Investigation on the interaction of pyrethroid pesticides to estrogen receptor alpha through computational and experimental methods.

Pyrethroid insecticides are a group of widely used bio-mimetic synthetic pesticides. However, recent studies reported that they could have an accumulation effect in human which may cause series of health problems. Estrogen receptors (ER) are a class of nuclear receptors that are vital in proper physiological behavior of estrogens. To investigate the reproductive toxicity of pyrethroids, homology modeling, molecular docking, molecular dynamic simulations (MDs) were conducted to explore the interaction between pyrethroids and ERα from atomic scale. The human ERα (2YJA) was selected as a template protein for homology modeling. Then eight typical pyrethroids and positive control estradiol were docked to the modeled protein. The highest scoring bifenthrin and the lowest scoring permethrin were chosen for in-depth analysis. MDs showed that the complex formed by permethrin with ERα had a lower RMSD value and binding free energies compared to bifenthrin. Based on these results from microscopic dimension, exposure experiments were implemented to validate the primary conclusions. VTG concentrations in male zebrafish's blood were significantly higher under permethrin exposure than bifenthrin, suggesting a stronger estrogenic activity and binding propensity. In this regard, the structural characteristics of molecules were analyzed, expecting to provide theoretical references for subsequent drug design and rational drug application.

Investigation of steatosis profiles induced by pesticides using liver organ-on-chip model and omics analysis.

Several studies have reported a correlation between pesticides exposure and metabolic disorders. Dichlorodiphenyltrichloroethane (DDT) and permethrin (PMT), two pesticides highly prevalent in the environment, have been associated to dysregulation of liver lipids and glucose metabolisms and non-alcoholic fatty liver disease (NAFLD). However, the effects of DDT/PMT mixtures and mechanisms mediating their action remain unclear. Here, we used multi-omic to investigate the liver damage induced by DDT, PMT and their mixture in rat liver organ-on-chip. Organ-on-chip allow the reproduction of in vivo-like micro-environment. Two concentrations, 15 and 150 µM, were used to expose the hepatocytes for 24 h under perfusion. The transcriptome and metabolome analysis suggested a dose-dependent effect for all conditions, with a profile close to control for pesticides low-doses. The comparison between control and high-doses detected 266/24, 256/24 and 1349/30 genes/metabolites differentially expressed for DDT150, PMT150 and Mix150 (DDT150/PMT150). Transcriptome modulation reflected liver inflammation, steatosis, necrosis, PPAR signaling and fatty acid metabolism. The metabolome analysis highlighted common signature of three treatments including lipid and carbohydrates production, and a decrease in amino acids and krebs cycle intermediates. Our study illustrates the potential of organ-on-chip coupled to multi-omics for toxicological studies and provides new tools for chemical risk assessment.

Biomonitoring and evaluation of permethrin uptake in forestry workers using permethrin-treated tick-proof pants.

We conducted a randomized case-control trial to analyze uptake of the insecticide/arcaricide permethrin in wearers of permethrin-impregnated and non-impregnated pants in German forestry. Eighty-two male workers were each equipped for a 16-week period with permethrin-treated (test group) or with non-treated work pants (control group). Pants with or without lining to protect against cuts, obtained from two different distributors, were worn in each group. Urinary permethrin metabolite levels were measured by GC-MS/MS before, during and after wearing of the pants. Permethrin uptake was calculated using additional questionnaire data. In the control group, metabolite levels in the range of environmental background exposure (median: ~0.5 µg/l) were measured. Subjects wearing impregnated pants showed consistently significantly higher exposure levels even before the first use of the pants with a maximum after 1 week of wearing the pants (median: ~12.5 µg/l). Significant differences in internal exposure were found depending on which of the distributors the pants came from. Metabolite levels decreased probably due to permethrin losses associated with laundering the pants. Calculated permethrin uptake is below the value corresponding to the WHO-proposed acceptable daily intake. Based on our data, a marginally increased cancer risk compared with the general population cannot be excluded when wearing impregnated pants over a working-lifetime period.

Evaluation of the stereoselective biotransformation of permethrin in human liver microsomes: contributions of cytochrome P450 monooxygenases to the formation of estrogenic metabolites.

Permethrin (PM) is a pyrethroid insecticide that exists as 4 enantiomers. Biotransformation of PM to estrogen receptor agonists (3-phenoxybenzyl alcohol (PBOH) and 3-(4'-hydroxyphenoxy)-benzyl alcohol (3,4 PBOH)) has been shown to be stereoselective in other vertebrate species. This study evaluated the biotransformation of PM enantiomers in human liver microsomes and with recombinant CYP3A4 and CYP2C19. PBOH and 3,4 PBOH were the only metabolites detected from in vitro incubations including each of the 4 enantiomers of PM with 1R-trans PM having the most efficient NADPH-catalyzed biotransformation to both metabolites. Coincubation with the CYP inhibitor ketoconazole and time course experiments with liver microsomes and recombinant CYP2C19 and CYP3A4 indicated CYP-catalyzed stereoselective cleavage of the ester followed by 4-hydoxylation to 3,4' PBOH. These data indicate potential dispositional differences may occur with PM enantiomers and a shift in putative molecular targets. While cleavage of pyrethroid esters lead to detoxification of the acute neurological effects, formation of the benzyl alcohol and hydroxylated metabolite may lead to estrogenic responses, since each of these metabolites are estrogen receptor ligands.

Malaria prevention reduces in-hospital mortality among severely ill tuberculosis patients: a three-step intervention in Bissau, Guinea-Bissau.

BACKGROUND: Malaria and Tuberculosis (TB) are important causes of morbidity and mortality in Africa. Malaria prevention reduces mortality among HIV patients, pregnant women and children, but its role in TB patients is not clear. In the TB National Reference Center in Guinea-Bissau, admitted patients are in severe clinical conditions and mortality during the rainy season is high. We performed a three-step malaria prevention program to reduce mortality in TB patients during the rainy season. METHODS: Since 2005 Permethrin treated bed nets were given to every patient. Since 2006 environmental prevention with permethrin derivates was performed both indoor and outdoor during the rainy season. In 2007 cotrimoxazole prophylaxis was added during the rainy season. Care was without charge; health education on malaria prevention was performed weekly. Primary outcomes were death, discharge, drop-out. RESULTS: 427, 346, 549 patients were admitted in 2005, 2006, 2007, respectively. Mortality dropped from 26.46% in 2005 to 18.76% in 2007 (p-value 0.003), due to the significant reduction in rainy season mortality (death/discharge ratio: 0.79, 0.55 and 0.26 in 2005, 2006 and 2007 respectively; p-value 0.001) while dry season mortality remained constant (0.39, 0.37 and 0.32; p-value 0.647). Costs of malaria prevention were limited: 2€/person. No drop-outs were observed. Health education attendance was 96-99%. CONCLUSIONS: Malaria prevention in African tertiary care hospitals seems feasible with limited costs. Vector control, personal protection and cotrimoxazole prophylaxis seem to reduce mortality in severely ill TB patients. Prospective randomized trials are needed to confirm our findings in similar settings.

Stereoselective biotransformation of permethrin to estrogenic metabolites in fish.

This study investigated the stereoselective biotransformation and resulting estrogenic activity of the pyrethroid insecticide, permethrin (PM). Results of both in vivo (male Japanese medaka, vitellogenin (VTG) protein in plasma) and in vitro (primary rainbow trout hepatocyte VTG-mRNA expression) assays indicated stereoselective estrogenic activity of PM. 1S-cis-PM was observed to have significantly higher activity (P ≤ 0.05) than the 1R-cis enantiomer in both in vivo and in vitro evaluations. All enantiomers of PM were oxidized to a 4'-hydoxy PM (4OH PM) metabolite and underwent esterase cleavage to 3-phenoxybenzyl alcohol (3-PBOH) and 3-(4'-hydroxyphenoxy)-benzyl alcohol) (3,4'-PBOH). Racemic 4OH PM as well as 3-PBOH, and 3,4'-PBOH possessed significant (P ≤ 0.05) estrogenicity. 1S-trans-PM underwent esterase cleavage more extensively than the corresponding 1R-trans-PM. Inhibition studies with ketoconazole confirmed cytochrome P450-catalyzed hydroxylation as well as esterase cleavage of PM for all stereoisomers. These studies indicated stereoselectivity in the estrogenic activity of PM resulting from stereoselective biotransformation of the parent compound to more estrogenic metabolites.

Influence of black carbon and chemical planarity on bioavailability of sediment-associated contaminants.

Black carbon (BC) and chemical properties may play a significant role in defining the bioavailability of hydrophobic organic compounds (HOCs) in sediment. In the current study, bioavailability of four HOCs with differing planarity was determined in sediments amended with two types of BC (soot and charcoal) at different concentrations by matrix solid-phase microextraction (matrix-SPME) and bioaccumulation testing using the freshwater oligochaete Lumbriculus variegatus. Furthermore, the applicability of the matrix-SPME method to bioavailability estimation in BC-amended sediment was tested. The charcoal treatment significantly reduced the bioaccumulation of the planar compounds (3,3',4,4'-tetrachlorobiphenyl and benzo[a]pyrene) in L. variegatus, and the matrix-SPME method showed a similar trend as contaminant bioaccumulation in L. variegatus. Conversely, manipulation of sediment with soot had no effect or slightly increased bioavailability of the planar compounds in both bioaccumulation and matrix-SPME tests. Little if any affect was noted in bioavailability of the nonplanar compounds (2,2',4,4',5,5'-hexachlorobiphenyl and permethrin) with the soot and charcoal amendments. Results showed that the role of BC in defining bioavailability of HOCs depends not only on the type and concentrations of BC present, but also the planarity of the HOCs.

Estrogenic activities of two synthetic pyrethroids and their metabolites.

Synthetic pyrethroids (SPs) are among the most common pesticides in current use, and so far, several SPs have been assessed for their potential estrogenicities by various methods. Previous studies have shown that the estrogenicities partly come from their metabolites. Although considerable information is available with respect to the metabolism and environmental degradation of SPs, little is known about the estrogenicities of the metabolites. In this study, permethrin (PM) and beta-cypermethrin (CP), as well as their metabolites (3-phenoxybenzoic alcohol (PBCOH), 3-phenoxybenzaldehyde (PBCHO) and 3-phenoxybenzoic acid (PBCOOH) were evaluated for their estrogenic activities in the MCF-7 human breast carcinoma cell line. In the MCF-7 cell proliferation assay, PM and CP exhibited significant estrogenic activities at 10(-7) mol/L, comparable to 17beta-estradiol (E2) of 10(-9) mol/L, with the relative proliferative effect ratios of 55.4% and 56.3%, respectively. The real-time quantitative polymerase chain reaction (qRT-PCR) results confirmed the estrogenicities of PM and CP with significant alteration of pS2 and ERalpha mRNA levels observed at 10(-6) mol/L. For the three major metabolites, PBCOH and PBCOOH exhibited estrogenic activities in all assays, while no significant estrogenic responses was observed for PBCHO compared to the vehicle control. In particular, PBCOH had even slightly stronger estrogenic activity than its parent compounds, indicating that metabolism may be one of the reasons for the estrogenicities of the SPs. Given the widespread use of SPs, the toxicological effects of parent compounds and their metabolites should be taken into consideration in the risk assessment of SPs.

Antiandrogenic activity of pyrethroid pesticides and their metabolite in reporter gene assay.

Many pesticides possess hormonal activity and have thus been classified as endocrine disruptors. Pyrethroids are commonly used pesticides worldwide, but little has been done to characterize their antiandrogenic activity potential. We tested three frequently encountered pyrethroids (fenvalerate, cypermethrin, permethrin) and their metabolite 3-phenoxybenzoic acid (3-PBA) for antiandrogenic and androgenic activity using a human androgen receptor (AR) mediated luciferase reporter gene assay in CV-1 African green monkey kidney cell. The assay displayed appropriate response to the known AR agonist 5alpha-dihydrotestosterone and AR antagonist nilutamide and flutamide. At 0.1mM, all the three tested pyrethroids significantly suppressed the luciferase expression. Further, their metabolite 3-PBA also showed antagonist activity. None of the test chemicals showed androgenic activity. Through the antiandrogenic pathways, exposure to certain pyrethroids may contribute to the damage of reproductive system. In conclusion, pyrethroid pesticides can act as antiandrogen in vitro, and metabolizing to 3-PBA cannot eliminate the antagonist activity. This result provides useful information for risk assessment of pyrethroid pesticides.

Estrogenicity of pyrethroid insecticide metabolites.

There is concern that insecticides are able to mimic the action of 17beta-estradiol by interaction with the human estrogen receptor. Pyrethroids are commonly used insecticides and several have been assessed for potential endocrine disrupting activity by various methods. It has been noted that some metabolites of pyrethroids, in particular, permethrin and cypermethrin, have chemical structures that are more likely to interact with the cellular estrogen receptor than the parent pyrethroid. For this study permethrin and cypermethrin metabolites 3-(4-hydroxy-3-phenoxy)benzyl alcohol, 3-(4-hydroxy-3-phenoxy)benzoic acid, and N-3-(phenoxybenzoyl)glycine were synthesised, and together with the commercially available 3-phenoxybenzyl alcohol, 3-phenoxybenzaldehyde, and 3-phenoxybenzoic acid, were studied in a recombinant yeast assay expressing human estrogen receptors (YES). Three metabolites, 3-phenoxybenzyl alcohol, 3-(4-hydroxy-3-phenoxy)benzyl alcohol, and 3-phenoxybenzaldehyde, showed estrogenic activity of approximately 10(5) less than that of 17beta-estradiol. No activity was observed in the yeast assay for 3-phenoxybenzoic acid, 3-(4-hydroxy-3-phenoxy)benzoic acid, and N-3-(phenoxybenzoyl)glycine. The results from this study show that pyrethroid metabolites are capable of interacting with the human estrogen receptor, and so might present a risk to human health and environmental well being. The impact would be expected to be small, but still add to the overall environmental xenoestrogen load.

Human exposure to mosquito-control pesticides--Mississippi, North Carolina, and Virginia, 2002 and 2003.

Public health officials weigh the risk for mosquito-borne diseases against the risk for human exposure to pesticides sprayed to control mosquitoes. Response to outbreaks of mosquito-borne diseases has focused on vector control through habitat reduction and application of pesticides that kill mosquito larvae. However, in certain situations, public health officials control adult mosquito populations by spraying ultra-low volume (ULV) (<3 fluid ounces per acre [oz/acre]) mosquito-control (MC) pesticides, such as naled, permethrin, and d-phenothrin. These ULV applications generate aerosols of fine droplets of pesticides that stay aloft and kill mosquitoes on contact while minimizing the risk for exposure to persons, wildlife, and the environment. This report summarizes the results of studies in Mississippi, North Carolina, and Virginia that assessed human exposure to ULV naled, permethrin, and d-phenothrin used in emergency, large-scale MC activities. The findings indicated ULV application in MC activities did not result in substantial pesticide exposure to humans; however, public health interventions should focus on the reduction of home and workplace exposure to pesticides.

Genotoxicity studies on permethrin, DEET and diazinon in primary human nasal mucosal cells.

Possible genotoxic effects exerted by three widely used pesticides, permethrin, N,N-diethyl-m-toluamide (DEET) and diazinon, in primary human nasal mucosal cells were investigated. Primary nasal mucosa cells were prepared from tissue biopsies taken from 21 patients who underwent nasal surgery. Cells were exposed to 0.5-1.0 mM concentrations of permethrin, DEET and diazinon for 60 min. Genotoxic effects were detected by the alkaline microgel electrophoresis assay ("comet assay"). Within the concentration range, no significant cytotoxic effects were observed, but all three tested pesticides showed a significant genotoxic response that was concentration dependent. More pronounced genotoxic effects were observed in mucosal cells from the middle turbinate than in the inferior turbinate. The results provide some evidence for the potential carcinogenicity of these agents to human nasal mucosal cells. This should be further investigated.