Available via license: CC BY
Content may be subject to copyright.
Article Not peer-reviewed version
Pioneering Insights: The Intersection of
Pesticide Exposure, Genetic Variations,
and Health Risks in Southeastern
Brazilian Farmers
Débora Dummer Meira , Victor Nogueira da Gama Kohls , Matheus Correia Casotti , Luana Santos Louro ,
Gabriel Mendonça Santana , Thomas Erik Santos Louro , Adriana Madeira Álvares da Silva ,
Lorena Souza Castro Altoé , Raquel Silva dos Reis Trabach , Sonia Groisman , Elizeu Fagundes de Carvalho ,
Jamila Alessandra Perini Machado , Stephanie Seneff * , Iúri Drumond Louro
Posted Date: 31 July 2024
doi: 10.20944/preprints202407.2592.v1
Keywords: CYP2C9; Pesticides; Glyphosate; Cardiovascular Disease; Attention Deficit; Hypertension
Preprints.org is a free multidiscipline platform providing preprint service that
is dedicated to making early versions of research outputs permanently
available and citable. Preprints posted at Preprints.org appear in Web of
Science, Crossref, Google Scholar, Scilit, Europe PMC.
Copyright: This is an open access article distributed under the Creative Commons
Attribution License which permits unrestricted use, distribution, and reproduction in any
medium, provided the original work is properly cited.
Article
Pioneering Insights: The Intersection of Pesticide
Exposure, Genetic Variations, and Health Risks in
Southeastern Brazilian Farmers
Débora Dummer Meira 1,†, Victor Nogueira Da Gama Kohls 1, †, Matheus Correia Casotti 1,
Luana Santos Louro 2, Gabriel Mendonça Santana 2, Thomas Erik Santos Louro 3,
Adriana Madeira Alvares da Silva 4, Lorena Souza Castro Altoé 1, Raquel Reis Trabach 1,
Sonia Groisman 5, Elizeu Fagundes de Carvalho5, Jamila Alessandra Perini Machado 6,
Stephanie Seneff 7,* and Iúri Drumond Louro 1
1 Núcleo de Genética Humana e Molecular, Universidade Federal do Espírito Santo (UFES), Espírito Santo,
Brasil.
2 Centro de Ciências da Saúde, Curso de Medicina, Universidade Federal do Espírito Santo (UFES), Espírito
Santo, Brasil.
3 Escola Superior de Ciências da Santa Casa de Misericórdia de Vitória (EMESCAM), Espírito Santo, Brasil.
4 Departamento de Morfologia, Universidade Federal do Espírito Santo (UFES), Espírito Santo, Brasil.
5 Instituto de Biologia Roberto Alcântara Gomes (IBRAG), Universidade do Estado do Rio de Janeiro (UERJ),
Rio de Janeiro, Brasil.
6 Laboratório de Pesquisa de Ciências Farmacêuticas, Universidade do Estado do Rio de Janeiro (UERJ), Rio
de Janeiro, Brasil.
7 Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology,
Cambridge MA USA 02139.
* Correspondence: seneff@csail.mit.edu; Computer Science and Artificial Intelligence Laboratory, MIT,
Cambridge MA USA 02139.
† Both authors contributed equally to the writing of this manuscript.
Abstract: Brazil is the world leader in pesticide consumption, and its indiscriminate use puts farmers’ health
at risk. The CYP2C9 gene stands out for encoding the CYP2C9 enzyme, which metabolizes several endogenous
substrates and specific xenobiotics, especially pesticides. Our goal is to study the risk of pesticide use, especially
the herbicide glyphosate, in the development of diseases and the association with two CYP2C9 polymorphisms,
in farmers living in the southern region of Espírito Santo state, Brazil. The allelic frequency of CYP2C9*1,
CYP2C9*2 and CYP2C9*3 was determined in blood samples from individuals exposed or not to pesticides using
real-time PCR. 304 blood samples were analyzed, dividing CYP2C9 genotypes into three metabolization
classes: normal, intermediate, and slow. Our results show that normal metabolizers are more vulnerable to
diseases such as blood pressure changes and cardiovascular and kidney problems, while intermediate
metabolizers show an association with attention deficit disorder and miscarriages, suggesting that farmers in
the studied region developed symptoms correlated to the CYP2C9 genotype they have. This work is pioneering
in associating specific genetic variations and health risks with pesticide exposure, emphasizing the importance
of personalized medicine and stricter regulation of pesticide use for public health and occupational safety.
Keywords: CYP2C9; pesticides; glyphosate; cardiovascular disease; attention deficit; hypertension
1. Introduction
Brazil leads the world rank in pesticide consumption, a worrying fact that is directly related to
the country's agricultural policy, focused on increasing productivity to compete in the international
market [1]. Excessive use of pesticides, often without due technical information and without the
adequate use of personal protective equipment (PPE), exposes farmers, their families and even the
end consumer to significant health risks [2–5]. Substances widely used in agriculture, such as
Disclaimer/Publisher’s Note: The statements, opinions, and data contained in all publications are solely those of the individual author(s) and
contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting
from any ideas, methods, instructions, or products referred to in the content.
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
© 2024 by the author(s). Distributed under a Creative Commons CC BY license.
2
organophosphates, carbamates, malthion, diazinone, and especially glyphosate, have been classified
as probable carcinogens by the International Agency for Research on Cancer [6–8]. The Cytochrome
P450 family of enzymes, including CYP2C9, is essential for xenobiotic metabolism, having a
characteristic molecular structure that allows the metabolization of several substances [9–11].
Recent studies have changed the perception about pesticide toxicity to humans, with evidence
that substances such as glyphosate can inhibit the activity of the CYP2C9 enzyme [12–14].
Furthermore, there is an association between central nervous system neoplasms and chemically
exposed occupations, such as agriculture [15–17]. Symptoms of pesticide poisoning can range from
acute effects, such as headaches and nausea, to chronic conditions, including liver and kidney
damage [18,19].
In this study, the impact of pesticides on the development of diseases/symptoms in farmers of
the south of Espírito Santo was evaluated, focusing on the relationship between daily exposure and
CYP2C9 polymorphisms and seeking to correlate the emergence or worsening of diseases with these
polymorphisms, analyzing a total of 43 variables related to diseases and symptoms.
2. Materials and Methods
2.1. Study population
This research was submitted to the research ethics committee of the Federal University of
Espírito Santo and approved according to opinion number 3.378.510 and the resolutions of the
National Health Council nº 466 (of December 12, 2012) and nº 510 (of April 7, 2016). Inclusion criteria
were men and women over 18 years of age, resident farmers of the south of the Espírito Santo state.
A questionnaire was applied to each volunteer, and then blood was collected. DNA extraction and
genotyping of 304 DNA samples were analyzed, subdivided into two groups: YES - Farmers who use
pesticides (case/test) and NO - Farmers who do not use pesticides (control).
2.2. DNA extraction
DNA extraction and purification were carried out using a Qiagen® commercial kit, following
manufacturer’s recommendations (Qiagen, USA).
2.3. Real-Time PCR to Assess CYP2C9 Polymorphisms
CYP2C9 polymorphisms were identified by real-time PCR as previously described by Perini et
al., (2009). Validated TaqMan® assays (Applied Biosystems, USA) were used to discriminate
CYP2C9*2 (rs1799853) and CYP2C9*3 (rs1057910) alleles. For both assays, real-time PCR reactions
were performed in a final volume of 10 μl containing 30 ng of DNA, 1X Taqman Universal Master
Mix (Applied Biosystems, USA), 1X of each specific assay and H2O q.s.p. PCR conditions were initial
denaturation at 95 °C for 10 minutes, followed by 40 cycles of denaturation at 92 °C for 15 seconds
and annealing/extending at 60 °C for 1 minute. All samples were analyzed using the Fast 7500 Real-
Time System (Applied Biosystems, USA). Genotypes were determined by analyzing the allelic
discrimination plots. According to each sample’s alleles, individuals were classified into different
categories regarding their metabolization capacity:
Category Metabolization Genotype(s)
A Normal *1/*1
B Intermediate *1/*2; *1/*3
C Slow *2/*2; *2/*3; *3/*3
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
3
It is important to emphasize the scarcity of cases in category C, making future analysis essential
for possible connections with experiments not addressed in this study. In this context, we focused
primarily on groups A and B, recognizing the need for comprehensive understanding to explore
potential correlations and broader implications.
2.4. Data analysis methodology
For bivariate statistical analysis, the Chi-square test and, when necessary, Fisher's exact test was
applied, with a 5% margin of error. To carry out the analyses, the free R version 3.6.1 software was
used. The software can be obtained from the website https://cran.r-project.org/bin/windows/base/.
Furthermore, all results derive from the bivariate analysis between “use of pesticides” and
“appearance of a certain symptom” for each genotype group.
3. Results
Allelic and genotypic frequencies were calculated and presented in Tables 1 and 2.
Table 1. Genotypic frequencies (N=304).
Genotypes n Frequency
*1/*1 196 0.645
*1/*2 68 0.224
*1/*3 28 0.092
*2/*2 5 0.016
*2/*3 5 0.016
*3/*3 2 0.007
N: total number of individuals; n: sample subtotal; Frequency=n/N.
Table 2. - Allelic frequencies (N=608).
Allele n Frequency
*1 488 0.803
*2 83 0.137
*3 37 0.061
N: total number of individuals; n: sample subtotal; Frequency=n/N.
Most of interviewed workers (72.01%) were males and 27.99% were females. Farmers were
predominantly aged between 34 and 50 years, having worked as such for 26 to 50 years, and 86 of
them being in both groups (32.21%).
The percentage of rural workers who use pesticides was as high as 90.27%. Glyphosate was the
most used (79.66%), followed by flutriafol (32.85%), thiamethoxam (26.91%) and cyproconazole
(22.34%). Behavioral variables such as alcohol consumption and tobacco use were not correlated with
genotypes and diseases/symptoms emergence.
Symptoms/diseases addressed were eye irritation, skin lesions/allergies, skin burns,
nausea/vomiting, phlegm, abdominal pain, diarrhea, difficult digestion, wheezing, asthma, gastric
inflammations, liver diseases, blood pressure changes, infertility, tearing, dizziness/vertigo, cough,
shortness of breath/dyspnea, blurred vision, tremors, vomiting, joint pain, hepatitis,
osteoarthritis/osteoporosis, kidney diseases, respiratory diseases, attention deficit, miscarriages,
headache, excessive sweating, salivation, agitation/irritability, tingling, miosis, cramps, body/muscle
pain, depression, cardiovascular diseases, cancer, heart palpitations, salivary gland damage,
malformation and hyperactivity. Significant correlations (P<0.05) between the use of pesticides and
symptoms/diseases were tested for each genotype group (“A”, “B” and “C”).
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
4
As shown in Table 3, “B” intermediate metabolizers (who use pesticides) are associated with
attention deficit (Chi-square P = 0.01782 / Fisher P = 0.02008) and miscarriages (Chi-square P = 0.00070
/ Fisher P = 0.00409), and in normal “A” metabolizers (who use pesticides) an association with blood
pressure alteration (Chi-square P = 0.00371 / Fisher P = 0.00305), cardiovascular diseases (Chi-square
P = 0.04308 / Fisher P = 0.03319) and kidney diseases (Chi-square P = 0.06536 / Fisher P = 0.02635) was
found.
Table 3. Diseases and symptoms of intermediate metabolizers “B” and normal metabolizers “A” of
CYP2C9.
Disease/Symptoms Group P-value
(Chi-square)
P-value
(Fisher)
Blood pressure alterations A 0.00371
0.00305
Cardiovascular diseases A 0.04308
0.03319
Kidney diseases A 0.06536
0.02635
Attention deficit B 0.01782
0.02008
Miscarriages B 0.00070
0.00409
4. Discussion
In this paper, we have investigated a possible link between exposure to glyphosate and other
pesticides among agricultural workers in Brazil and various health issues and have explored
specifically whether different polymorphisms of the CYP2C9 liver enzyme might play a role in how
pesticides impact health. We have found a statistically significant increase in blood pressure issues
(hypotension/hypertension), cardiovascular disease, kidney disease, attention deficit, and
miscarriages, when comparing pesticide-exposed agricultural workers with controls who worked on
organic farms, with interesting differences observed between normal metabolizers (*1 alleles only)
and intermediate metabolizers (*1 and either *2 or *3). In particular, normal metabolizers (group A)
are more susceptible to hypertension, cardiovascular disease and kidney disease, whereas
intermediate metabolizers (group B) are more susceptible to attention deficit and miscarriages. Sparse
data prevented us from reaching any conclusions about group C (slow metabolizers), who had *2
and/or *3 alleles.
Glyphosate stands out as the most used pesticide by Espírito Santo farmers. According to
Boocock et al. [20], glyphosate inhibits plant growth by interfering with the production of essential
aromatic amino acids, primarily through suppression of the enzyme 5-enolpyruvylshikimate-3-
phosphate synthase (EPSPS). In the terrestrial environment, glyphosate is mainly biodegraded into
aminomethylphosphonic acid (AMPA). When metabolized by soil bacteria, glyphosate can cause
several toxicological problems if absorbed. It can leak into groundwater and be transformed into
formaldehyde, which is carcinogenic and neurotoxic [21]. Another problem with glyphosate-based
herbicides is the presence of N-nitrosoglyphosate (NNG) as a contaminant (0.1 ppm), which is a
highly carcinogenic substance [21]. NNG can also be formed in soil, water or in the human body
when glyphosate combines with nitrates and nitrites [21].
Glyphosate is a glycine molecule with a methyl-phosphonyl group attached to the nitrogen
atom. By acting as a glycine analogue, it is possible that glyphosate can displace glycine at random
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
5
points during protein synthesis, with unknown consequences. Several synthetically produced amino
acids, close structural analogues of natural amino acids, can be erroneously incorporated into
peptides [22,23]. The correlations between glyphosate use and the recent alarming rise in several
modern diseases are striking, as presented by Swanson et al. [24] These include obesity, diabetes,
end-stage renal disease, kidney failure, autism, Alzheimer’s disease, dementia, Parkinson’s disease,
multiple sclerosis, intestinal infection, inflammatory bowel disease, stroke, leukemia, thyroid cancer,
liver cancer, pancreatic cancer and kidney cancer [24] .
We found a highly statistically significant correlation with pesticide use among Group A farmers
for blood pressure alterations (p = 0.003). As early as 1974, hypertension had been identified as the
most common and most potent contributor to cardiovascular mortality [25]. There is a 30-55%
prevalence of hypertension in the general population in Europe, with higher rates in the elderly [26].
As of 2018, half the people in the United States over 20 years old suffers from hypertension [27]. A
study based in China evaluated health risks of glyphosate exposure in factory workers involved in
the production of glyphosate. Compared to workers at the same company who were not exposed to
glyphosate, the exposed group had statistically significant higher rates of hypertension, coronary
artery disease, elevated liver enzymes, and renal disease [28].
de Marins et al. [29], Ojelade et al. [30] and Gress et al. [31] have published literature reviews
addressing potential cardiac issues associated with glyphosate exposure, and they found evidence of
long QT syndrome, conduction blocks, arrhythmias, and cardiac arrest in cases of acute glyphosate
exposure. These studies delved into the potential arrhythmogenic mechanism of glyphosate in
mammalian cardiac tissues. Considering the rising use of synthetic molecules in agriculture, the
review by Ojelade et al.[30] highlighted the adverse impact on human health and ecosystems [30].
Recently, the enzyme 21-hydroxylase, encoded by the CYP21A2 gene, was implicated in steroid
hormone synthesis, suggesting that glyphosate’s inhibition of CYP2C9 could impact endocrine
regulation and influence farmers’ blood pressure [32]. Glyphosate-based herbicides, widely used in
woodlands and farmlands, have raised concerns due to toxicological issues of glyphosate and its
metabolite AMPA in the food chain. Glyphosate has been linked to various health problems,
necessitating a comprehensive review of its use, associated risks, and maximum residue limits. The
compilation of such data aims to guide regulatory agencies in advising safe glyphosate usage
practices [30].
Catani et al. [33] have shown that perinatal exposure of rats to glyphosate causes oxidative
damage in the brain associated with reduced melatonin levels. Low urinary melatonin levels are
associated with essential hypertension [34]. Melatonin supplementation in the evening has been
shown to reduce nighttime blood pressure in men suffering from essential hypertension [35].
Gunatilake et al. [36] proposed that glyphosate, even without added formulators, has a unique
insidious mechanism of toxicity that involves the erroneous substitution for the coding amino acid
glycine during protein synthesis, which in certain circumstances can lead to nearly complete
inactivation of the affected protein’s enzymatic activity. Glyphosate’s inhibition of cytochrome P450
enzymes in the liver would disrupt the liver’s ability to detoxify and eliminate fat soluble toxic
exposures, including pathogenic metabolites, toxic environmental chemicals, and prescription drugs
[37], causing them to be much more nephrotoxic than they would normally be [38].
In this study, we observed that farmers classified as intermediate metabolizers (B) who used
pesticides presented more “attention deficit” when compared to farmers who did not use pesticides.
Although we did not find a direct correlation with this symptom in the literature, we observed that
several researchers have found correlations between the use of pesticides and psychiatric diseases
such as autism and Alzheimer’s disease [39,40].
Attention deficit hyperactivity disorder (ADHD) has been conceptualized as a childhood
disorder that diminishes with age. However, there is less awareness of adult ADHD as a condition,
but a systematic study determined that 2.58% of the population worldwide continues to have
symptoms of ADHD even after they age out of a childhood diagnosis, and, overall, 6.76% of the global
adult population suffers from symptomatic adult ADHD [41]. A study of the trends over time in the
United States of ADHD prevalence (according to US Centers for Disease Control and Prevention
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
6
(CDC) data) showed a rising trend over time from 1990 to 2010, which correlated very strongly with
the rising use of glyphosate on corn and soy crops (R = 0.9466, p ≤ 0.000036) [42]. While correlation
does not necessarily mean causation, there is an increasing number of peer-reviewed publications
showing evidence that glyphosate causes specific pathologies that are linked with ADHD.
The incidence of ADHD in children is rising rapidly, from about 12 cases per 1000 people 30
years ago to about 35 cases per 1000 people in the late 1990s [43]. Learning and memory impairment
are common ADHD symptoms. While ADHD has become very common in both children and adults,
the causes of ADHD and the underlying brain pathology are still poorly understood. Recently,
evidence is building to support the idea that ADHD is caused by an interaction between genetics and
environmental factors linked to the Wnt and mammalian Target of Rapamycin (mTOR) signaling
pathways, both of which are heavily involved in neurodevelopment [44]. Glyphosate exposure
during pregnancy in rat dams caused suppression of the Wnt signaling pathway in exposed embryos,
leading to behavioral and cognitive issues in the offspring [45]. Glyphosate exposure during
pregnancy and lactation leads to abnormalities in the Wnt/β-catenin and Notch pathways in the
prefrontal cortex of mouse offspring, potentially contributing to neurodevelopmental disorders [46].
Glyphosate has been linked to gut dysbiosis through suppression of critical beneficial microbes
and overgrowth of pathogenic species. Pathogenic Clostridia species can produce toxic metabolites
that cause neurological deviations in the brain, particularly when CYP enzymes are compromised
[47]. Imbalances in the gut microbiome observed in association with ADHD could be related to
impairments in dopaminergic signaling [48].
Glyphosate exposure in rats causes neurotoxicity by altering serotonergic, dopaminergic, and
noradrenergic systems, with dose-related changes in neurotransmitter levels [49]. As early as 1999,
dysregulation of central noradrenergic networks has been linked to ADHD [50]. These networks are
involved in modulating high level cortical functions such as attention, alertness, vigilance, and
executive function. Furthermore, disruptions in the dopaminergic system, particularly genetic
defects in critical genes involved in dopamine signaling regulation, have been implicated in ADHD
[51]. Genetic mutations involved in serotonin signaling have also been implicated in the etiology of
ADHD [52]. In a machine learning model, serotonin transporter gene SNPs were linked to ADHD
[53].
Arnsten et al. hypothesized in 2009 that a weakness in the prefrontal association cortex
characterizes ADHD. This center is particularly dependent on dopaminergic stimulation and
adrenergic stimulation for proper function. Stimulants that have been widely used to treat ADHD
specifically enhance catecholamine signaling in the prefrontal cortex, increasing expression of both
dopamine and norepinephrine [54].
Glyphosate has been shown experimentally to cause melatonin deficiency. Glyphosate exposure
to rats prenatally and perinatally caused a 43% reduction in melatonin serum levels measured after
pups had matured, likely through epigenetic effects [33]. The mechanism might be traced to the
shikimate pathway, which glyphosate suppresses in gut microbiota. Melatonin is derived from the
amino acid tryptophan, which is one of the three aromatic amino acids synthesized by gut microbes
via the shikimate pathway. In vitro studies with rat pinealocytes exposed to 50 μm glyphosate showed
that glyphosate activated metabotropic glutamate receptors, and that this caused a reduction in
melatonin synthesis [33]. Metabotropic glutamate receptor activation suppresses melatonin synthesis
in rat pinealocytes [55].
According to a review paper published in 2022, glyphosate exposure can cause many neurotoxic
effects, affecting cell development, neurotransmission, and causing neuronal death, and behavioral
and motor disorders in humans, rodents, fish, and invertebrates [56]. All these disruptions could
adversely impact brain development, leading to symptoms of ADHD. Based on an extensive
literature review, Seneff et al. [40] provided a theoretical argument for how glyphosate's suppression
of melatonin synthesis and induction of oxidative stress in the brain through glutamate
neuroexcitotoxicity could lead to the neurodevelopmental defects associated with autism.
Glyphosate exposure to neurons disrupts synaptic assembly, and glyphosate reduces synaptic
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
7
protein expression in the hippocampus, likely contributing to cognitive impairment observed in
glyphosate-exposed developing rats [57].
Through genetic studies, deficiencies in melatonin signaling have been found in association with
ADHD. In one experiment, 101 patients with ADHD were compared to 220 controls from the general
population. Several damaging mutations were found in patients with ADHD. A specific mutation in
N-acetylserotonin O-methyltransferase (ASMT) and another in melatonin receptor 1A (MTNR1A)
were detected exclusively in ADHD patients, and both of these mutations were determined to abolish
enzyme activity [58]. Melatonin therapy has been found to be beneficial for ADHD patients,
particularly for treating insomnia [59].
One of the mechanisms involved in the emergence of these diseases is the ability of glyphosate
to chelate metals. A study on Danish dairy cattle investigated the mineral composition in the serum
of cattle fed feed containing traces of glyphosate in which cobalt (Co) and manganese (Mn) were
deficient [14,60,61]. According to Samsel and Seneff [23], manganese (Mn) is one of the 14 essential
trace elements in the human body, and its deficiency can explain pathologies associated with
glyphosate use, notably autism and Alzheimer’s disease, as well as attention deficit [62,63]. Another
concern is that the glyphosate breakdown product AMPA can leak into groundwater and be
transformed into formaldehyde, which is neurotoxic [21].
In this study, correlations were found between the use of pesticides and spontaneous abortions
in group “B” of CYP2C9 genotypes (intermediate metabolizers). Swanson et al.[24] showed that the
endocrine disrupting properties of glyphosate can lead to reproductive problems, such as infertility,
miscarriage, birth defects and changes in sexual development. Glyphosate has also been linked to
polycystic ovary syndrome, a major factor in female infertility, through its estrogenic properties [64].
Muñoz published a review paper showing that glyphosate has the key characteristics of an endocrine
disruptor, based on its observed toxic effects [65].
Vianna-Jorge et al. [66] and Perini et al. [67] describe the frequencies of *1, *2 and *3 alleles in the
Brazilian population, which was in accord with the frequencies we found in this study. Our results
show that normal metabolizers “A” for CYP2C9 (*1/*1) are more susceptible to developing a
disease/symptom in relation to individuals carrying polymorphic variants of metabolizer groups “B”
(*1/*2, *1/*3). To date, no metabolizing enzymes (including all CYPs) have been described to
metabolize glyphosate in mammals. Secondary glyphosate metabolites generated by soil and
intestinal bacteria, as well as glyphosate itself, have high toxicity [68]. The next step of this research
should include the evaluation of CYP2C9 polymorphism in the general population to verify a
correlation with symptoms/diseases observed in Espírito Santo farmers. Additionally, we would like
to evaluate other diseases, particularly neurodevelopmental diseases such as autism, for their
possible association with pesticides [47,62].
5. Conclusion
We have found that individuals who use pesticides and have CYP2C9 “A” normal metabolizer
(*1/*1) are more likely to develop diseases and symptoms compared to those who carry polymorphic
variants “B" (*1/*2, *1/*3). We observed that Espírito Santo farmers exposed to pesticides manifested
changes in blood pressure, cardiovascular and kidney diseases, miscarriages and attention deficit,
correlating with their CYP2C9 genotypes. Continued research about the impacts of pesticide use on
global health and the role of genetics in altering susceptibility will help guide new pesticide use
policies.
6. Patents
Not applicable.
Supplementary Materials: Not applicable.
Author Contributions: Conceptualization, Débora Meira and Victor Kohls; Data curation, Débora Meira and
Victor Kohls; Methodology, Débora Meira , Victor Kohls, Adriana da Silva and Jamila Alessandra Machado;
Supervision, Iúri Louro; Validation, Adriana da Silva , Jamila Alessandra Machado and Iúri Louro;
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
8
Visualization, Raquel Trabach , Sonia Groisman, Elizeu de Carvalho, Stephanie Seneff and Iúri Louro; Writing –
original draft, Débora Meira , Victor Kohls and Stephanie Seneff; Writing – review & editing, Matheus Casotti ,
Luana Louro, Gabriel Santana, Thomas Erik Louro, Lorena Altoé, Raquel Trabach , Sonia Groisman, Elizeu de
Carvalho and Stephanie Seneff. All authors have read and agreed to the published version of the manuscript.
Funding: Financial support was provided by grants from the Brazilian agencies: Fundação Carlos Chagas Filho
de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico
e Tecnológico (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES). This
study was also funded by the Foundation for Research and Innovation Support of the State of Espírito Santo –
FAPES through the Research Programs for SUS – PPSUS 10/2013 (grant number: 74713515/2016) and PPSUS
05/2015 (grant number: 65883616/2014). Stephanie Seneff was funded in part by Quanta Computer, Inc. in
Taiwan (grant number: 6897576).
Institutional Review Board Statement Not applicable.
Informed Consent Statement: Not applicable.
Ethical approval: This work was approved by the Research Ethics Committee (CEP) of the Federal University
of Espírito Santo and under opinion number 3.378.510 and resolutions of the National Health Council nº 466 (of
December 12, 2012) and nº 510 (April 7, 2016).
Data Availability Statement: Our ethics committee asks that we not share sensitive participant data. We are
willing to address any inquiries regarding the study methodology or results to the best of our ability. For
further information, please contact Débora Dummer Meira (debora.dummer.meira@gmail.com).
Acknowledgments: We would like to thank CAPES, CNPq, FAPES, FAPERJ and UFES (Federal University of
Espírito Santo) for supporting this project. We are grateful to Dr João Batista Pavesi Simão (Federal Institute of
Espírito Santo, Alegre, ES, Brazil) and we thank the farmers from the 22 rural communities who agreed to
participate in the project.
Conflicts of Interest: “The authors declare no conflicts of interest.”
Additional research: NCBI. Results for variants in the CYP2C9 gene.
<https://www.ncbi.nlm.nih.gov/clinvar/?term=CYP2C9%5Bgene%5D>.
Abbreviations
ADHD: Attention deficit hyperactivity disorder; AMPA: aminomethylphosphonic acid; CDC: Centers for
Disease Control and Prevention; Co: cobalt; EPSPS: 5-enolpyruvylshikimate-3-phosphate synthase; Mn:
manganese; mTOR: mammalian Target of Rapamycin; ASMT: N-acetylserotonin O-methyltransferase;
MTNR1A: N-acetylserotonin O-methyltransferase receptor 1A; NNG: N-nitrosoglyphosate; PPE: personal
Protection Equipment
References
1. Bombardi, L.M. Brazil: more cultivation, more pesticides, more exports Available online:
https://eu.boell.org/en/PesticideAtlas-Brazil (accessed on Sep 11, 2023).
2. Jobim, P.F.C.; Nunes, L.N.; Giugliani, R.; Cruz, I.B.M. da Existe uma associação entre mortalidade por
câncer e uso de agrotóxicos?: Uma contribuição ao debate. Cien. Saude Colet. 2010, 15, 277–288,
doi:10.1590/S1413-81232010000100033.
3. Pignati, W.A.; Lima, F.A.N. de S. e; Lara, S.S. de; Correa, M.L.M.; Barbosa, J.R.; Leão, L.H. da C.;
Pignatti, M.G. Distribuição espacial do uso de agrotóxicos no Brasil: uma ferramenta para a Vigilância
em Saúde. Cien. Saude Colet. 2017, 22, 3281–3293, doi:10.1590/1413-812320172210.17742017.
4. Latt, K.K.; Liana, S.R.; Poddar, S.; Bhaumik, A. Assessment of empty pesticide container disposal and
its effect on environment and human health: A review.; AIP Publishing, Ed.; AIP Conf. Proc, 2023; p.
060002, 10.1063/5.0163283
5. Lencucha, R.; Pal, N.E.; Appau, A.; Thow, A.-M.; Drope, J. Government policy and agricultural
production: a scoping review to inform research and policy on healthy agricultural commodities. Global.
Health 2020, 16, 11, doi:10.1186/s12992-020-0542-2.
6. Guyton, K.Z.; Loomis, D.; Grosse, Y.; El Ghissassi, F.; Benbrahim-Tallaa, L.; Guha, N.; Scoccianti, C.;
Mattock, H.; Straif, K. Carcinogenicity of tetrachlorvinphos, parathion, malathion, diazinon, and
glyphosate. Lancet Oncol. 2015, 16, 490–491, doi:10.1016/S1470-2045(15)70134-8.
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
9
7. Ore, O.T.; Adeola, A.O.; Bayode, A.A.; Adedipe, D.T.; Nomngongo, P.N. Organophosphate pesticide
residues in environmental and biological matrices: Occurrence, distribution and potential remedial
approaches. Environ. Chem. Ecotoxicol. 2023, 5, 9–23, doi:10.1016/j.enceco.2022.10.004.
8. Iwuozor, K.O.; Emenike, E.C.; Gbadamosi, F.A.; Ighalo, J.O.; Umenweke, G.C.; Iwuchukwu, F.U.;
Nwakire, C.O.; Igwegbe, C.A. Adsorption of organophosphate pesticides from aqueous solution: a
review of recent advances. Int. J. Environ. Sci. Technol. 2023, 20, 5845–5894, doi:10.1007/s13762-022-
04410-6.
9. Rang, H.P.; Dale, M.M.; Ritter, J.M.; Moore, P.K. Farmacologia; 5th ed.; Elsevier: São Paulo, 2003;
10. Singh, R.D.; Avadhesh, A.; Sharma, G.; Dholariya, S.; Shah, R.B.; Goyal, B.; Gupta, S.C. Potential of
Cytochrome P450, a Family of Xenobiotic Metabolizing Enzymes, in Cancer Therapy. Antioxid. Redox
Signal. 2023, 38, 853–876, doi:10.1089/ars.2022.0116.
11. Syed, K.; Mashele, S.S. Comparative Analysis of P450 Signature Motifs EXXR and CXG in the Large
and Diverse Kingdom of Fungi: Identification of Evolutionarily Conserved Amino Acid Patterns
Characteristic of P450 Family. PLoS One 2014, 9, e95616, doi:10.1371/journal.pone.0095616.
12. Abass, K.; Turpeinen, M.; Pelkonen, O. An evaluation of the cytochrome P450 inhibition potential of
selected pesticides in human hepatic microsomes. J. Environ. Sci. Heal. Part B 2009, 44, 553–563,
doi:10.1080/03601230902997766.
13. Elmadani, M.; Moustafa, Y.; El-Azab, M.; Pelkonen, O. Investigation of mechanism based inhibition of
CYP450 enzymes by selected organophosphorous, pyrethroids and benzoyl urea pesticides in pooled
human liver microsomes. Ther. Pharmacol. Clin. Toxicol. 2011, 15, 197–204.
14. Foldager, L.; Winters, J.F.M.; Nørskov, N.P.; Sørensen, M.T. Impact of feed glyphosate residues on
broiler breeder egg production and egg hatchability. Sci. Rep. 2021, 11, 19290, doi:10.1038/s41598-021-
98962-1.
15. Alavanja, M.C.R.; Bonner, M.R. Occupational Pesticide Exposures and Cancer Risk: A Review. J. Toxicol.
Environ. Heal. Part B 2012, 15, 238–263, doi:10.1080/10937404.2012.632358.
16. Clapp, R.W. Mortality among US employees of a large computer manufacturing company: 1969–2001.
Environ. Heal. 2006, 5, 30, doi:10.1186/1476-069X-5-30.
17. Nichols, L.; Sorahan, T. Cancer incidence and cancer mortality in a cohort of UK semiconductor
workers, 1970–2002. Occup. Med. (Chic. Ill). 2005, 55, 625–630, doi:10.1093/occmed/kqi156.
18. Augusto, L.G.S.; Carneiro, F.F.; Pignati, W.; Rigotto, R.M.; Friedrich, K.; Faria, N.M.X.; Búrigo, A.C.;
Freitas, V.M.T.; Guiducci Filho, E. Dossiê: Um alerta sobre os impactos dos agrotóxicos na saúde: agrotóxicos,
saúde, ambiente e sustentabilidade; 2nd ed.; Associação Brasileira de Saúde Coletiva: Rio de Janeiro, 2012.
19. Londres, F. Agrotóxicos no Brasil: um guia para ação em defesa da vida. Rio de Janeiro: Assessoria e
Serviços a Projetos em Agricultura Alternativa 2011, 191.
20. Boocock, M.R.; Coggins, J.R. Kinetics of 5-enolpyruvylshikimate-3-phosphate synthase inhibition by
glyphosate. FEBS Lett. 1983, 154, 127–133, doi:10.1016/0014-5793(83)80888-6.
21. Bohrer, L.C. Diagnóstico de riscos no uso de herbicidas nas lavouras de soja no município de Cruz Alta,
Universidade Federal de Santa Maria: Santa Maria, RS, 2005.
22. Rubenstein, E. Biologic Effects of and Clinical Disorders Caused by Nonprotein Amino Acids. Medicine
(Baltimore). 2000, 79, 80–89, doi:10.1097/00005792-200003000-00002.
23. Samsel, A.; Seneff, S. Glyphosate, pathways to modern diseases IV: cancer and related pathologies. J.
Biol. Phys. Chem. 2015, 15, 121–159, doi:10.4024/11SA15R.jbpc.15.03.
24. Swanson, N.; Leu, A.; Abrahamson, J.; Wallet, B. Genetically engineered crops, glyphosate and the
deterioration of health in the United States of America. J. Org. Syst. 2014, 9, 6–37.
25. Kannel, W.B. Role of blood pressure in cardiovascular morbidity and mortality. Prog. Cardiovasc. Dis.
1974, 17, 5–24, doi:10.1016/0033-0620(74)90034-6.
26. Kjeldsen, S.E. Hypertension and cardiovascular risk: General aspects. Pharmacol. Res. 2018, 129, 95–99,
doi:10.1016/j.phrs.2017.11.003.
27. Chobufo, M.D.; Gayam, V.; Soluny, J.; Rahman, E.U.; Enoru, S.; Foryoung, J.B.; Agbor, V.N.; Dufresne,
A.; Nfor, T. Prevalence and control rates of hypertension in the USA: 2017–2018. Int. J. Cardiol. Hypertens.
2020, 6, 100044, doi:10.1016/j.ijchy.2020.100044.
28. Pan, L.; Xu, M.; Yang, D.; Wang, B.; Zhao, Q.; Ding, E.-M.; Zhu, B. The association between coronary
artery disease and glyphosate exposure found in pesticide factory workers. J. Public Heal. Emerg. 2017,
1, 4–4, doi:10.21037/jphe.2016.12.04.
29. de Marins, M.L.R.; Nunes, J.A.; Da Silva Moraes, V.G.; de Lima, R.S.; de Oliveira Cardoso, M.V.; Araújo
Ribeiro, L.A. de; de Queiroz, D.B.; Silva, F.S. Maternal exposure to glyphosate-based herbicide causes
changes in the vascular function of offspring adult rats. Reprod. Toxicol. 2023, 115, 94–101,
doi:10.1016/j.reprotox.2022.12.004.
30. Ojelade, B.S.; Durowoju, O.S.; Adesoye, P.O.; Gibb, S.W.; Ekosse, G.-I. Review of Glyphosate-Based
Herbicide and Aminomethylphosphonic Acid (AMPA): Environmental and Health Impacts. Appl. Sci.
2022, 12.
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
10
31. Gress, S.; Lemoine, S.; Séralini, G.-E.; Puddu, P.E. Glyphosate-Based Herbicides Potently Affect
Cardiovascular System in Mammals: Review of the Literature. Cardiovasc. Toxicol. 2015, 15, 117–126,
doi:10.1007/s12012-014-9282-y.
32. Neves Cruz, J.; da Costa, K.S.; de Carvalho, T.A.A.; de Alencar, N.A.N. Measuring the structural impact
of mutations on cytochrome P450 21A2, the major steroid 21-hydroxylase related to congenital adrenal
hyperplasia. J. Biomol. Struct. Dyn. 2020, 38, 1425–1434, doi:10.1080/07391102.2019.1607560.
33. Cattani, D.; Pierozan, P.; Zamoner, A.; Brittebo, E.; Karlsson, O. Long-Term Effects of Perinatal
Exposure to a Glyphosate-Based Herbicide on Melatonin Levels and Oxidative Brain Damage in Adult
Male Rats. Antioxidants 2023, 12, 1825, doi:10.3390/antiox12101825.
34. Forman, J.P.; Curhan, G.C.; Schernhammer, E.S. Urinary melatonin and risk of incident hypertension
among young women. J. Hypertens. 2010, 28, 446–451, doi:10.1097/HJH.0b013e3283340c16.
35. Scheer, F.A.J.L.; Van Montfrans, G.A.; van Someren, E.J.W.; Mairuhu, G.; Buijs, R.M. Daily Nighttime
Melatonin Reduces Blood Pressure in Male Patients With Essential Hypertension. Hypertension 2004,
43, 192–197, doi:10.1161/01.HYP.0000113293.15186.3b.
36. Gunatilake, S.; Seneff, S.; Orlando, L. Glyphosate’s Synergistic Toxicity in Combination with Other
Factors as a Cause of Chronic Kidney Disease of Unknown Origin. Int. J. Environ. Res. Public Health
2019, 16, 2734, doi:10.3390/ijerph16152734.
37. Guengerich, F.P. Roles of cytochrome P450 enzymes in pharmacology and toxicology: Past, present,
and future. In Pharmacology and Toxicology of Cytochrome P450 – 60th Anniversary; Adv. Pharmacol. 2022;
pp. 1–47.
38. Fathy, Z.; El-Rahman, R.M.A. Effect of entomopathogenic nematodes Steinernema species
(steinernematidae: rhabditida) and Heterorhabditis bacteriophora (heterorhabditidae: rhabditida) on
the digestive enzymes and midgut histology of the African migratory locust Locusta migratoria migr.
Int. J. Trop. Insect Sci. 2023, 43, 727–736, doi:10.1007/s42690-023-00979-8.
39. Xu, Y.; Yang, X.; Chen, D.; Xu, Y.; Lan, L.; Zhao, S.; Liu, Q.; Snijders, A.M.; Xia, Y. Maternal exposure to
pesticides and autism or attention-deficit/hyperactivity disorders in offspring: A meta-analysis.
Chemosphere 2023, 313, 137459, doi:10.1016/j.chemosphere.2022.137459.
40. Seneff, S.; Kyriakopoulos, A.M.; Nigh, G. Is autism a PIN1 deficiency syndrome? A proposed etiological
role for glyphosate. J. Neurochem. 2024, Epub ahead of print. doi:10.1111/jnc.16140.
41. Song, P.; Zha, M.; Yang, Q.; Zhang, Y.; Li, X.; Rudan, I. The prevalence of adult attention-deficit
hyperactivity disorder: A global systematic review and meta-analysis. J. Glob. Health 2021, 11, 04009,
doi:10.7189/jogh.11.04009.
42. Seneff, S.; Swanson, N.; Li, C. Aluminum and Glyphosate Can Synergistically Induce Pineal Gland
Pathology: Connection to Gut Dysbiosis and Neurological Disease. Agric. Sci. 2015, 06, 42–70,
doi:10.4236/as.2015.61005.
43. Zhang, S.; Wu, D.; Xu, Q.; You, L.; Zhu, J.; Wang, J.; Liu, Z.; Yang, L.; Tong, M.; Hong, Q.; et al. The
protective effect and potential mechanism of NRXN1 on learning and memory in ADHD rat models.
Exp. Neurol. 2021, 344, 113806, doi:10.1016/j.expneurol.2021.113806.
44. Yde Ohki, C.M.; Grossmann, L.; Alber, E.; Dwivedi, T.; Berger, G.; Werling, A.M.; Walitza, S.; Grünblatt,
E. The stress–Wnt-signaling axis: a hypothesis for attention-deficit hyperactivity disorder and therapy
approaches. Transl. Psychiatry 2020, 10, 315, doi:10.1038/s41398-020-00999-9.
45. Coullery, R.; Pacchioni, A.M.; Rosso, S.B. Exposure to glyphosate during pregnancy induces
neurobehavioral alterations and downregulation of Wnt5a-CaMKII pathway. Reprod. Toxicol. 2020, 96,
390–398, doi:10.1016/j.reprotox.2020.08.006.
46. Ji, H.; Xu, L.; Wang, Z.; Fan, X.; Wu, L. Differential microRNA expression in the prefrontal cortex of
mouse offspring induced by glyphosate exposure during pregnancy and lactation. Exp. Ther. Med. 2017,
doi:10.3892/etm.2017.5669.
47. Rueda-Ruzafa, L.; Cruz, F.; Roman, P.; Cardona, D. Gut microbiota and neurological effects of
glyphosate. Neurotoxicology 2019, 75, 1–8, doi:10.1016/j.neuro.2019.08.006.
48. Shirvani-Rad, S.; Ejtahed, H.-S.; Ettehad Marvasti, F.; Taghavi, M.; Sharifi, F.; Arzaghi, S.M.; Larijani, B.
The Role of Gut Microbiota-Brain Axis in Pathophysiology of ADHD: A Systematic Review. J. Atten.
Disord. 2022, 26, 1698–1710, doi:10.1177/10870547211073474.
49. Martínez, M.-A.; Ares, I.; Rodríguez, J.-L.; Martínez, M.; Martínez-Larrañaga, M.-R.; Anadón, A.
Neurotransmitter changes in rat brain regions following glyphosate exposure. Environ. Res. 2018, 161,
212–219, doi:10.1016/j.envres.2017.10.051.
50. Biederman, J.; Spencer, T. Attention-deficit/hyperactivity disorder (adhd) as a noradrenergic disorder.
Biol. Psychiatry 1999, 46, 1234–1242, doi:10.1016/S0006-3223(99)00192-4.
51. Li, D.; Sham, P.C.; Owen, M.J.; He, L. Meta-analysis shows significant association between dopamine
system genes and attention deficit hyperactivity disorder (ADHD). Hum. Mol. Genet. 2006, 15, 2276–
2284, doi:10.1093/hmg/ddl152.
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
11
52. Quist, J.F.; Barr, C.L.; Schachar, R.; Roberts, W.; Malone, M.; Tannock, R.; Basile, V.S.; Beitchman, J.;
Kennedy, J.L. The serotonin 5-HT1B receptor gene and attention deficit hyperactivity disorder. Mol.
Psychiatry 2003, 8, 98–102, doi:10.1038/sj.mp.4001244.
53. Kautzky, A.; Vanicek, T.; Philippe, C.; Kranz, G.S.; Wadsak, W.; Mitterhauser, M.; Hartmann, A.; Hahn,
A.; Hacker, M.; Rujescu, D.; et al. Machine learning classification of ADHD and HC by multimodal
serotonergic data. Transl. Psychiatry 2020, 10, 104, doi:10.1038/s41398-020-0781-2.
54. Arnsten, A.F.T. ADHD and the Prefrontal Cortex. J. Pediatr. 2009, 154, I-S43,
doi:10.1016/j.jpeds.2009.01.018.
55. Yamada, H.; Yatsushiro, S.; Ishio, S.; Hayashi, M.; Nishi, T.; Yamamoto, A.; Futai, M.; Yamaguchi, A.;
Moriyama, Y. Metabotropic Glutamate Receptors Negatively Regulate Melatonin Synthesis in Rat
Pinealocytes. J. Neurosci. 1998, 18, 2056–2062, doi:10.1523/JNEUROSCI.18-06-02056.1998.
56. Costas-Ferreira, C.; Durán, R.; Faro, L.R.F. Toxic Effects of Glyphosate on the Nervous System: A
Systematic Review. Int. J. Mol. Sci. 2022, 23, 4605, doi:10.3390/ijms23094605.
57. Luna, S.; Neila, L.P.; Vena, R.; Borgatello, C.; Rosso, S.B. Glyphosate exposure induces synaptic
impairment in hippocampal neurons and cognitive deficits in developing rats. Arch. Toxicol. 2021, 95,
2137–2150, doi:10.1007/s00204-021-03046-8.
58. Chaste, P.; Clement, N.; Botros, H.G.; Guillaume, J.-L.; Konyukh, M.; Pagan, C.; Scheid, I.; Nygren, G.;
Anckarsäter, H.; Rastam, M.; et al. Genetic variations of the melatonin pathway in patients with
attention-deficit and hyperactivity disorders. J. Pineal Res. 2011, 51, 394–399, doi:10.1111/j.1600-
079X.2011.00902.x.
59. Heijden, K.B. Van der; Smits, M.G.; Someren, E.J.W. Van; Ridderinkhof, K.R.; Gunning, W.B. Effect of
Melatonin on Sleep, Behavior, and Cognition in ADHD and Chronic Sleep-Onset Insomnia. J. Am. Acad.
Child Adolesc. Psychiatry 2007, 46, 233–241, doi:10.1097/01.chi.0000246055.76167.0d.
60. Krüger, M.; Shehata, A.A.; Schrödl, W.; Rodloff, A. Glyphosate suppresses the antagonistic effect of
Enterococcus spp. on Clostridium botulinum. Anaerobe 2013, 20, 74–78,
doi:10.1016/j.anaerobe.2013.01.005.
61. Heymann, A.-K.; Schnabel, K.; Billenkamp, F.; Bühler, S.; Frahm, J.; Kersten, S.; Meyer, U.; von Soosten,
D.; Dänicke, S. Influences of Glyphosate Contaminations and Concentrate Feed on Performance, Blood
Parameters, Blood Cell Functionality and DNA Damage Properties in Fattening Bulls. Animals 2023, 13,
1499, doi:10.3390/ani13091499.
62. Maleki, M.; Noorimotlagh, Z.; Mirzaee, S.A.; Jaafarzadeh, N.; Martinez, S.S.; Rahim, F.; Kaffashian, M.
An updated systematic review on the maternal exposure to environmental pesticides and involved
mechanisms of autism spectrum disorder (ASD) progression risk in children. Rev. Environ. Health 2023,
38, 727–740, doi:10.1515/reveh-2022-0092.
63. Polańska, K.; Jurewicz, J.; Hanke, W. Review of current evidence on the impact of pesticides,
polychlorinated biphenyls and selected metals on attention deficit / hyperactivity disorder in children.
Int. J. Occup. Med. Environ. Health 2013, 26, doi:10.2478/s13382-013-0073-7.
64. Jozkowiak, M.; Piotrowska-Kempisty, H.; Kobylarek, D.; Gorska, N.; Mozdziak, P.; Kempisty, B.;
Rachon, D.; Spaczynski, R.Z. Endocrine Disrupting Chemicals in Polycystic Ovary Syndrome: The
Relevant Role of the Theca and Granulosa Cells in the Pathogenesis of the Ovarian Dysfunction. Cells
2022, 12, 174, doi:10.3390/cells12010174.
65. Muñoz, J.P.; Bleak, T.C.; Calaf, G.M. Glyphosate and the key characteristics of an endocrine disruptor:
A review. Chemosphere 2021, 270, 128619, doi:10.1016/j.chemosphere.2020.128619.
66. Vianna-Jorge, R.; Perini, J. A., Rondinelli, E.; Suarez-Kurtz, G. CYP2C9 genotypes and the
pharmacokinetics of tenoxicam in Brazilians*1. Clin. Pharmacol. Ther. 2004, 76, 18–26,
doi:10.1016/j.clpt.2004.03.002.
67. Perini, J.; Struchiner, C.; Silva-Assunção, E.; Santana, I.; Rangel, F.; Ojopi, E.; Dias-Neto, E.; Suarez-
Kurtz, G. Pharmacogenetics of Warfarin: Development of a Dosing Algorithm for Brazilian Patients.
Clin. Pharmacol. Ther. 2008, 84, 722–728, doi:10.1038/clpt.2008.166.
68. Matsuzaki, R.; Gunnigle, E.; Geissen, V.; Clarke, G.; Nagpal, J.; Cryan, J.F. Pesticide exposure and the
microbiota-gut-brain axis. ISME J. 2023, 17, 1153–1166, doi:10.1038/s41396-023-01450-9.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those
of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s)
disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or
products referred to in the content.
Preprints.org (www.preprints.org) | NOT PEER-REVIEWED | Posted: 31 July 2024 doi:10.20944/preprints202407.2592.v1
