The situation of chlorpyrifos in Mexico: a case study in environmental samples and aquatic organisms.

Chlorpyrifos (CPF) is one of the most commonly used organophosphate pesticides. Because CPF was described as a toxic compound without safe levels of exposure for children, certain countries in Latin America and the European Union have banned or restricted its use; however, in Mexico it is used very frequently. The aim of this study was to describe the current situation of CPF in Mexico, as well as its use, commercialization, and presence in soil, water, and aquatic organisms in an agricultural region of Mexico. Structured questionnaires were applied to pesticide retailers to determine the sales pattern of CPF (ethyl and methyl); in addition, monthly censuses were conducted with empty pesticide containers to assess the CPF pattern of use. Furthermore, samples of soil (48 samples), water (51 samples), and fish (31 samples) were collected, which were analyzed chromatographically. Descriptive statistics were performed. The results indicate that CPF was one of the most sold (3.82%) and employed OP (14.74%) during 2021. Only one soil sample was found above the CPF limit of quantification (LOQ); in contrast, all water samples had CPF levels above the LOQ (x̄ = 4614.2 ng/L of CPF). In the case of fish samples, 6.45% demonstrated the presence of methyl-CPF. In conclusion, the information obtained in this study indicates the need for constant monitoring in the area, since the presence of CPF in soil, water, and fish constitutes a threat to the health of wildlife and humans. Therefore, CPF should be banned in Mexico to avoid a serious neurocognitive health problem.

Temephos, an organophosphate larvicide for residential use: a review of its toxicity.

Temephos (O,O,O',O'-tetramethyl O,O'-thiodi-p-phenylene bis(phosphorothioate)) is a larvicide belonging to the family of organophosphate pesticides used for the control of different vectors of diseases, such as dengue, Zika, chikungunya, and dracunculiasis. The aim of this review was to discuss the available published information about temephos toxicokinetics and toxicity in mammals. Temephos is quickly absorbed in the gastrointestinal tract, distributed to all organs, and then it accumulates mainly in adipose tissue. It is metabolized by S-oxidation, oxidative desulfuration, and hydrolysis reactions, with the possible participation of cytochrome P450 (CYP). Temephos is mainly eliminated by feces, whereas some of its metabolites are eliminated by urine. The World Health Organization classifies it as class III: slightly dangerous with a NOAEL (no-observed adverse effect level) of 2.3 mg/kg/day for up to 90 days in rats, based on brain acetylcholinesterase (AChE) inhibition. A LOAEL (lowest observable adverse effect level) of 100 mg/kg/day for up to 44 days in rats was proposed based on cholinergic symptoms. However, some studies have shown that temephos causes toxic effects in mammals. The inhibition of the enzyme acetylcholinesterase (AChE) is one of its main demonstrated effects; however, this larvicide has also shown genotoxic effects and some adverse effects on male reproduction and fertility, as well as liver damage, even at low doses. We performed an extensive review through several databases of the literature about temephos toxicokinetics, and we recommend to revisit current assessment of temephos with the new available data.

Toxicokinetics of temephos after oral administration to adult male rats.

Temephos (Tem) is the larvicide of choice to control mosquito transmission of dengue, Zika, and chikungunya. The toxicokinetic and toxicological information of temephos is very limited. The aim of this work was to determine the toxicokinetics and dosimetry of temephos and its metabolites. Male Wistar rats were orally administered temephos (300 mg/kg) emulsified with saline solution and sacrificed over time after dosing. Temephos and its metabolites were analyzed in blood and tissues by high performance liquid chromatography-diode array detector. At least eleven metabolites were detected, including temephos-sulfoxide (Tem-SO), temephos-oxon (Tem-oxon), temephos-oxon-sulfoxide (Tem-oxon-SO), temephos-oxon-SO-monohydrolyzed (Tem-oxon-SO-OH), 4,4´-thiodiphenol, 4,4´-sulfinyldiphenol, and 4,4´-sulfonyldiphenol or bisphenol S (BPS). The mean blood concentrations of temephos were fitted to a one-compartment model for kinetic analysis. At 2 h, the peak was reached (t1/2 abs = 0.38 h), and only trace levels were detected at 36 h (t1/2 elim = 8.6 h). Temephos was detected in all tissues and preferentially accumulated in fat. Temephos-sulfone-monohydrolyzed (Tem-SO2-OH) blood levels remained constant until 36 h and gradually accumulated in the kidney. Tem-oxon was detected in the brain, liver, kidney, and fat. Clearance from the liver and kidney were 7.59 and 5.52 ml/min, respectively. These results indicate that temephos is well absorbed, extensively metabolized, widely distributed and preferentially stored in adipose tissue. It is biotransformed into reactive metabolites such as Tem-oxons, Tem-dioxons, and BPS. Tem-SO2-OH, the most abundant metabolite of temephos, could be used as an exposure biomarker for toxicokinetic modeling. These results could provide critical insight into the dosimetry and toxicity of temephos and its metabolites.

Methamidophos induces cytotoxicity and oxidative stress in human peripheral blood mononuclear cells.

Previous studies have shown that organophosphate pesticide (OP) exposure is associated with oxidative stress. Methamidophos (MET) is an OP widely used in agriculture, which is regarded as a highly toxic pesticide and it is a potent inhibitor of acetylcholinesterase. The aim of this study was to evaluate whether MET can induce oxidative stress at low concentrations in primary cultures of human peripheral blood mononuclear cells (PBMCs). PBMCs from healthy individuals were exposed to MET (0-80 mg/L) for 0-72 h. We performed the MTT and neutral-red assays to assess the cytotoxicity. As indicators of oxidative stress, the levels of reactive oxygen species (ROS) were assessed using flow cytometry, and the malondialdehyde (MDA) and reduced glutathione (GSH) levels were determined. MET decreased the viability of PBMCs in a dose-dependent manner. At concentrations of 3, 10, or 20 mg/L for 24 h, MET increased the ROS production significantly compared with the vehicle control. Similarly, MET increased the levels of MDA at the same concentrations that increased ROS (10 and 20 mg/L); however, no changes in GSH levels were observed. These results suggest that MET increased the generation of oxidative stress in PBMCs. © 2015 Wiley Periodicals, Inc. Environ Toxicol 32: 147-155, 2017.

Downregulation of human paraoxonase 1 (PON1) by organophosphate pesticides in HepG2 cells.

Paraoxonase 1 (PON1) is a calcium-dependent esterase synthesized primarily in the liver and secreted into the plasma where it is associated with high-density lipoproteins (HDL). PON1 hydrolyzes and detoxifies some toxic metabolites of organophosphorus compounds (OPs) such as methyl parathion and chlorpyrifos. Thus, PON1 activity and expression levels are important for determining susceptibility against OPs poisoning. Some studies have demonstrated that OPs can modulate gene expression through interactions with nuclear receptors. In this study, we evaluated the effects of methyl parathion and chlorpyrifos on the modulation of PON1 in Human Hepatocellular Carcinoma (HepG2) cells by real-time PCR, PON1 activity assay, and western blot. The results showed that the treatments with methyl parathion and chlorpyrifos decreased PON1 mRNA and immunoreactive protein and increased inflammatory cytokines in HepG2 cells. The effects of methyl parathion and chlorpyrifos on the downregulation of PON1 gene expression in HepG2 cells may provide evidence of OPs cytotoxicity related to oxidative stress and an inflammatory response. A decrease in the expression of the PON1 gene may increase the susceptibility to OPs intoxication and the risk of diseases related to inflammation and oxidative stress. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 490-500, 2017.

Oxidative stress and genetic damage among workers exposed primarily to organophosphate and pyrethroid pesticides.

The indiscriminate use of pesticides in agriculture and public health campaigns has been associated with an increase of oxidative stress and DNA damage, resulting in health outcomes. Some defense mechanisms against free radical-induced oxidative damage include the antioxidant enzyme systems. The aim of this study was to determine the levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and the relationship of antioxidant enzyme levels with DNA damage among sprayers (workers) occupationally exposed to pesticides. The determinations of MDA and antioxidant enzymes were performed spectrophotometrically. The genotoxic effects were evaluated using the comet assay. The results showed a marginally significant decrease in SOD and CAT activities in the high exposure group compared to the control group. For MDA, statistically significant differences were found among people working long term vs. those working temporarily (P = 0.02) as sprayers. In the moderate exposure group, a positive correlation was observed between MDA levels and GPx activity. In the high exposure group, a negative correlation was observed between GR and CAT activities, and between MDA levels and GPx activities. Furthermore, in the high exposure group, a positive correlation between DNA damage parameters and MDA levels was observed. The results suggest an important role of antioxidant enzymes for the protection of DNA damage caused by occupational exposure to pesticides.

[Initial surgical management of squamous carcinoma of the vulva]. / Manejo quirúrgico inicial del carcinoma epidermoide vulvar.

Vulvar cancer accounts for approximately 4% of gynecological malignancies. At the Instituto Nacional de Cancerologia in Mexico it occupies the fourth place. The purpose of this study is to assess the management of squamous carcinoma of the vulva with initial surgical treatment. It is a descriptive retrospective, observational study, from January 1, 2002 to December 31, 2012. Twenty-seven patients, clinical stages I, II, or III, initial surgical management, with at least one year of follow-up were included. In 51.85% a partial vulvectomy was performed and in 40.74% a wide excision; 66.66% underwent inguinofemoral dissection. Recurrence occurred in 25.91% of cases and the overall survival at 10 years was 63%. It is concluded that with invasion of up to 1 mm of lymph node, affection is 0%; with invasion of 1 mm and up to 5 mm this increases to 25%; an invasion of more than 5 mm implies up to 45%. Recurrence in our study was primarily distant, necessitating long-term monitoring with emphasis on symptoms to request imaging studies when suspected. Adjuvant therapy should be offered to patients with positive nodes, close or positive margins, and tumors larger than 4 cm.

Trends on deaths from acute pesticide poisoning in Mexico, 2000-2021.

OBJETIVE: To provide a comprehensive analysis of mortality trends from acute pesticide poisoning in Mexico from 2000 through 2021. METHODS: The governmental records of deaths from acute pesticide poisoning were used. The age-standardized years of life lost and aged-standardized mortality rates were estimated. Significant changes in trends of annual percentage change were identified using Joinpoint regression. RESULTS: Between 2000 and 2021, mortality was primarily observed in individuals aged 15 to 19 years. Males were the most affected. Self-inflicted pesticide poisoning was the primary registered reason for death. The age-standardized mortality rate from acute pesticide poisoning was reduced from 2012 to 2021 (APC: -4.4; p=0.003). CONCLUSION: This report is the first study about the mortality rate from acute pesticide poisoning in Mexico. The results provided evidence to consider in developing laws to prevent acute pesticide poisoning.

Morbidity of breast cancer and cervico-uterine cancer in women from the occidental region of Mexico.

BACKGROUND: The incidences of breast cancer (BC) and cervico-uterine cancer (CC) vary widely from country to country. In Mexico, BC mortality has doubled in the last 20 years to become the second leading cause of death for women aged 30 to 54 years. CC is the most common cause of death from neoplasia in women over 25 years old. In 2006, the state of Nayarit had one of the highest mortality rates for these types of cancers in Mexico. OBJECTIVE: To analyze and characterize the current demographics and morbidities associated with BC and CC in the state of Nayarit. MATERIAL AND METHODS: In this retrospective study, the clinical histories of patients who were diagnosed with BC or CC at the State Cancer Center from January 2006 to December 2010 were analyzed. RESULTS: A total of 406 patients with BC and 328 patients with CC were registered. The most common clinical stage for both cancer types was IIB. The municipalities of San Pedro Lagunillas and El Nayar presented the highest prevalences of BC and CC, respectively. CONCLUISION: Our results suggest that women living in poorer and more marginalized regions have a higher possibility of developing BC and CC. Because BC and CC are preventable and treatable in their early stages, demographic information from population records for these cancers is helpful in determining the incidence rates and patterns and improving decision-making processes.

ß-glucuronidase as a biomarker for assessing the exposure to anticholinergic pesticides: A meta-analysis.

INTRODUCTION: The human exposure to anticholinergic pesticides has been associated with the development of various diseases. Therefore, several biomarkers have been proposed for biomonitoring human exposure to anticholinergic pesticides. OBJECTIVE: This work evaluated the effect of human exposure to anticholinergic pesticides on ß-glucuronidase (GUSB) levels. METHODS: A systematic review was performed using PubMed, Web of Science, Scopus, and EBSCO databases up to December 2021. The statistical analysis employed standardized mean differences and meta-regression. And the trial sequential analysis was performed. RESULTS: Nine studies were included. A monotonic relationship was observed between poisoning severity and GUSB. Furthermore, BuChE levels were correlated with GUSB levels. CONCLUSIONS: The results indicated that GUSB levels could be used as a possible diagnosis biomarker in poisoning related to anticholinergic pesticide exposure. However, the use of GUSB to assess the chronic exposure to anticholinergic pesticides could be only performed in recent exposure (≈ 7 days after last exposure).

Hematological indices as indicators of inflammation induced by exposure to pesticides.

Pesticide toxicity, both acute and chronic, is a global public health concern. Pesticides are involved in abnormal inflammatory responses by interfering with the normal physiology and metabolic status of cells. In this regard, inflammatory indices aggregate index of systemic inflammation (AISI), monocyte-to-high-density lipoprotein ratio, monocyte-to-lymphocyte ratio (MLR), neutrophil-to-lymphocyte platelet ratio (NLPR), neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, systemic immune inflammation index, and systemic inflammation response index (SIRI) have been used as predictive markers of inflammatory status in several diseases and also in acute poisoning events. This study aimed to determine systemic inflammation indices and their relationship with pesticide exposure from urban sprayers in 302 individuals categorized into three groups (reference group and moderate and high exposure groups). The data suggest that the AISI, MLR, NLPR, and SIRI indices were significantly higher in the exposed groups compared with the reference group. In conclusion, this study proposes that inflammation indices warrant further attention in order to assess their value as early biomarkers of acute and chronic pesticide intoxication.

Methylation patterns of the CDKN2B and CDKN2A genes in an indigenous population exposed to pesticides.

The INK4-ARF locus includes the CDKN2B and CDKN2A genes and is functionally relevant in the regulation of both cell proliferation and senescence. Studies have reported modifications of DNA methylation in this locus by exposure to environmental contaminants including pesticides; however, until now, specific methylation profiles have not been reported in genetically conserved populations exposed to occupational pesticides. The aim of this study was to determine the methylation profiles of the CDKN2B and CDKN2A genes in a genetically conserved population exposed to pesticides. A cross-sectional and analytical study was carried out in 190 Huichol indigenous persons. Information related to pesticide exposure, diet and other variables were obtained through the use of a structured questionnaire. Blood and urine samples were collected for methylation test and dialkylphosphates (DAP) determination, respectively. DNA methylation was measured by the pyrosequencing of bisulfite-treated DNA and DAP concentrations by gas chromatography-tandem mass spectrometry (GC/MS). The most frequent metabolite in the population was dimethylthiophosphate. The farmer group presented a higher methylation percentage of CDKN2B than the non-farmer group, but no differences in CDKN2A were observed between groups. A positive correlation between methylation of CpG site 3 of CDKN2B and time working in the field was observed in the farmer group. An association between methylation percentage of CDKN2B and age was also observed in the non-farmer group. These results suggest that pesticide exposure and exposure time in Huichol indigenous individuals could modify the methylation pattern of the CDKN2B gene.

The Relationship between Cancer and Paraoxonase 1.

Extensive research has been carried out to understand and elucidate the mechanisms of paraoxonase 1 (PON1) in the development of diseases including cancer, cardiovascular diseases, neurological diseases, and inflammatory diseases. This review focuses on the relationship between PON1 and cancer. The data suggest that PON1, oxidative stress, chronic inflammation, and cancer are closely linked. Certainly, the gene expression of PON1 will remain challenging to study. Therefore, targeting PON1, redox-sensitive pathways, and transcription factors promise prevention and therapy in the development of several diseases, including cancer.

Environmental and socio-cultural impacts on global DNA methylation in the indigenous Huichol population of Nayarit, Mexico.

Alterations of global DNA methylation have been evaluated in several studies worldwide; however, Long Interspersed Nuclear Elements-1 (LINE-1) methylation in genetically conserved populations such as indigenous communities have not, to our knowledge, been reported. The aim of this study was to evaluate the relationship between LINE-1 methylation patterns and factors such as pesticide exposure and socio-cultural characteristics in the Indigenous Huichol Population of Nayarit, Mexico. A cross-sectional study was conducted in 140 Huichol indigenous individuals. A structured questionnaire was used to determine general and anthropometric characteristics, diet, harmful habits, and pesticide exposure. DNA methylation was determined by pyrosequencing of bisulfite-treated DNA. A lower level of LINE-1 methylation was found in the indigenous population when compared to a Mestizo population previously studied by our group. This difference might be due to the influence of the genetic admixture and differing dietary and lifestyle habits. The males in the indigenous population exhibited increased LINE-1 methylation in comparison to the females. Sex and alcohol consumption showed positive associations with LINE-1 methylation, while weight, current work in the field, current pesticide usage, and folate intake exhibited negative associations with LINE-1 methylation. The results suggest that ethnicity, as well as other internal and environmental factors, might influence LINE-1 methylation.

Organophosphorus pesticide exposure biomarkers in a Mexican population.

The organophosphate (OP) pesticides are neurotoxic compounds widely used around the world. Evaluation of OP exposure in human studies is important for enabling adequate data analyses and drawing accurate conclusions. The aim of this study was to analyze OP exposure biomarkers and their relationships in a Mexican population with different exposure levels. Dialkylphosphates (DAP) were determined through gas chromatography-mass spectrometry (GC-MSD); acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), arylesterase (AREase), basal paraoxonase 1 (PONase), and ß-glucuronidase activities were detected using spectrophotometric methods. The albumin content was determined in a certified clinical laboratory. The DMTP metabolite was found in the highest concentration, and a negative and significant correlation between DAP and cholinesterase activity was observed. Our results suggested that BuChE is a considerably more sensitive biomarker than AChE. In addition, ß-glucuronidase was positively correlated with albumin, BuChE, and PONase. In conclusion, our data strongly support the use of two or more biomarkers of exposure in human monitoring and the application of a strong and validated questionnaire.

Relationship between butyrylcholinesterase activity and lipid parameters in workers occupationally exposed to pesticides.

Exposure to organophosphate pesticides (OP) has been associated with the inhibition of cholinesterase enzymatic activity, such as butyrylcholinesterase (BuChE). Changes in BuChE activity have been associated with obesity, diabetes, hyperthyroidism, and metabolic syndrome. However, few studies have evaluated the effects of pesticides on both BuChE and lipid parameters. The aim of this study was to evaluate lipid parameters in urban sprayers and their association with BuChE activity. An analytical cross-sectional study was conducted in workers exposed to pesticides. The pesticide exposures were evaluated by the measurement of urinary dialkylphosphates. BuChE activity was determined spectrophotometrically in serum, and biochemical parameters were determined at a certified laboratory. Information regarding general characteristics, lifestyle, and other aspects was obtained from a structured questionnaire. The results showed variations in glucose, cholesterol, albumin, atherogenic index, creatinine, LDL, VLDL, triglycerides, and total lipids according to the level of exposure to pesticides in individuals with overweight and obesity. Furthermore, positive correlations between BuChE activity and lipid parameters were observed; these effects were associated with the body mass index. More studies are needed in human population to better elucidate the role of BuChE in lipid metabolism.

In vitro inhibition of human red blood cell acetylcholinesterase (AChE) by temephos-oxidized products.

Temephos (Tem) is an organophosphorus pesticide widely used to kill and prevent the growth of the main vectors for the transmission of dengue, zika, and chikungunya viruses. In chlorinated water, Tem is oxidized to its dioxon-sulfoxide (Tem-dox-SO), dioxon-sulfone (Tem-dox-SO2), and sulfoxide (Tem-SO) derivatives; however, these compounds are not commercially available to be used as standards and in toxicological studies. In the present study, we synthesized and characterized the Tem-oxidation products and the compound 4,4'-sulfinyldiphenol. These compounds were obtained by a simple reaction between Tem or 4,4'-thiodiphenol with sodium hypochlorite or potassium periodate, and were characterized by IR, NMR, and UPLC-HRESIMS. The in vitro evaluation of inhibitory potency of Tem-oxidized products on human red blood cell acetylcholinesterase (RBC AChE) showed that Tem-dox-SO2 was the most potent inhibitor of human RBC AChE, and its effect was more pronounced than that observed for ethyl-paraoxon, a potent typical inhibitor of AChE. An HPLC-DAD method for the analysis of metabolic products of Tem was developed, which may be useful for monitoring in biological and environmental samples. The ability of Tem-oxidized metabolites to inhibit human RBC AChE suggests that the addition of Tem to chlorinated drinking water could result in an increase in the risk of RBC AChE inhibition after exposure.

Relationship between internal and external factors and the activity of PON1.

Paraoxonase 1 (PON1) is an A-esterase calcium-dependent enzyme that is associated with high-density lipoprotein (HDL) and capable of hydrolyzing a wide variety of substrates, including organophosphate (OP) pesticides. The PON1 phenotype can be modulated by multiple internal and external factors, thereby affecting the catalytic capacity of the enzyme. The aim of this study was to evaluate factors that could modulate PON1 activity in a sample occupationally exposed to pesticides. A cross-sectional, descriptive, and analytical study was carried out with 240 workers. The participants were stratified according to their level of pesticide exposure as reference, moderate-exposure, and high-exposure groups. PON1 activities (arylesterase/AREase, CMPAase, and ssPONase (salt-stimulated)) were determined by spectrophotometry, and the Q192R and L55MPON1 genotypes by real-time PCR. The most frequent genotypes were heterozygous (QR) and homozygous (LL) for PON1Q192R and PON1L55M polymorphisms, respectively. The internal factors associated with the activity of PON1 were the PON1 genotypes (55 and 192) and biochemical parameters related to the lipid profile, in contrast, various external factors related to diet and harmful habits as well as with exposure to pesticides were associated with the activity of PON1. However, using a multivariate mixed ordinal regression model, we found a significant reduction of ssPONase activity in the high-exposure group compared with the reference group only in haplotypes QQLL and RRLL.

Trends on deaths from acute pesticide poisoning in Mexico, 2000-2021 / Tendências das mortes por intoxicação aguda por pesticidas no México, 2000-2021

ABSTRACT Objetive: To provide a comprehensive analysis of mortality trends from acute pesticide poisoning in Mexico from 2000 through 2021. Methods: The governmental records of deaths from acute pesticide poisoning were used. The age-standardized years of life lost and aged-standardized mortality rates were estimated. Significant changes in trends of annual percentage change were identified using Joinpoint regression. Results: Between 2000 and 2021, mortality was primarily observed in individuals aged 15 to 19 years. Males were the most affected. Self-inflicted pesticide poisoning was the primary registered reason for death. The age-standardized mortality rate from acute pesticide poisoning was reduced from 2012 to 2021 (APC: -4.4; p=0.003). Conclusion: This report is the first study about the mortality rate from acute pesticide poisoning in Mexico. The results provided evidence to consider in developing laws to prevent acute pesticide poisoning.

RESUMO Objetivo: Fornecer uma análise abrangente das tendências de mortalidade por envenenamento agudo por pesticidas no México de 2000 a 2021. Métodos: Foram usados os registros governamentais de mortes por envenenamento agudo por pesticidas. Foram estimados os anos de vida perdidos estandardizados por idade e as taxas de mortalidade estandardizados por idade. Modificações significativas nas tendências de variação percentual anual foram identificadas usando a regressão Joinpoint. Resultados: Entre 2000 e 2021, a mortalidade foi observada principalmente em indivíduos na faixa etária de 15 a 19 anos. Os homens foram os mais afetados. O envenenamento por pesticida autoinfligido foi o principal motivo de morte registrado. A taxa de mortalidade estandardizada por idade por intoxicação aguda por pesticidas foi reduzida de 2012 a 2021 (Annual Percent Change — APC: -4,4; p=0,003). Conclusão: Este relatório é o primeiro estudo sobre a taxa de mortalidade por intoxicação aguda por pesticidas no México. Os resultados forneceram evidências a serem consideradas no desenvolvimento de leis para prevenir o envenenamento agudo por pesticidas.

Modified CDKN2B (p15) and CDKN2A (p16) DNA methylation profiles in urban pesticide applicators.

Gene-specific changes in DNA methylation by pesticides in occupationally exposed populations have not been studied extensively. Of particular concern are changes in the methylation profile of tumor-suppressor, such as CDKN2B and CDKN2A, genes involved in oncogenesis. The aim of this study was to evaluate the methylation profiles of CDKN2B and CDKN2A genes in urban pesticide applicators and their relationship with occupational exposure to pesticides. A cross-sectional study was conducted in 186 urban pesticide applicators (categorized as high or moderate exposures) and 102 participants without documented occupational exposures to pesticides. Acute and chronic pesticide exposures were evaluated by direct measurement of urinary dialkylphosphates, organophosphate metabolites, and a structured questionnaire, respectively. Anthropometric characteristics, diet, clinical histories, and other variables were estimated through a validated self-reported survey. DNA methylation was determined by pyrosequencing of bisulfite-treated DNA. Decreased DNA methylation of the CDKN2B gene was observed in pesticide-exposed groups compared to the non-exposed group. In addition, increased methylation of the CDKN2A promoter was observed in the moderate-exposure group compared to the non-exposed group. Bivariate analysis showed an association between CDKN2B methylation and pesticide exposure, general characteristics, smoking status, and micronutrients, while changes in CDKN2A methylation were associated with pesticide exposure, sex, educational level, body mass index, smoking status, supplement intake, clinical parameters, and caffeine consumption. These data suggest that pesticide exposure modifies the methylation pattern of CDKN2B and CDKN2A genes and raise important questions about the role that these changes may play in the regulation of cell cycle activities, senescence, and aging.