test). Similarly, in human fibroblast and fibrosarcoma cell lines, concentrations >5.2 and >4 mM, respectively, decreased cell viability (tested 4-6.5 mM; 4-h exposure; trypan blue staining; Monroy et al. 2005). ...
Glyphosate is the most applied agricultural chemical worldwide and has become nearly ubiquitous throughout the environment. Glyphosate is an effective herbicide because it disrupts the shikimate pathway, which is responsible for the synthesis of essential amino acids in plants and microorganisms. Given that there is no known target for glyphosate in higher animals, its toxicity to humans and other animals is heavily debated, especially after the 2015 IARC ruling that glyphosate is carcinogenic. Today, a growing body of literature shows in vitro, in vivo, and epidemiological evidence for the toxicity of glyphosate across animal species. With the application of glyphosate increasing globally, it is important to discuss these reports to enable a broader conversation on glyphosate toxicity and its impact on human and environmental health. Here, we summarize the recent glyphosate literature and discuss its implications.
... The MPA anion, which contains the phosphonic acid group and is a known metabolite of GPS, interacted with II in a comparable manner ( Figure 19) displaying 1:1 stoichiometry. 149 The association constants of MPA-TBA and MPA-THA with II were determined by fitting of absorption and fluorescence spectra and are shown in Table 3 Table 4 summarizes the bathochromic shifts observed for the prepolymerization mixtures in chloroform. 14 (18) 18 (17) MIP particles (with GPS-TBA and GPS-THA) and dNIP particles (with MPA-TBA and MPA-THA) were therefore synthesized in chloroform, yielding MIPTBA@SiO2 MIPTHA@SiO2, dNIPTBA@SiO2 and dNIPTHA@SiO2 ...
In dieser Arbeit wurden fluoreszierende MIP-Partikel hergestellt, um polare Analyten von biologischer und ökologischer Bedeutung zu analysieren, zum Beispiel den Krebs-Biomarker Sialinsäure (SA) und die Herbizide Glyphosat (GPS), Haloxyfop P und Fenoxaprop P. Die MIP-Schale wurde auf Siliziumdioxid und siliziumdioxid-beschichtete Polystyrol (PS)-Kernpartikel synthetisiert und enthielt einen Fluoreszenzfarbstoff, dessen Signal in Anwesenheit der Zielanalyten verstärkt wurde. Die Leistungsfähigkeit aller MIP-Partikel wurde durch den Vergleich ihres Bindungsverhaltens mit den nicht geprägten Polymer (NIP)-Partikeln und/oder durch ihre Diskriminierungsfähigkeit gegenüber konkurrierenden Analyten bewertet. Fluoreszierende Monomer für die MIP-Synthese erwiesen sich als vorteilhaft, da sowohl die Bindungsleistung als auch die Assoziationskonstanten einfacher bestimmt werden konnten. Als Alternative zur traditionellen Wasserstoffbrückenbindung zwischen Analyt und Bindungseinheit des Farbstoffes in unpolaren Lösungsmitteln, kann die Deprotonierung-Protonierung von harnstoffhaltigen Farbstoffen als Sensing-Mechanismus in polaren Medien wie DMF verwendet werden. Darüber hinaus führte die Verwendung eines „Dummy“-Analyten für die NIP-Partikel-Synthese zu einer Polymerschicht mit ähnlichen Eigenschaften wie die der MIP-Partikel, womit "Dummy"-NIPs als deutlich bessere Kontrollpartikel herangezogen werden konnten. Insgesamt erwies sich die Bestimmung der Diskriminierung der MIPs gegenüber konkurrierenden Analyten als konsistenteres Maß für die Leistungsbeurteilung. Mit Siliziumdioxid beschichtete, farbstoffdotierte PS-Partikel erwiesen sich als vielseitige Plattform für das Aufwachsen fluoreszierender MIP-Schalen, um zweifach fluoreszierende MIPs zu erhalten. Das Vorhandensein eines Kontrollsignals ist vorteilhaft für die Korrektur instrumenteller Schwankungen während analytischer Tests.
... In 2015, the EFSA declared that there is no straightforward evidence sustaining cytotoxicity and genotoxicity of Glyphosate, thus in vitro studies on different type of cells are required to better clarify the issue ((EFSA), 2015). Since then, a few studies showed that Glyphosate can be cytotoxic in several types of cells, and other in vitro adverse effects were described (Kwiatkowska et al., 2016;Kwiatkowska et al., 2017;Martínez et al., 2007;Nagy et al., 2019;Santovito et al., 2018). However, no information is available on thyroid cells. ...
... After adherence, we treated FRTL-5 cells with Glyphosate at increasing concentrations 0; 0,1; 0,25; 0,5; 1; 2; 10 mM, which derived from previous studies performed on other cell types. (Kwiatkowska et al., 2016;Kwiatkowska et al., 2017;Martínez et al., 2020;Nagy et al., 2019). Moreover, we took into account that acceptable daily intake (ADI) of 0.5 mg/kg was considered by EFSA corresponding to 0.500 μg/mL (3 mM) (EFSA, 2015;Santovito et al., 2018). ...
... It is important to observe that the effect of Glyphosate on cell viability was not dose-dependent. This is in line with previous findings in other cell types, in which both the cytotoxic and anti-proliferative effect were not dose-dependent (De Almeida et al., 2018;Kwiatkowska et al., 2016;Townsend et al., 2017). However, the fact that cytotoxicity was observed using two different assay techniques (WST-1 and Trypan blue) and both in the 2-D and the 3-D models, strongly support the concept that Glyphosate does exert a cytotoxic effect on thyroid cells, although it appears as a non-monotonic one. ...
Glyphosate is a pesticide, which contaminates the environment and exposes workers and general population to its residues present in foods and waters. In soil, Glyphosate is degraded in metabolites, amino-methyl-phosphonic acid (AMPA) being the main one. Glyphosate is considered a potential cancerogenic and endocrine-disruptor agent, however its adverse effects on the thyroid were evaluated only in animal models and in vitro data are still lacking. Aim of this study was to investigate whether exposure to Glyphosate could exert adverse effects on thyroid cells in vitro.
Two models (adherent-2D and spheroid-3D) derived from the same cell strain Fisher-rat-thyroid-cell line-5 (FRTL-5) were employed. After exposure to Glyphosate at increasing concentrations (0.0, 0.1–0.25- 0.5-1.0-2.0–10.0 mM) we evaluated cell viability by WST-1 (adherent and spheroids), results being confirmed by propidium-iodide staining (only for spheroids). Proliferation of adherent cells was assessed by crystal violet and trypan-blue assays, the increasing volume of spheroids was taken as a measure of proliferation. We also evaluated the ability of cells to form spheroids after Glyphosate exposure. We assessed changes of reactive-oxygen-species (ROS) by the cell-permeant H2DCFDA. Glyphosate-induced changes of mRNAs encoding for thyroid-related genes (TSHR, TPO, TG, NIS, TTF-1 and PAX8) were evaluated by RT-PCR.
Glyphosate reduced cell viability and proliferation in both models, even if at different concentrations. Glyphosate at the highest concentration reduced the ability of FRTL-5 to form spheroids. An increased ROS production was found in both models after exposure to Glyphosate. Finally, Glyphosate increased the mRNA levels of some thyroid related genes (TSHR, TPO, TG and TTF-1) in both models, while it increased the mRNAs of PAX8 and NIS only in the adherent model.
The present study supports an adverse effect of Glyphosate on cultured thyroid cells. Glyphosate reduced cell viability and proliferation and increased ROS production in thyroid cells.
... GLY at 0.036 g/L increased intracellular Ca 2+ concentrations in rat Sertoli cells [46] due to its ability to enhance mitochondrial membrane permeability, which finally leads to decreased ATP synthesis [47]. Inhibition of ATP synthesis following GLY exposure was observed also in human peripheral blood mononuclear cells, although in the millimolar range [48], and in swine granulosa cells in the micromolar range [49]. In female adult mice administered 250 or 500 mg/kg bw GLY, ATP decrease was observed in the ovary, with a concurrent increase in ROS levels and decrease of the mitochondrial membrane potential [50]. ...
Humans are exposed to residues of organophosphate and neonicotinoid pesticides, commonly used in agriculture. Children are particularly vulnerable and, among possible adverse outcomes, the increased incidence of premature mammary gland development (thelarche) has raised concern. We evaluated the toxicological effects of chlorpyrifos (CPF), imidacloprid (IMI) and glyphosate (GLY) at exposure concentrations occurring in children on the tumorigenic MCF-7 and non-tumorigenic MCF-12A breast cell lines, as representative of the target organ model, assessing cytotoxicity, apoptosis, necrosis, intracellular reactive oxygen species (ROS) and ATP levels, 17β-estradiol secretion and gene expression of nuclear receptors involved in mammary gland development. The pesticides decreased cell vitality in MCF-7 and cell proliferation in MCF-12A cells. ATP levels were decreased in MCF-7 cells by pesticides and apoptosis was increased in MCF-12A cells only by GLY (2.3 nM). ROS production was decreased by pesticides in both cell lines, except IMI (1.6 nM) in MCF-7 cells. Endocrine disrupting activity was highlighted by induction of 17β-estradiol secretion and modulation of the gene expression of estrogen alpha and beta, progesterone, androgen, and aryl hydrocarbon receptors in both cell lines. The use of MCF-7 and MCF-12A cells highlighted dissimilar modes of action of each pesticide at low human relevant concentrations.
... For example, the surface soil glyphosate concentrations were approximately 16% and 3% of the initial concentrations one and two weeks after the application, respectively, and at the end of the application period (i.e., 30 days), the glyphosate concentration was near zero. The AMPA concentration started to increase after the application of glyphosate and reached a Note: a The MF P→M value is estimated by dividing the molecular weight of AMPA (111 g mol − 1 ) by the molecular weight of glyphosate (169 g mol − 1 ), assuming a 100% transformation to simplify the simulation, because AMPA is a primary degradation product of glyphosate (Kwiatkowska et al., 2016;Roberts et al., 1998). ...
As pesticides can be degraded to toxic metabolites in the soil, metabolite toxicity should be considered in human health risk assessments. In this study, a screening-level modeling framework was developed to manage pesticides in surface soil, which was discussed under discrete and continuous emission scenarios. In addition, we selected glyphosate and its major metabolite (aminomethylphosphonic acid or AMPA) as examples to conduct screening-level risk management at regional, national, and global scales. The results indicated that if soil AMPA were not considered, human health risks could be significantly underestimated because of the large half-life of AMPA in the soil. For example, the added concentration factors of AMPA were simulated as 0.19 and 6.72 considering all major elimination pathways and considering the degradation pathway alone, respectively, indicating that AMPA formation could lead to severe extra health burdens. Furthermore, the evaluation of current glyphosate soil standards suggested that toxic metabolites should be considered in the regulatory process; otherwise, many standards could theoretically trigger high levels of soil AMPA, which could result in serious human health damage. Our proposed screening-level model can help to improve risk assessment and regulatory management of pesticides in surface soils.
... Pesticide TPs may be more toxic and persistent than the initial pesticide [119]. For instance, more pronounced negative effects in comparison with the parent compound are shown for the main bioactive metabolites of glyphosateaminomethylphosphonic acid (AMPA) and N-(phosphonomethyl)iminodiacetic acid (PMIDA) [40,120]; carbosulfancarbofuran and 3-hydroxycarbofuran [121] and imidacloprid [18]. Moreover, the impact can be implicitly expressed. ...
Pesticides are commonly used in agriculture to enhance crop production and control pests. Therefore, pesticide residues can persist in the environment and agricultural crops. Although modern formulations are relatively safe to non-target species, numerous theoretical and experimental data demonstrate that pesticide residues can produce long-term negative effects on the health of humans and animals and stability of ecosystems. Of particular interest are molecular mechanisms that mediate the start of a cascade of adverse effects. This is a review of the latest literature data on the effects and consequences of contamination of agricultural crops by pesticide residues. In addition, we address the issue of implicit risks associated with pesticide formulations. The effects of pesticides are considered in the context of the Adverse Outcome Pathway concept.
