InTechOpen uses cookies to offer you the best online experience. By continuing to use our site, you agree to our Privacy Policy.

Agricultural and Biological Sciences » "Toxicity and Hazard of Agrochemicals", book edited by Marcelo L. Larramendy and Sonia Soloneski, ISBN 978-953-51-2145-9, Published: July 22, 2015 under CC BY 3.0 license. © The Author(s).

# Environmental Pesticides and Heavy Metals — Role in Breast Cancer

By David R. Wallace
DOI: 10.5772/60779

Article top

## Overview

Figure 1. Chemical construction comparison between β-estradiol and pesticides with known or suspected estrogenic activity. Figures all obtained from ChemSpider [www.chemspider.com]

Figure 2. General schematic of the effects of estrogen, xenoestrogens [XenoE2], metalloestrogens [MetalloE2], and phytoestrogens [PhytoE2] on cellular function and the resulting physiological/pathological outcomes. Modified from Darbe, 2014 [97] [PhytoE2 = isoflavones, etc.; XenoE2 = pesticides, etc.; MetalloE2 = heavy metals like cadmium, etc.].

# Environmental Pesticides and Heavy Metals — Role in Breast Cancer

David R. Wallace1

## 1. Introduction

Agriculture chemicals, otherwise referred to as “agrochemicals, ” are a large family of chemicals that cover many pest issues associated with farming. For nearly 5, 000 years, crops have been protected by some form of “pesticide.” Some of the earliest recorded use of “pesticides” was nearly 5, 000 years ago and involved the use of sulfur dusting in the area of modern-day Iraq and surrounding lands. Ancient Sanskrit hymns (Rigveda) allude to the use of various plant-derived compounds, some of which are poisonous, that can be applied to crops, killing insects yet leaving the crops intact – the first insecticides [1]. There was little advancement in the field of pesticides for thousands of years. In the 1400s, the use of chemicals was tried by farmers to kill various crop-related insects. In most cases, the active ingredient of these chemicals was rooted in the actions of heavy metals (arsenic, mercury, and lead). Two hundred years later, the alkaloid, nicotine, was being investigated as a potential agent to eliminate insects from crops. The utility of nicotine prompted additional investigation into the use of natural products as insecticide/pesticide agents. Interestingly, two of the compounds which were developed, pyrethrum and rotenone, are still used today experimentally for various research-related purposes. The use of arsenic-based compounds prevailed for hundreds of years until the mid-1900s as not only agents for pest prevention, but also for poisonings [2]. A shift in the pesticide industry began to occur in the early- to mid-1900s. This began with the identification of dichlorodiphenyltrichloroethane (DDT) as a potent insecticide. DDT was originally developed 75 years earlier, but it was not until nearly 1940 when Dr. Paul Müller discovered that DDT was a very effective insecticide (later being awarded the Nobel Prize in Physiology or Medicine in 1948 for this discovery). Pesticides that contained chlorine were initially referred to as “organochlorines, ” with DDT being considered the prototype organochlorine, which many subsequent agents were based on. Very quickly, considering the timeline of pesticide development and use over thousands of years, organochlorine pesticides were banned in the United States and replaced by newer pesticide derivatives. By substituting a phosphate for chlorine, the “organophosphate” class of pesticides was developed. In addition, the carbamates class was also developed and the organophosphates and carbamates effectively replaced organochlorine pesticides by 1975. Since then, pyrethrin compounds have become the dominant insecticide. Herbicides became common in the 1960s, led by “triazine and other nitrogen-based compounds, carboxylic acids such as 2, 4-dichlorophenoxyacetic acid, and glyphosate” [2].

### 2.2. Pesticides and the link to cancer

While agriculture has traditionally been tied to pesticide-related illnesses, over half of the commonly used pesticides are linked to cancer. The true burden of environmentally induced cancer is greatly underestimated. Chemicals can trigger cancer in a variety of ways, including disrupting hormones, damaging DNA, inflaming tissues, and turning genes on or off. Many pesticides are known to cause cancer, and virtually everyone in the United States is exposed to them on a daily basis. In animal studies, many pesticides are carcinogenic (e.g., organochlorine, creosote, and sulfallate), while others (notably, the organochlorine agents DDT, chlordane, and lindane) are tumor promoters. Some contaminants, such as arsenical compounds, in commercial pesticide formulations also may pose a carcinogenic risk. In humans, arsenic compounds and certain insecticides used occupationally have been classified as carcinogens with organochlorine insecticides being linked with cancers of the lung and breast [47]. With the development of new technologies, the casual association of pesticides with cancer has been strengthened. Most of the human studies have involved retrospective studies examining the incidence of particular cancers like breast cancer and their exposure to pesticides. Now, there is a great deal of research and focus on the identification of biomarkers that can be used to identify these associations between pesticides and cancer. One technology that may be invaluable for biomarker development will be the use of toxicoproteomic-based data [48]. Regardless of the direction that the technology takes us in the future, there is a significant amount of work that still needs to be done to more fully understand how pesticides can be affecting the development of breast cancer as well as other chronic human disorders [49].

Farmers in many countries, including the United States, have lower overall death rates and cancer rates than the general population. Although death rates may be lower, in-depth analysis of the incidence of specific diseases (cancer) indicate that there may be a higher rate of certain cancers among farmers and agricultural workers. Additional work attempting to correlate the types of disease with the chemical exposure has not been unequivocal and there have been many conflicting reports either supporting or refuting the cancer–pesticide link. Similar to what was discussed earlier, the developing young, from fetus to mid-teen, are some of the highest risk groups for disease resulting from pesticide exposure. Young females who may be exposed to an endocrine disrupting agent (such as DDT) have been reported to have a higher incidence of breast cancer compared to populations which were not exposed. Interestingly, this same risk exists if the parents are exposed prior to conception, suggesting that either the pesticide, or changes in the maternal system, can be passed onto the fetus. Collectively, these rural populations of agricultural works and their families tend to be exposed at a higher rate and at higher concentrations than the general population, and as such may experience a higher incidence of particular cancers (breast) than the general population [6].

Yet, to say that there is a significant correlation between pesticide exposure and breast cancer incidence would be an overstatement. Retrospective studies have yielded conflicting results. Most of the studies recognize shortcomings, such as small population size, difficulty in assessing pesticide exposure, and correlating blood pesticide levels to the progression of breast cancer. Pesticides that had widespread use, and were widely popular and commercially available, DDT, DDE, and dieldrin, are the best examined. Since many of these pesticides have exhibited severe health-related adverse effects, older pesticides have been banned, or their use restricted. Yet, due to bioaccumulation, persistence in the environment, and the need to dispose of older stockpiles, they remain an environmental and health concern. As the conflicting evidence has come forward, it appears that one factor that may be important for the toxicity to DDT/DDE/dieldrin is ethnic background. This suggests that genetic polymorphisms between ethnic groups may predispose individuals to cancer risk [50]. A post hoc meta-analysis by Ingber et al. [51] concluded that there was no definite correlation between DDT/DDE exposure and breast cancer. This was done via a PubMed and Web of Science search of nearly 500 cases. Although there were slight elevations in the levels of DDT and the incidence of breast cancer, none of their correlations were statistically significant. Of course, the analysis may have missed some ethnic groups or other populations where a positive correlation existed, but offers a strong conclusion that these compounds alone may not be enough to promote breast cancer development. In an Indian population from Jaipur, women with higher levels of DDT and its metabolites, dieldrin and heptachlor, were correlated with the incidence of breast cancer [52]. A study examining woman of Caribbean descent aimed to correlate pesticide exposure with the incidence of both prostate and breast cancer. Due to relatively high organochlorine pesticide use on the island of Martinique, and what appeared to be an elevated frequency of prostate and breast cancers, the study intended to draw a correlation between pesticide exposure and cancer. They report that there is a positive correlation in the exposure to pesticides and the incidence of breast cancer [53]. Contrary to these studies, research into the causal relationship between pesticides and breast cancer in the United States has not been as convincing and in some instances mixed [50]. An east coast study examined a population of women from Long Island, New York, and found that there was some evidence for a positive correlation between exposure and cancer. It was not a strong correlation, and involved other factors, but would warrant further investigation [54]. Yet a west coast study, the “California Teachers Study” cohort showed no correlation between pesticide exposure and the incidence of breast cancer [55]. Clearly, there is a need to correlate all of the different factors involved in the pathogenesis of breast cancer with the exposure to suspected carcinogens.

It has been known for some time that select pesticides interfered with the action of estrogen. This term was loosely referred to as an “Endocrine Disruptor.” This category has since been expanded to include many items such as detergents, disinfectants, plastics, and an increasing number of pesticides. There are three main actions of an endocrine disruptor. First, the pesticide can mimic the actions of estrogen (or testosterone), thereby causing an increase in estrogen-related physiological responses. Second, the pesticide can act like an antagonist and block the actions of estrogen at its receptors. This will prevent the normal physiological responses associated with estrogen stimulation of its receptor. Third, the pesticide may have a broader effect and interfere with the synthesis, transport, metabolism, or elimination of estrogen. This may have a variable effect with either an increase or decrease in estrogenic effects being observed. Regardless, the normal homeostasis of the system will be disrupted.

Of the broad classes of pesticide – the organochlorine class has been the most extensively studied and has shown to be the most potent as an endocrine disruptor. In the 1990s, studies were done in cell culture model systems which demonstrated that organochlorine pesticides were more potent at activating the estrogen receptor, whereas organophosphorus pesticides were relatively ineffective at modifying estrogen receptor activity [56]. Although the focus of this chapter is breast cancer, other sites of estrogen activity, such as the uterus, also demonstrated that organochlorine pesticides can stimulate receptors on the uterus leading to the development of uterine leiomyoma [57]. The preponderance of data has focused on the ability of these compounds to act directly on estrogen receptors. Initial screening, if originally negative for estrogen receptor activity, does not necessarily mean that organochlorine pesticides are devoid of cancer-stimulating properties. There are indirect mechanisms by which pesticides can influence the proliferation and function of breast tissue. One possible indirect mechanism can be through the binding to tubulin and arrest of the G2/M cycle [58]. A recent study [59] has substantiated the ability of pesticides to influence cellular proliferation. Ventura et al. [59] demonstrated that chlorpyrifos induces a redox imbalance altering the antioxidant defense system and inhibition of cellular proliferation of ERK1/2 phosphorylation. They concluded that the effect on ERK1/2 phosphorylation was not a direct effect but an indirect effect as a result of the changes in the redox state of the cell. Also, most exposures will involve mixtures of compounds. Recent work has begun to explore these effects of mixtures on proliferation of breast tissue. In addition to direct effects of the constituents on the estrogen receptor, the mixtures may also interfere with cellular proliferation [60], inhibition of androgen activity [60], and the upregulation of protein kinase genes associated with tumor development [61]. Not all effects on breast cancer cells are mediated by the genomic actions of estrogen receptors. Activation of CaMKIV pathways is structure-dependent, with estrogen being the most potent activator [62]. Similar results were observed with the activation of PI3-K, MAPK, and PKC. Select phytoestrogens such as resveratrol displayed minimal activity [62]. This would suggest that there is a structural requirement for non-estrogen compounds to elicit effects on intracellular kinase systems. Collectively, the summation of these effects would have a detrimental on cellular function.

In addition to screening for estrogen receptor activity, other methodologies are being developed to further understand the actions of these endocrine disruptors. Estradiol hydroxylase activity has been examined by numerous investigators as a potential predictor of carcinogenic activity. In particular, estradiol 2-hydroxylase has received much attention as a potential predictor. Yet, the results have not been clear and the interpretation of these findings has been complicated, leading to the conclusion that estradiol 2-hyroxylase is not the best predictor of carcinogenic activity [70]. The enzyme “aromatase” is vital for the conversion of testosterone to β-estradiol. A logical extension would be to examine pesticides for their ability to inhibit aromatase (or CYP19 aromatase) activity similar to estrogen [71]. Under physiological conditions, estrogen would negatively feedback onto the aromatase enzyme reducing activity and reducing the amount of estrogen being synthesized from testosterone. Similar to the results observed in the estrogen receptor assays, fungicides such as prochloraz and imazalil inhibited aromatase activity to a greater extent than 4-hydroxyandrostendione. Other pesticides did inhibit aromatase activity but at significantly reduced efficacy and potency. Nearly 33% of the compounds tested exhibited some form of aromatase inhibition [72]. More recently, these findings were substantiated by Sanderson et al. [73]. They report that many of the compounds which may exhibit weak aromatase inhibition did so at concentrations that were cytotoxic to their cell system, R295R cells [73]. They did describe similar effects with fungicides and their effect on inhibiting aromatase activity, but it was suggested that these effects may not be through direct inhibition of aromatase activity, but through inhibition of phosphodiesterase activity [73]. Changes in human CYP19 aromatase expression may then lead to a predisposition to cancer development. One study examined a polymorphism in the CYP19 gene in a Greek population and linked this polymorphism, the population exposure to pesticides, and the incidence of breast cancer [74]. This study did not find a strong association between the short tandem repeat polymorphism, pesticide exposure, and breast cancer development. It is obvious that there are many avenues that can be traveled for the development of breast cancer and that many of the compounds that were viewed to be promoters of tumor formation may in fact function through multiple pathways, and with mixtures of agents, the potential interactions may go up exponentially. A recent report by Sitgaard-Kjeldsen et al. [75] substantiated these conclusions. Using mixtures of pesticides, they report that the observed outcomes are mediated by alterations in ERα, ERβ, androgen receptors, and aromatase [75]. Where understanding the effects of single compounds is important, more fully elucidating the combined actions of pesticides on the development of breast cancer is tantamount to completely understanding the actions of these compounds.

## 4. Combination of metalloestrogens and pesticides

There has been extensive work done examining the individual actions of heavy metals or pesticides in the environment. There has been virtually no work done examining the combined effects of heavy metals and pesticides in the environment. This relative dearth of information has left a significant void in our understanding of the actions of these compounds and their potential role in the development of breast cancer. Many of the compounds discussed in this chapter have demonstrated the ability to interact with many of the pathways associated with tumorigenesis. Interactions with the function of a variety of caspases, Akt/mTOR pathway, p53, ERK1 and 2 signaling pathways, etc. have created a clear need for extensive work in these areas. In many instances, there are no clear direct interactions, but may in fact act through various steps in one or more of these cell cycle cascades. It is quite clear that pesticides and heavy metals do coexist in the environment. There have been numerous reports outlining the effects/coexistence of these compounds in various ecosystems and at various levels of the food chain.

Current knowledge is lacking regarding the collective effects of estrogen and all of the various estrogen-like compounds. Only in the last two decades has there been increasing amounts of work examining the estrogenic effects of naturally occurring compounds as well as synthetic compounds such as pesticides and metals. What is unknown is what overall effect all of these agents will have on a person. These effects have been shown as early as following in utero exposure [95] or through prolonged exposure to trace levels of contaminants leading to endometriosis [96]. The basic physiological responses for estrogen are to control secondary sexual characteristics, influence metabolic activity, central nervous system effects, effects on bone turnover, and, through aromatase, interplay with testosterone. Estrogen can exert these effects by a receptor-mediated mechanism – through estrogen receptor alpha [ERα], estrogen receptor beta [ERβ], and through GPR30. Once stimulated, these receptors will work through intracellular mechanisms leading to nongenomic and genomic responses, and with improving technology, we know that estrogen can regulate hundreds of genes – with the majority (~70%) being downregulated [97, 98]. Since complete pathways have not been elucidated for each of the main classes of exogenous estrogenic compound, it is not known or it is unclear whether there are additive, synergistic, or potentiating effects of these compounds when humans are coexposed. This is one of the troubling aspects of our current knowledge. We have little idea of what effects polyexposure will have on a human, and at what level or threshold will we begin to see these effects. An enormous amount of work still needs to be done in these areas to determine the safe exposure levels, not just for the known compounds, but also for combinations of the compounds [99, 100].

Not only are the potential pathways taken by each of the estrogenic compounds highly complex, but the process of carcinogenesis is also very complex. These processes can be highly involved with many steps and processes working in concert to finally yield a malignant transformation [11]. Cancers may be single-cell in origin and with the mutation of a few genes, errors are expressed that lead to errors in replication and/or growth. In addition, these malignant/mutated cells, dependent on the environment (pollution, toxins, etc.) can cause the conversion of otherwise “normal” cells to cancerous [11]. These “gene–environment interactions” (GEI) are broadly defined as interactions between environmental exposures and specific (risk) genotypes. The term GEI refers to the joint influences of genetic and environmental factors on health and disease. Environmental exposures affect gene regulation and/or act as additive risk factors in conjunction with a particular allelic form of a gene (genetic polymorphism), influencing disease initiation and progression. GEI also entails the different effects of a given environmental exposure on individuals and the different effects of a genotype in people with different histories of environmental exposure. For an excellent review of GEI and the mechanism(s) by which GEI can lead to cancer formation, refer to Tabrez et al. [11]. Kiyama et al. [98] reported a comprehensive listing of various genes which are regulated by estrogen and reviewed their activity. They focused on the classical “endocrine disruptors, ” by focusing on their cell signaling. Their studies first examine gene expression profiles, followed by cell signaling responses. The signaling pathways identified could be used as candidate toxicity pathways to monitor and evaluate endocrine disruptor action [98].

In addition to in vitro studies, there have been human studies which have addressed the concerns associated with the combinations of pesticides and heavy metals. There is a considerable burden of in vitro and in vivo immunotoxicity evidence regarding the detrimental actions of pesticides and heavy metals. Yet, as evidence and information mounts, the findings are still far from unequivocal. Between differences in study design, test subjects, data analysis, and model systems used, etc., it has been virtually impossible to develop a clear correlation between these environmental agents and incidence of disease [99, 100]. Making the correlation between low-level exposures in animal studies to the immune system altering effects observed in humans has been difficult. Also, the effects on human health of the synergistic interactions between natural, medical, dietary, and environmental estrogens have not been fully elucidated yet. There are several factors which need to be accounted for when examining the effects of environmental estrogens: 1) immune status (immunocompromised would have larger response) of the individual, 2) gender (females more responsive than males), 3) status of poly-pollutant exposure, and 4) duration of the exposure. Exposure to the metalloestrogen arsenite in utero altered mammary gland development prior puberty [95]. There was an overgrowth leading up to puberty and a densifying of the breast tissue. After puberty, there was a clear upregulation in the density of estrogen receptor-alpha (ERα) due to the increased and altered response of the ERα transcripts [95], which may lead to the increased risk of developing breast cancer. The large “ENDO Study” examined 22 trace elements and found that 19 of the 22 (86%) of the elements (mostly heavy metals) that were examined were not correlated with endometriosis [96]. Yet, the remaining three that did appear to correlate with endometriosis were cadmium, chromium, and copper – 3 metals that are known metalloestrogens. Additional work will need to be done to substantiate these findings and support their conclusions.

## 5. Conclusions

Over the past 20-30 years, it has become increasingly evident that pesticide use can have unwanted physiological effects beyond the acute exposure. Many pesticides have been banned in numerous countries as these health effects have been described. Even with banning some pesticides, there are stockpiles that need to be disposed of, and some are still used frequently in developing nations. Initial toxicology analysis has focused on the toxicity of individual compounds but this method of assessment may significantly underestimate the risk associated with these compounds when found in mixtures. Many health organizations are now calling for retesting of these agents, but with new guidelines for assessing the potential risk to human and animal life [101, 102]. Through a variety of complex mechanisms, many of these agents have been shown to interact at the estrogen receptor, both ERα and ERβ. These effects can occur in the absence of estrogen and can potentiate estrogenic effects in many mammalian tissues, such as breast and uterus. These interactions lead to the hypothesis that particular pesticide agents – such as organochlorine pesticides – may disrupt the natural endocrine function (i.e., “endocrine disruptors”) of the organism. Interference with the reproductive systems of aquatic- and land-based wildlife may lead to dwindling populations of species of fish, shellfish, and mammals, potentially leading to their extinction. In humans, studies to establish the correlation between pesticide exposure and breast cancer has not been clear and absolute. There has been evidence of positive correlation within some ethnic populations, whereas other studies have yielded negative correlations. In most instances, these were retrospective studies and the design, subject inclusion, and the number of subjects has limited the ability to draw strong conclusions. Obviously, the effects of long-term human exposure needs further study with strict guidelines. There also needs to be a strengthening of the toxicity testing of pesticide mixtures to avoid underestimating the potential toxic effects.

Select heavy metals have been shown to have estrogenic properties and have since been referred to as “metalloestrogens.” Of these, cadmium has been the most extensively studied and appears to be the most potent metalloestrogen at stimulating the estrogen receptor. Both the affinity and inhibitory constant at the ERα receptor is approximately 0.5 nM, which is in order with the affinity of estrogen for its receptor. In vitro studies have shown that cadmium, and some other metalloestrogens, can elicit estrogenic effects resulting in elevation of both ERα and ERβ receptor densities, increased expression and activity of intracellular protein kinases, increased density of progesterone receptors, and the increased size of the uterus as well as increased development and proliferation of breast tissues. Collectively, all of these responses in the presence of metalloestrogens led to speculation that metalloestrogens may be correlated to the incidence of breast cancer. Most of the data currently available have been anecdotal, and have involved in vitro assay systems and breast cancer cell lines (such as MCF-7). In these systems, it is clear that cadmium is the most potent of the metalloestrogens at stimulating tumor development through direct actions at the estrogen receptor, as well as intracellular effects which may be indirect but involve the activation of many signaling systems implicated in tumor development. In human studies, these correlations are not as clear. In a few studies, there was a higher concentration of cadmium in the breast tissue compared to controls, but the direct relationship with breast cancer progression was not clear. Other studies have shown no relationship between cadmium exposure and breast cancer development. One shortcoming of the in vitro studies is that they are relatively acute exposure, for a short duration. This makes it extremely difficult to draw comparisons to human exposure. The potential that low-level, decade-long exposure to cadmium (remembering that cadmium will bioaccumulate, so exposure could be just from the body burden) may result in subtle changes which increase the predisposition to breast cancer development. Many groups are now calling for additional investigation into the mechanisms by which metalloestrogens exert their effects. This additional work is critical to better understanding the actions of metalloestrogens at the estrogen receptors (through NMR or X-ray crystallography), and the interaction of metalloestrogens on intracellular signaling systems.

#### Figure 2.

General schematic of the effects of estrogen, xenoestrogens [XenoE2], metalloestrogens [MetalloE2], and phytoestrogens [PhytoE2] on cellular function and the resulting physiological/pathological outcomes. Modified from Darbe, 2014 [97] [PhytoE2 = isoflavones, etc.; XenoE2 = pesticides, etc.; MetalloE2 = heavy metals like cadmium, etc.].

Lastly, to believe that an individual would be exposed to only one agent would be naïve. For example, a smoker working in the pesticide industry would undoubtedly have elevated cadmium levels due to the tobacco smoke, and possible passive exposure to the pesticides through dermal absorption or inhalation. The combination of these compounds may have a great additive effect on the development of breast cancer. Currently, there are few studies that address these types of combination exposures and virtually none involving the human population. Collectively, as our understanding has grown regarding the negative health effects of pesticides and metalloestrogens, a significantly greater number of questions have arisen and has shed light onto obvious gaps in our understand. As much work that has been done in the last 30 years or so, even more work needs to be done in the next decade or two to assist in our understanding of the pathogenesis of diseases like breast cancer. By better understanding the root causes and foundations for breast cancer development, we will be able to focus our toxicological investigations onto those causes. Also, as we improve our understanding, both in the development of breast cancer, and the involvement of pesticides and metalloestrogens, we will be able to develop better biomarkers. It may not be impossible to completely eliminate exposure, but a viable biomarker that will predict with a high degree of reliability will help with therapeutic interventions at a much earlier time, thereby reducing the morbidity and mortality of this disease.

## Acknowledgements

The author wishes to acknowledge the support provided by the Oklahoma State University Center for Health Sciences. Some of this work was funded in part by intramural funds for the study of “Environmental effects of pesticides and heavy metals on breast cancer cells.” In addition, this project was supported in part by the National Institute of General Medical Sciences of the National Institutes of Health through Grant Number 8P20GM103447 which provided summer internships for local undergraduate students.

## References

1 - Rao GVR, Rupela OP, Rao VR, et al. Role of biopesticides in crop protection: present status and future prospects. Ind J Plant Protect 2007; 35(1):1–9.
2 - Ritter SK. Pinpointing Trends In Pesticide Use. Chemical and Engineering News 2009; 87(7) http://cen.acs.org/articles/87/i7/Pinpointing-Trends-Pesticide-Use.html (accessed February 2, 2015)
3 - Pimentel D. Environmental and economic costs of the application of pesticides primarily in the United States. Environment, Development and Sustainability 2005; 7:229-252 DOI 10.1007/s10668-005-7314-2
4 - Pimentel D, Acquay H, Biltonen M, et al. Environmental and economic costs of pesticide use. BioScience 1992; 42:750-760
5 - Cooper J, Dobson H The benefits of pesticides to mankind and the environment. Crop Protect 2007; 26: 1337–1348 DOI 10.1016/j.cropro.2007.03.022
6 - Breast Cancer Fund. Clear Science: Chemicals in Household Products. www.breastcancerfund.org/clear-science (accessed 20 January 2015)
7 - Darbre PD Environmental oestrogens, cosmetics and breast cancer. Best Prac Res Clin Endocrinol 2006; 20:121-143
8 - Choe SY, Kim SJ, Kim HG, et al. Evaluation of estrogenicity of major heavy metals. Sci Total Environ. 2003; 312:15-21 DOI 10.1016/S0048-9697(03)00190-6
9 - Kakkar P, Jaffery FN. Biological markers for metal toxicity. Environ Toxicol Pharmacol 2005; 19:335-349
10 - Helmfrid I, Berglund M, Lofman O, et al. Health effects and exposure to polychlorinated biphenyls (PCBs) and metals in a contaminated community. Environ Int 2012, 44:53-58
11 - Tabrez S, Priyadarshini M, Priyamvada S, et al. Gene-environment interactions in heavy metal and pesticide carcinogenesis, Mutation Research/Genetic Toxicology and Environmental Mutagenesis 2014; 760:1-9 DOI 10.1016/j.mrgentox.2013.11.002
12 - Aronson KJ, Miller AB, Woolcott CG, et al. Breast adipose tissue concentrations of polychlorinated biphenyls and other organochlorines and breast cancer risk. Can Epidemiol Biomarkers & Prevention 2000; 9:55-63 (Downloaded from cebp.aacrjournals.org on February 3, 2015)
13 - Safe S. Endocrine disruptors and human health: is there a problem. Toxicology 2004; 205(1–2):3–10 DOI 10.1016/j.tox.2004.06.032
14 - Murray DW, Lichter SR. Organochlorine residues and breast cancer. New Eng J Med 1998; 338(14):990–991 DOI 10.1056/nejm199804023381411
15 - Calle EE, Frumkin H, Henley SJ, et al. Organochlorines and breast. Cancer Risk CA Can J Clinic 2002; 52:301-309 DOI 10.3322/canjclin.52.5.301
16 - Caserta D, Maranghi L, Mantovani A, et al. Impact of endocrine disruptor chemicals in gynecology. Human Reproduction Update 2008; 14(1):59–72 DOI 10.1093/humupd/dmm025
17 - Roy JR, Chakraborty S, Chakraborty TR. Estrogen-like endocrine disrupting chemicals affecting puberty in humans – a review. Med Sci Mon 2009; 15(6):RA137-145
18 - Patisaul HB, Adewale HB Long-term effects of environmental endocrine disruptors on reproductive physiology and behavior. Front Behavior Neurosci 2009; 3(10):1-18 DOI 10.3389/neuro.08.010.2009
19 - Karalliedde L, Henry J The acute cholinergic syndrome. In: Organophosphates and Health (Karalliedde L, Feldman S, Henry J, Marrs T, eds) 2001; River Edge, World Scientific Publishing ISBN: 978-1-78326-143-7
20 - Environmental Protection Agency [EPA-HQ-OPPT-2004-0109; FRL-8399-7] Final List of Pesticide Active Ingredients and Pesticide Inert Ingredients to be Screened Under the Federal Food, Drug and Cosmetic Act 2009; 74(71):17579-17585
21 - Osborne G, Rudel R, Schwarzman M. Evaluating chemical effects on mammary gland development: A critical need in disease prevention. Reprod Toxicol 2014, http://dx.doi.org/10.1016/j.reprotox.2014.07.077
22 - Clarke BO, Porter NA, Marriott PJ et al. Investigating the levels and trends of organochlorine pesticides and polychlorinated biphenyl in sewage sludge. Environ Int; 2010; 38:323-329
23 - Schaefer WR, Hermann T, Meinhold-Heerlein I et al. Exposure of human endometrium to environmental estrogens, antiandrogens and organochlorine compounds. Fert Steril 2000; 74:558-563
24 - Saoudi A, Frery N, Zeghnoun A, et al. Serum levels of organochlorine pesticides in the French adult population: The French National Nutritional and Health Study (ENNS), 2006-2007. Sci Total Environ 2014; 472:1089-1099
25 - Konar SK, Mullick M. Problems of safe disposal of petroleum products, detergents, heavy metals and pesticides to protect aquatic life. Sci Total Environ 1993; Supplement:989-1000
26 - Becker PR. Concentration of chlorinated hydrocarbons and heavy metals in Alaska arctic marine mammals. Mar Poll Bull 2000; 40(10):819-829
27 - Kumar KS, Sajwan KS, Richardson JP, et al. Contamination profiles of heavy metals, organochlorine pesticides, polycyclic aromatic hydrocarbons and alkylphenols in sediment and oyster collected from marsh/estuarine Savannah GA, USA. Mar Poll Bull 2008; 56:136–162
28 - Maes J, Belpaire C, Goemans G. Spatial variations and temporal trends between 1994 and 2005 in polychlorinated biphenyls, organochlorine pesticides and heavy metals in European eel (Anguilla anguilla L.) in Flanders, Belgium. Environ Poll 2008; 153:223-237 DOI 10.1016/j.envpol.2007.07.021
29 - Yatawara M, Qi S, Owago OJ et al. Organochlorine pesticide and heavy metal residues in some edible biota collected from Quanzhou Bay and Xinghua Bay, Southeast China. J Environ Sci 2010; 22(2):314–320 DOI 10.1016/S1001-0742(09)60110-8
30 - Mazet A, Keck G, Berny P. Concentrations of PCBs, organochlorine pesticides and heavy metals (lead, cadmium, and copper) in fish from the Drôme River: Potential effects on otters (Lutra lutra). Chemosphere 2005; 61:810-816 DOI 10.1016/j.chemosphere.2005.04.056
31 - Sankar TV, Zynudheen AA. Anandan R, et al. Distribution of organochlorine pesticides and heavy metal residues in fish and shellfish from Calicut region, Kerala, India. Chemosphere 2006; 65:583-590 DOI 10.1016/j.chemosphere.2006.02.038
32 - Araújo DFS, Silva AMRB, Lima LLA The concentration of minerals and physicochemical contaminants in conventional and organic vegetables. Food Contr 2014; 44:242-248 DOI 10.1016/j.foodcont.2014.04.005
33 - Mansour SA, Belal MH, Abou-Arab AAK, et al. Monitoring of pesticides and heavy metals in cucumber fruits produced from different farming systems. Chemosphere 2009; 75:601–609 DOI 10.1016/j.chemosphere.2009.01.058
34 - Nasreddine Parent-Massin L. Food contamination by metals and pesticides in the European Union. Should we worry? Toxicol Lett 2002; 127:29-41
35 - Mansour SA, Gad MF, Risk assessment of pesticides and heavy metals contaminants in vegetables: A novel bioassay method using Daphnia magna Straus. Food Chem Toxicol 2010; 48:377–389 DOI 10.1016/j.fct.2009.10.026
36 - Li B, Wu S, Lui C. China pharmaceuticals: Investing in Traditional Chinese Medicine (TCM). Hong Kong, China: Morgan Stanley Research Asia/Pacific; 2009.
37 - Nahin RL, Barnes PM, Stussman BJ et al. Costs of complementary and alternative medicine (CAM) and frequency of visits to CAM practitioners: United States, 2007. National Health Status Report 2009; 18:1–15. http://www.cdc.gov/nchs/data/nhsr/nhsr018.pdf (accessed February 6, 2015)
38 - Varma SP, Boldin BR, Lin PS. The Inhibition of the estrogenic effects of pesticides and environmental chemicals by curcumin and isoflavonoids. Environ Health Persp 1998; 106(12):807812
39 - Ernst E. Toxic heavy metals and undeclared drugs in Asian herbal medicines. Trends Pharmacol Sci 2002; 23:136–139 DOI 10.1016/S0165-6147(00)01972-6
40 - Ernst E, Coon JT. Heavy metals in traditional Chinese medicines: A systematic review. Clin Pharmacol Therapeut 2001; 70:497–504 DOI 10.1067/mcp.2001.120249
41 - Lin CG, Schaider LA, Brabander DJ et al. Pediatric lead exposure from imported Indian spices and cultural powders. Pediatrics 2010; 125:E828–35 DOI 10.1542/peds.2009-1396
42 - Harris ESJ, Cao S, Littlefield BA, et al. Heavy metal and pesticide content in commonly prescribed individual raw Chinese herbal medicines. Sci Total Environ 2011; 409:4297–4305 DOI 10.1016/j.scitotenv.2011.07.032
43 - Liu J, Lu YF, Wu Q et al. Mineral arsenicals in traditional medicines: Orpiment, realgar, and arsenolite. J Pharmacol Exper Therapeut 2008a; 326:363–368 DOI 10.1124/jpet.108.139543
44 - Liu J, Shi JZ, Yu LM, et al. Mercury in traditional medicines: is cinnabar toxicologically similar to common mercurials? Exper Biol Med 2008b; 233:810–817 DOI 10.3181/0712-MR-336
45 - Saper RB, Phillips RS, Sehgal A, et al. Lead, mercury, and arsenic in US- and Indian-manufactured Ayurvedic medicines sold via the Internet. J Am Med Assoc 2008; 300:915–923 DOI 10.1001/jama.300.8.915
46 - US GAO (United States Government Accountability Office). Herbal dietary supplements: examples of deceptive or questionable marketing practices and potentially dangerous advice. Washington, DC, United States: US GAO; 2010; GAO-10-662T http://www.gao.gov/new.items/d10662t.pdf (accessed February 6, 2015)
47 - Dich J, Zahm SH, Hanberg A, Adami HO. Pesticides and cancer. Can Causes Cont 1997; 8:420-443
48 - George J, Shukla Y. Pesticides and cancer: Insights into the toxicoproteomic-based findings. J Proteomics 2011; 74:2719-2722 DOI 10.1016/j.prot.2011.09.024
49 - Mostafalou S, Abdollahi M. Pesticides and human chronic diseases: Evidences, mechanism, and perspectives. Toxicol Appl Pharmacol 2013; 268:157-177 DOI 10.1016/j.taap.2013.01.025
50 - Snedeker SM. Pesticides and breast cancer risk: A review of DDT, DDE and Dieldrin. Environ Health Persp 2001; 109(1):35-47
51 - Ingber SZ, Buser MC, Pohl HR, et al. DDT/DDE and breast cancer: A meta-analysis. Regul Toxicol Pharmacol 2013; 67:421-433 DOI 10.1016/j.yrtph.2013.08.021
52 - Mathur V, Bhatnagar P, Sharma RG, et al. Breast cancer incidence and exposure to pesticides among women originating from Jaipur. Environ Int 2002; 28:331-336
53 - Landau-Ossondo M, Rabia N, Jos-Pelage J, et al. Why pesticides could be a common cause of prostate and breast cancers in the French Caribbean Island, Martinique. An overview on key mechanisms of pesticide-induced cancer. Biomed Pharmacother 2009; 63:383-395 DIO 10.1016/j.biopha.2009.04.043
54 - O’Leary ES, Vena JE, Freudenheim JL, et al. Pesticide exposure and risk of breast cancer: a nested case-control study of residentially stable women living on Long Island. Environ Res 2004; 94:134-144 DOI 10.1016/l.envres.2003.08.001
55 - Reynolds P, Hurley SE, Goldberg DE, et al. Residential proximity to agricultural pesticide use and the incidence of breast cancer in the California Teachers Study cohort. Environ Res 2004; 96:206-218 DOI 10.1016/j.envres.2004.03.001
56 - Bradlow HL, Davis D, Sepkovic DW, et al. Role of the estrogen receptor in the action of organochlorine pesticides on estrogen metabolism in human breast cancer cell lines. Sci Total Environ 1997; 208:9-14
57 - Hodges LC, Bergerson JS, Hunter DS et al. Estrogenic effects of organochlorine pesticides on uterine leiomyoma cells in vitro. Toxicol Sci 2000; 54:355-364
58 - Ventura C, Nunez M, Miret N, et al. Differential mechanisms of action are involved in chlorpyrifos effects in estrogen-dependent or –independent breast cancer cells exposed to low or high concentrations of the pesticide. Toxicol Lett 2012; 213:184-193 DOI 10.1016/j.toxlet.2012.06.017
59 - Ventura C, Venturino A, Miret N, et al. Chlorpyrifos inhibits cell proliferation through ERK1/2 phosphorylation in breast cancer cell lines. Chemosphere 2015; 120:343-350 DOI 10.1016/j.chemosphere.2014.07.088
60 - `Aube M, Larochelle C, Ayotte P. Differential effects of a complex organochlorine mixture on the proliferation of breast cancer cell lines. Environ Res 2011; 111:334-347 DOI 10.1016/j.envres.2011.01.010
61 - Valeron PF, Pestano JJ, Luzardo OP, et al. Differential effects exerted on human mammary epithelial cells by environmentally relevant organochlorine pesticides either individually or in combination. Chem-Biol Interact 2009; 180:485-491 DOI 10.1016/j.cbi.2009.04.010
62 - Li X, Zhang S, Safe S. Activation of kinase pathways in MCF-7 cells by 17β-estradiol and structurally diverse estrogenic compounds. J Steroid Biochem Mol Biol 2006; 98:122-132 DOI 10.1016/j.jsbmb.2005.08.018
63 - Vom Saal FS, Nagel SC, Palanza P et al. Estrogenic pesticides: binding relative to estradiol in MCF-7 cells and effects of exposure during fetal life on subsequent territorial behavior in male mice. Toxicol Lett 1995; 77:343-350
64 - Soto Am, Chung KL, Sonnenschein C. The pesticides endosulfan, Toxaphene, and dieldrin have estrogenic effects on human estrogen-sensitive cells. Human Health Persp 1994; 102:380-383
65 - Grunfeld HT, Bonefeld-Jorgensen EC. Effect of in vitro estrogenic pesticides on human estrogen receptor α and β mRNA levels. Toxicol Lett 2004; 151:467-480 DOI: 10.1016/toxlet.2004.03.021
66 - Hofmeister MV, Bonefeld-Jorgensen EC. Effects of the pesticides prochloraz and methiocarb on human estrogen receptor α and β mRNA levels analyzed by online RT-PCR. Toxicol In Vitro 2004; 18:427-433 DOI10.1016/j.tiv.2003.12.008
67 - Kojima H, Katsura E, Takeuchi S et al. Screening for estrogen and androgen receptor activities in 200 pesticides by in vitro reporter gene assays using Chinese hamster ovary cells. Environ Health Persp 2004; 112(5):524-531 DOI 10.1289/ehp.6649
68 - Lemaire G, Mnif W, Mauvais P et al. Activation of α- and β-estrogen receptors by persistent pesticides in reporter cell lines. Life Sci 2006; 79:1160-1169 DOI 10.1016/j.lfs.2006.03.023
69 - Barker S, Malouitre SDM, Glover HR, et al. Comparison of effects of 4-hydroxy tamoxifen and Trilostane on estrogen-regulated gene expression in MCF-7 cells: Up-regulation of estrogen receptor beta. J Steroid Biochem Mol Biol 2006; 100:141-151 DOI 10.1016/j.jsbmb.2006.04.006
70 - McDougal A, Wilson C, Safe S. Induction of estradiol 2-hydroxylase activity in MCF-7 human breast cancer cells by pesticides and carcinogens. Environ Toxicol Pharmacol 1997; 3:195-199
71 - Andersen HR, Vinggaard AM, Rasmussen TH, et al. Effects of currently used pesticides in assays for estrogenicity, androgenicity, and aromatase activity in vitro. Toxicol Appl Pharmacol 2002; 179:1-12 DOI 10.1006/taap.2001.9347
72 - Vinggaard AM, Hnida C, Breinholt V et al. Screening of selected pesticides for inhibition of CYP19 aromatase activity in vitro. Toxicol In Vitro 2000; 14:227-234
73 - Sanderson JT, Boerma J, Lansbergen GWA et al. Induction and inhibition of aromatase (CYP19) activity by various classes of pesticides in H295R human adrenocortical carcinoma cells. Toxicol Appl Pharmacol 2002; 182:44-54 DOI 10.1006/taap.2002.9420
74 - DialynaI, Tzanakakis G, Dolapsakis G, et al. A tetranucleotide repeat polymorphism in the CYP19 gene and breast cancer susceptibility in a Greek population exposed and not exposed to pesticides. Toxicol Lett 2004; 151:267-271 DOI 10.1016/j.toxlet.2004.01.024
75 - Stigaard-Kjeldsen L, Ghisari M, Bonefeld-Jorgensen EC. Currently used pesticides and their mixtures affect the function of sex hormone receptors and aromatase enzyme activity. Toxicol Appl Pharmacol 2013; 272:453-464 DOI 10.1016/j.taap.2013.06.028
76 - Gimeno-Garcia E, Andreu V, Boluda R. Heavy metals incidence in the application of inorganic fertilizers and pesticides to rice farming soils. Environ Poll 1996; 92(1):19-25
77 - Darbre PD. Review article: Underarm cosmetics and breast cancer. J Appl Toxicol 2003; 23:89-95
78 - Darbre P. Metalloestrogens: An emerging class of inorganic xenoestrogens with potential to add to the estrogenic burden of the human breast. J Appl Toxicol 2005; 26:191-197 DOI: 10.1002/jat.1135
79 - Dyer CA. Heavy metals as endocrine-disrupting chemicals. In: Endocrine-Disrupting Chemicals: From Basic Research to Clinical Practice (ed. Gore AC) 2007; Humana Press Inc. Totowa NJ, 111-133
80 - Georgescu B, Georgescu C, Daraban S, et al. Heavy metals acting as endocrine disruptors. Anim Sci Biotechnol 2011; 44(2):89-93
81 - Byrne C, Divekar SD, Storchan GB, et al. Cadmium – A metallohormone. Toxicol Appl Pharmacol 2009; 238:266-271 DOI: 10.1016/j.taap.2009.03.025
82 - Martin MB, Reiter R, Pham T, et al. Estrogen-like activity of metals in MCF-7 breast cancer cells. Endocrinology 2003; 144:2425-2436
83 - Johnson MD, Kenney N, Stoica A, et al. Cadmium mimics the in vivo effects of estrogen in the uterus and mammary gland. Natur Med 2003; 9:1081-1084
84 - Aquino NB, Sevigny MB, Sabangan J, et al. The role of cadmium and nickel in estrogen receptor signaling and breast cancer: metalloestrogens or not? J Exper Sci Health Part C 2012; 30:189-224 DOI 10.1080/10590501.2012.705159
85 - Byrne C, Divekar SD, Storchan GB et al. Metals and breast cancer. J Mamm Glan Biol Neoplas 2013; 18:63-73 DOI: 10.1007/s10911-013-9273-9
86 - Yu X, Filardo EJ, Shaikh ZA. The membrane estrogen receptor GPR30 mediates cadmium-induced proliferation of breast cancer cells. Toxicol Appl Pharmacol 2010; 245:83–90 DOI 10.1016/j.taap.2010.02.005
87 - Safe S. Cadmium’s disguise dupes the estrogen receptor. Nature Medicine 2003; 9(8):1000-1001
88 - Garcia-Morales P, Saceda M, Kenney N, et al. Effect of cadmium on estrogen-receptor levels and estrogen-induced responses in human breast cancer cells. J Biol Chem 1994; 269:16896-16901
89 - Lortenkamp A. Are cadmium and other heavy metal compounds acting as endocrine disruptors? Metal Ions in Life Sci 2011; 8:305-317
90 - Zhang RY, Liu Y, Pang DW, et al. Spectroscopic and voltametric study on the binding of aluminum (III) to DNA. Japan Soc Anal Chem, 2002; 18:761-766
91 - Ionescu JG, Novotny J, Stejskal V, et al. Increased levels of transition metals in breast cancer tissue. Neuroendocrinol Lett 2006; 27(Suppl 1):36-39
92 - Wu HDI, Chou SY, Chen DR, et al. Differentiation of serum levels of trace elements in normal and malignant breast patients. Biol Trace Element Res 2006; 113:9-18
93 - Silva N, Peiris-John R, Wickremasinghe R. Cadmium a metalloestrogen: are we convinced? J Appl Toxicol 2012; 32:318–332 DOI 10.1002/jat.1771
94 - Stoica A, Katzenellenbogen BS, Martin MB. Activation of estrogen receptor alpha by the heavy metal cadmium. Mol Endocrinol 2000; 14:545-553
95 - Parodi DA, Greenfield M, Evans C, et al. Alteration of mammary gland development and gene expression by in utero exposure to arsenic. Reprod Toxicol 2015; In Press DOI 10.1016/j.reprotox.2014.12.011
96 - Pollack AZ, Buck-Louis GM, Chen Z, et al. Trace elements and endometriosis: The ENDO study. Reprod Toxicol 2013; 42:41-48 DOI 10.1016/j.reprotox.2013.05.009
97 - Darbre PD. Environmental Contaminants: Environmental Estrogens – Hazard Characterization. Encyclopedia of Food Safety 2014; 2:323–331 DOI 10.1016/B978-0-12-378612-8.00196-7
98 - Kiyama R, Zhu Y, Kawaguchi K, et al. Estrogen-responsive genes for environmental studies. Environ Technol Innov 2014; 1-2:16-28 DOI 10.1016/j.eti.2014.09.001
99 - Chighizola C, Meroni PL. The role of environmental estrogens and autoimmunity. Autoimmun Rev 2012; 11:A493-A501 DOI 10.106/j.autrev.2011.11.027
100 - Wessels D, Barr DB, Mendola P. Use of biomarkers to indicate exposure of children to organophosphate pesticides: Implications for a longitudinal study of children’s environmental health. Environ Health Persp 2003; 111(16): 1939-1946 DOI 10.1289/EHP.6179
101 - Pimental D. Environmental and economic costs of application of pesticides primarily in the United States. Environ Dev Sustain 2005; 7:229-252. DOI 10.1007/s10668-005-7314-2
102 - Food and Agriculture Organization of the United Nations (FAO), International Code of Conduct on the Distribution and Use of Pesticides. 2005; ISBN 92-5-105411-8 (accessed Feb 2, 2015)