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1.
Environ Pollut ; 345: 123398, 2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38272163

ABSTRACT

Titanium-incorporated diatoms are promising biomaterials to photodegrade micropollutants such as pharmaceuticals and personal care products (PPCPs). Hydraulic retention time (HRT) is a key parameter for diatom cultivation and the incorporation of titanium into diatom frustules. This study assessed how HRT governs the micro/nanostructures, titania (TiO2) content and distribution, and the photocatalytic activity of titanium-incorporated diatom frustules. We cultivated a diatom strain Stephanodiscus hantzschii using a feed solution containing titanium(IV) in membrane bioreactors (MBRs) at a solids retention time (SRT) of 10 d and staged HRTs from 24 to 12 and to 6 h. The decrease in HRT reduced the porosity of diatom frustules but increased their silicon and titania contents. When the HRT decreased from 24 to 12 and to 6 h, the specific surface areas of the diatom decreased from 37.65 ± 3.19 to 31.53 ± 3.71 and to 18.43 ± 2.69 m2·g-1 frustules, while the titanium (Ti) contents increased from 53 ± 14 to 71 ± 9 and to 85 ± 13 mg Ti·g-1 frustules. The increase in the influent flow rates of the MBRs with decreasing HRTs likely enhanced nutrient diffusion inside the diatom valve pores, facilitating the uptake and incorporation of silicon and titanium. The titanium-incorporated frustules were effective in removing two representative PPCPs, bisphenol A (BPA) and N,N-diethyl-meta-toluamide (DEET), from water. As photocatalytic activity depends on the amount of titanium, decreasing the HRT substantially increased the photocatalytic activity of the titanium-incorporated frustules. In batch tests under ultraviolet light, frustules from the diatom cultivated at HRTs of 24, 12, and 6 h had the pseudo-first-order removal (mainly through photodegradation) rate constants of BPA of 0.376, 0.456, and 0.683 h-1, respectively. Under the same experimental condition, the pseudo-first-order removal rate constants of DEET by the frustules cultivated at HRTs of 24, 12, and 6 h increased from 0.270 to 0.330 and to 0.480 h-1.


Subject(s)
Diatoms , Nanostructures , Diatoms/metabolism , Titanium/chemistry , Silicon/metabolism , DEET/metabolism , Nanostructures/chemistry , Silicon Dioxide/chemistry
2.
Front Public Health ; 11: 1248674, 2023.
Article in English | MEDLINE | ID: mdl-38074723

ABSTRACT

Background: Currently, there is limited research on the specific relationship between N, N-diethyl-m-toluamide (DEET) exposure and the odds of kidney stones. We aimed to investigate the relationship between DEET exposure and the prevalence of kidney stones. Methods: We included 7,567 qualified participants in our research from the 2007-2016 NHANES survey. We carried out three logistic regression models to explore the potential association between DEET exposure and the odds of kidney stones. Spline smoothing with generalized additive models (GAM) was utilized to assess the non-linear relationship and restricted cubic spline (RCS) curves was to determine the dose-response association. Multivariate regression models were used to conduct stratified analysis and sensitivity analysis. Results: Baseline characteristics of study participants presented the distribution of covariables. Regression analysis revealed that the odds of kidney stones were positively associated with the main metabolites of 3-diethyl-carbamoyl benzoic acid (DCBA) (log2) (OR = 1.05, 95% CI 1.02 to 1.08). The fourth quartile of urine DCBA showed a greater risk of kidney stones in the fully adjusted model (OR = 1.36, 95% CI 1.08 to 1.72). Another DEET metabolite of N, N-diethyl-3-hydroxymethylbenzamide (DHMB) was used to confirm the accuracy and stability of the results. The spline smoothing curve represented two main DEET metabolites had similar no-linear relationships and a positive trend with kidney stones proportion. RCS implied that the incidence of kidney stones rose with increasing levels of DEET exposure. High-risk groups on kidney stones were exhibited by stratified analysis under DEET exposure. Conclusion: Our study suggests that DEET exposure is positively associated with odds of kidney stones. Further investigation into the underlying processes of this association is required to guide the prevention and treatment of kidney stones.


Subject(s)
Insect Repellents , Kidney Calculi , Adult , Humans , DEET/metabolism , Insect Repellents/metabolism , Nutrition Surveys , Surveys and Questionnaires , Kidney Calculi/chemically induced , Kidney Calculi/epidemiology
3.
Regul Toxicol Pharmacol ; 145: 105506, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37838349

ABSTRACT

N,N-Diethyl-meta-toluamide (DEET) is widely used as an effective mosquito and tick repellent. DEET is absorbed systemically after applications to skin. Once absorbed, DEET is rapidly metabolized with the predominant metabolite being m-dimethylaminocarbonyl benzoic acid (DBA). DEET and metabolites are predominantly excreted in urine after being absorbed systemically. Exposures to DEET are typically biomonitored via measures of DEET and DBA in urine. In this evaluation, we review available health-based risk assessments and toxicological reference values (TRVs) for DEET and derive Biomonitoring Equivalent (BE) values for interpretation of population biomonitoring data. BEs were derived based on existing TRVs derived by Health Canada, yielding 38 and 23 mg/L DBA in urine for adults and 57 and 34 mg/L DBA in urine in children for the acute oral and intermediate dermal TRVs, respectively. The BEs for unchanged DEET in urine are 21 and 12 mg/L in adults and 4.5 and 2.7 mg/L in children for the acute oral and intermediate dermal TRVs. The BE values derived in this manuscript can serve as a guide to help public health officials and regulators interpret population based DEET biomonitoring data in a public health risk context.


Subject(s)
DEET , Insect Repellents , Adult , Child , Animals , Humans , DEET/metabolism , Biological Monitoring , Skin/metabolism , Benzoic Acid
4.
Front Public Health ; 10: 922005, 2022.
Article in English | MEDLINE | ID: mdl-36267998

ABSTRACT

Although growing evidence suggests that N,N-diethyl-m-toluamide (DEET) has adverse effects on public health, the relationship of DEET with cardiovascular disease (CVD) is still largely unknown. The purpose of this study was, therefore, to evaluate the association between DEET exposure and total and specific CVD among the US adults. In this cross-sectional study, a total of 5,972 participants were selected from the National Health and Nutrition Examination Survey (NHANES) 2007-2014. CVD was defined as a combination of congestive heart failure (CHF), coronary heart disease (CHD), angina, heart attack, or stroke. Logistic regression models were used to evaluate the association between DEET metabolites and the risks of total and specific CVD. Compared to the lowest quartile, 3-(diethylcarbamoyl) benzoic acid (DCBA) in the highest quartile was associated with the increased risks of CVD (odds ratio [OR]: 1.32, 95% CI: 1.03-1.68, P for trend = 0.025) and CHD (OR: 1.57, 95% CI: 1.10-2.25, P for trend = 0.017), after adjustment for potential covariates. Nevertheless, exposure to DCBA was not significantly associated with heart attack, CHF, angina, and stroke. Further studies are required to confirm these findings and identify the underlying mechanisms.


Subject(s)
Cardiovascular Diseases , Myocardial Infarction , Stroke , Humans , Adult , DEET/adverse effects , DEET/metabolism , Cardiovascular Diseases/epidemiology , Cardiovascular Diseases/chemically induced , Nutrition Surveys , Cross-Sectional Studies , Myocardial Infarction/chemically induced , Stroke/chemically induced , Benzoic Acid
5.
An Acad Bras Cienc ; 94(suppl 3): e20211166, 2022.
Article in English | MEDLINE | ID: mdl-36074427

ABSTRACT

DEET is considered the gold standard for insect repellent products. However, it behaves as a strong skin permeant. DEET was encapsulated in Solid Lipid Microparticles (SLM) and characterized in terms of morphology, particle size, cytotoxicity and ex vivo permeation. The particles exhibited micrometric size with a spherical shape. In addition, we developed and validated an analytical method for DEET quantification by high performance liquid chromatography (HPLC), which was selective, linear, precise, accurate and robust. The toxicity test in cell culture of keratinocytes, fibroblasts and macrophages showed that the formulation did not present cytotoxicity. The SLM were able to decrease the skin permeation of DEET in relation to the free active in ethanol with gain in the safe. Microparticles were able to increase the skin retention of DEET, which can contribute to extend the time of repellent action. The results showed that Solid Lipid Microparticles are safe and promising topical formulation to insect bite prevention.


Subject(s)
DEET , Insect Repellents , DEET/chemistry , DEET/metabolism , Insect Repellents/chemistry , Insect Repellents/metabolism , Lipids , Skin , Skin Absorption
6.
Nature ; 597(7874): 126-131, 2021 09.
Article in English | MEDLINE | ID: mdl-34349260

ABSTRACT

Olfactory systems must detect and discriminate amongst an enormous variety of odorants1. To contend with this challenge, diverse species have converged on a common strategy in which odorant identity is encoded through the combinatorial activation of large families of olfactory receptors1-3, thus allowing a finite number of receptors to detect a vast chemical world. Here we offer structural and mechanistic insight into how an individual olfactory receptor can flexibly recognize diverse odorants. We show that the olfactory receptor MhOR5 from the jumping bristletail4 Machilis hrabei assembles as a homotetrameric odorant-gated ion channel with broad chemical tuning. Using cryo-electron microscopy, we elucidated the structure of MhOR5 in multiple gating states, alone and in complex with two of its agonists-the odorant eugenol and the insect repellent DEET. Both ligands are recognized through distributed hydrophobic interactions within the same geometrically simple binding pocket located in the transmembrane region of each subunit, suggesting a structural logic for the promiscuous chemical sensitivity of this receptor. Mutation of individual residues lining the binding pocket predictably altered the sensitivity of MhOR5 to eugenol and DEET and broadly reconfigured the receptor's tuning. Together, our data support a model in which diverse odorants share the same structural determinants for binding, shedding light on the molecular recognition mechanisms that ultimately endow the olfactory system with its immense discriminatory capacity.


Subject(s)
Insect Proteins/chemistry , Insect Proteins/metabolism , Insecta/metabolism , Ion Channel Gating , Odorants/analysis , Receptors, Odorant/chemistry , Receptors, Odorant/metabolism , Animals , Binding Sites , Cell Line , DEET/metabolism , Eugenol/metabolism , Insect Proteins/genetics , Insecta/genetics , Ion Channels/chemistry , Ion Channels/genetics , Ion Channels/metabolism , Models, Molecular , Mutation , Protein Binding , Protein Structure, Quaternary , Receptors, Odorant/genetics , Substrate Specificity
7.
Am J Trop Med Hyg ; 103(2): 812-814, 2020 08.
Article in English | MEDLINE | ID: mdl-32458781

ABSTRACT

N,N-diethyl-meta-toluamide (DEET) is one of the most commonly used insect repellants in the United States, yet the existing literature regarding DEET's potential deleterious impact on humans is mixed and is based mostly on case reports. The primary aim of this study was to address this lack of population-based evidence of the effects of DEET exposure on human health in the United States. Our primary outcome measures were biomarkers related to systemic inflammation (high sensitivity C-reactive protein), immune function (lymphocyte), liver function (aspartate aminotransferase, alanine aminotransferase, and γ-glutamyl transferace), and kidney function (estimated glomerular filtration rate). We analyzed data from the population-based National Health and Nutrition Examination Survey, 2015-2016, and identified 1,205 patients (age 20+ years) who had DEET metabolite levels recorded at or above detection limits. A Pearson correlation was used to assess the relationship between DEET metabolite, and each biomarker found there was no significant correlation. Thus, there is no evidence that DEET exposure has any impact on the biomarkers identified.


Subject(s)
Alanine Transaminase/blood , Aspartate Aminotransferases/blood , C-Reactive Protein/metabolism , DEET/blood , Glomerular Filtration Rate , Insect Repellents/blood , Lymphocyte Count , gamma-Glutamyltransferase/blood , Adult , Aged , Biomarkers , DEET/metabolism , Female , Humans , Insect Repellents/metabolism , Male , Middle Aged , Nutrition Surveys , United States
8.
Med Vet Entomol ; 34(2): 236-239, 2020 06.
Article in English | MEDLINE | ID: mdl-31483877

ABSTRACT

The present study aimed to evaluate the behaviour of larvae of Rhipicephalus microplus exposed to different stimuli. A Y-olfactometer was positioned vertically and R. microplus larvae were exposed to environmental air, CO2 alone, N,N-diethyl-3-methylbenzamide (DEET) alone, and CO2 combined with the repellents DEET and (E)-2-octenal. Tests were also conducted with the olfactometer positioned horizontally; in this case, however, only CO2 was tested. In all tests conducted with the Y-olfactometer positioned vertically, CO2 activated R. microplus larvae even in the presence of DEET and (E)-2-octenal, although activation was lower when these repellents were used. In the absence of CO2 , larval behaviour against DEET was similar to that of the larvae in the control group. In the tests performed with the olfactometer positioned horizontally, the larvae had no significant response to the presence of CO2 . The larvae were not attracted to or repelled by any compound tested in either the vertical or horizontal position of the olfactometer. The lack of horizontal displacement, attraction or repellence may have been a result of the ambush behaviour of this tick species. However, when larvae were exposed to stimuli and the olfactometer was positioned vertically, the interference of attractant and repellent stimuli in larval behaviour was assessed.


Subject(s)
Acaricides/metabolism , Aldehydes/metabolism , Carbon Dioxide/metabolism , DEET/metabolism , Rhipicephalus/physiology , Animals , Insect Repellents/metabolism , Larva/physiology , Olfactometry , Rhipicephalus/growth & development
9.
Chem Senses ; 42(4): 333-341, 2017 05 01.
Article in English | MEDLINE | ID: mdl-28334229

ABSTRACT

In Culex quinquefasciatus, CquiOR91 is the ortholog of 2 larvae-specific odorant receptors (ORs) from Anopheles gambiae (Agam\Or40, previously shown to respond to several odorant ligands including the broad-spectrum repellent N,N-diethyl-3-methylbenzamide, DEET) and Aedes aegypti (Aaeg\Or40). When we cloned full-length CquiOR91 from a Culex quinquefasciatus larval head RNA sample, we found 2 alleles of this OR, differing at 9 residues. Functional analysis using the Xenopus oocyte expression system and 2-electrode voltage clamp electrophysiology revealed one allele (CquiOR91.1) to be nonfunctional, whereas the other allele (CquiOR91.2) was functional. Receptors formed by CquiOR91.2 and Cqui\Orco responded to (-)-fenchone, (+)-fenchone, and DEET, similar to what has been reported for Agam\Or40. We also identified 5 novel odorant ligands for the CquiOR91.2 + Cqui\Orco receptor: 2-isobutylthiazole, veratrole, eucalyptol, d-camphor, and safranal, with safranal being the most potent. To explore possible reasons for the lack of function for CquiOR91.1, we generated a series of mutant CquiOR91.2 subunits, in which the residue at each of the 9 polymorphic residue positions was changed from what occurs in CquiOR91.2 to what occurs in CquiOR91.1. Eight of the 9 mutant versions of CquiOR91.2 formed functional receptors, responding to (-)-fenchone. Only the CquiOR91.2 Y183C mutant was nonfunctional. The reverse mutation (C183Y) conferred function on CquiOR91.1 , which became responsive to (-)-fenchone and safranal. These results indicate that the "defect" in CquiOR91.1 that prevents function is the cysteine at position 183.


Subject(s)
Culicidae/chemistry , Insect Proteins/genetics , Receptors, Odorant/genetics , Alleles , Animals , Camphanes , DEET/metabolism , Insect Proteins/metabolism , Ligands , Mutation , Norbornanes/metabolism , Protein Subunits , Receptors, Odorant/metabolism
10.
Cell Mol Life Sci ; 74(2): 319-338, 2017 01.
Article in English | MEDLINE | ID: mdl-27535661

ABSTRACT

Anopheles gambiae Odorant Binding Protein 1 in complex with the most widely used insect repellent DEET, was the first reported crystal structure of an olfactory macromolecule with a repellent, and paved the way for OBP1-structure-based approaches for discovery of new host-seeking disruptors. In this work, we performed STD-NMR experiments to directly monitor and verify the formation of a complex between AgamOBP1 and Icaridin, an efficient DEET alternative. Furthermore, Isothermal Titration Calorimetry experiments provided evidence for two Icaridin-binding sites with different affinities (Kd = 0.034 and 0.714 mM) and thermodynamic profiles of ligand binding. To elucidate the binding mode of Icaridin, the crystal structure of AgamOBP1•Icaridin complex was determined at 1.75 Å resolution. We found that Icaridin binds to the DEET-binding site in two distinct orientations and also to a novel binding site located at the C-terminal region. Importantly, only the most active 1R,2S-isomer of Icaridin's equimolar diastereoisomeric mixture binds to the AgamOBP1 crystal, providing structural evidence for the possible contribution of OBP1 to the stereoselectivity of Icaridin perception in mosquitoes. Structural analysis revealed two ensembles of conformations differing mainly in spatial arrangement of their sec-butyl moieties. Moreover, structural comparison with DEET indicates a common recognition mechanism for these structurally related repellents. Ligand interactions with both sites and binding modes were further confirmed by 2D 1H-15N HSQC NMR spectroscopy. The identification of a novel repellent-binding site in AgamOBP1 and the observed structural conservation and stereoselectivity of its DEET/Icaridin-binding sites open new perspectives for the OBP1-structure-based discovery of next-generation insect repellents.


Subject(s)
Anopheles/metabolism , Insect Repellents/chemistry , Insect Repellents/metabolism , Piperidines/chemistry , Receptors, Odorant/chemistry , Receptors, Odorant/metabolism , Animals , Calorimetry , Crystallography, X-Ray , DEET/chemistry , DEET/metabolism , Fluorescence , Hydrogen Bonding , Models, Molecular , Piperidines/metabolism , Protein Binding , Protein Multimerization , Proton Magnetic Resonance Spectroscopy , Solutions , Static Electricity , Stereoisomerism
12.
Curr Biol ; 24(15): 1723-30, 2014 Aug 04.
Article in English | MEDLINE | ID: mdl-25042590

ABSTRACT

Dopaminergic neurons provide value signals in mammals and insects. During Drosophila olfactory learning, distinct subsets of dopaminergic neurons appear to assign either positive or negative value to odor representations in mushroom body neurons. However, it is not known how flies evaluate substances that have mixed valence. Here we show that flies form short-lived aversive olfactory memories when trained with odors and sugars that are contaminated with the common insect repellent DEET. This DEET-aversive learning required the MB-MP1 dopaminergic neurons that are also required for shock learning. Moreover, differential conditioning with DEET versus shock suggests that formation of these distinct aversive olfactory memories relies on a common negatively reinforcing dopaminergic mechanism. Surprisingly, as time passed after training, the behavior of DEET-sugar-trained flies reversed from conditioned odor avoidance into odor approach. In addition, flies that were compromised for reward learning exhibited a more robust and longer-lived aversive-DEET memory. These data demonstrate that flies independently process the DEET and sugar components to form parallel aversive and appetitive olfactory memories, with distinct kinetics, that compete to guide learned behavior.


Subject(s)
Conditioning, Classical , Drosophila melanogaster/physiology , Learning , Odorants , Animals , Appetitive Behavior , Avoidance Learning , Carbohydrates/physiology , DEET/metabolism , Female , Male , Olfactory Perception
13.
Nature ; 502(7472): 507-12, 2013 Oct 24.
Article in English | MEDLINE | ID: mdl-24089210

ABSTRACT

There are major impediments to finding improved DEET alternatives because the receptors causing olfactory repellency are unknown, and new chemicals require exorbitant costs to determine safety for human use. Here we identify DEET-sensitive neurons in a pit-like structure in the Drosophila melanogaster antenna called the sacculus. They express a highly conserved receptor, Ir40a, and flies in which these neurons are silenced or Ir40a is knocked down lose avoidance to DEET. We used a computational structure-activity screen of >400,000 compounds that identified >100 natural compounds as candidate repellents. We tested several and found that most activate Ir40a(+) neurons and are repellents for Drosophila. These compounds are also strong repellents for mosquitoes. The candidates contain chemicals that do not dissolve plastic, are affordable and smell mildly like grapes, with three considered safe in human foods. Our findings pave the way to discover new generations of repellents that will help fight deadly insect-borne diseases worldwide.


Subject(s)
DEET/metabolism , Insect Repellents/metabolism , Receptors, Odorant/metabolism , Sensory Receptor Cells/metabolism , Animals , Arthropod Antennae/anatomy & histology , Arthropod Antennae/cytology , Arthropod Antennae/drug effects , Arthropod Antennae/metabolism , Avoidance Learning/drug effects , Computer Simulation , Culicidae/drug effects , Culicidae/physiology , DEET/pharmacology , Drosophila melanogaster/cytology , Drosophila melanogaster/drug effects , Drosophila melanogaster/metabolism , Drosophila melanogaster/physiology , Humans , Insect Repellents/adverse effects , Insect Repellents/pharmacology , Sensory Receptor Cells/drug effects
14.
Anal Chim Acta ; 787: 267-73, 2013 Jul 17.
Article in English | MEDLINE | ID: mdl-23830449

ABSTRACT

Human exposure to N,N-diethyl-m-toluamide (DEET) occurs because of the widespread use of DEET as an active ingredient in insect repellents. However, information on the extent of such exposure is rather limited. Therefore, we developed a fast on-line solid phase extraction-high performance liquid chromatography-isotope dilution tandem mass spectrometry (HPLC-MS/MS) method to measure in urine the concentrations of DEET and two of its oxidative metabolites: N,N-diethyl-3-(hydroxymethyl)benzamide and 3-(diethylcarbamoyl)benzoic acid (DCBA). To the best of our knowledge, this is the first HPLC-MS/MS method for the simultaneous quantification of DEET and its select metabolites in human urine. After enzymatic hydrolysis of the conjugated species in 0.1 mL of urine, the target analytes were retained and pre-concentrated on a monolithic column, separated from each other and from other urinary biomolecules on a reversed-phase analytical column, and detected by atmospheric pressure chemical ionization in positive ion mode. The limits of detection ranged from 0.1 ng mL(-1) to 1.0 ng mL(-1), depending on the analyte. Accuracy ranged between 90.4 and 104.9%, and precision ranged between 5.5 and 13.1% RSD, depending on the analyte and the concentration. We tested the usefulness of this method by analyzing 75 urine samples collected anonymously in the Southeastern United States in June 2012 from adults with no known exposure to DEET. Thirty eight samples (51%) tested positive for at least one of the analytes. We detected DCBA most frequently and at the highest concentrations. Our results suggest that this method can be used for the analysis of a large number of samples for epidemiological studies to assess human exposure to DEET.


Subject(s)
DEET/urine , Oxidative Stress/physiology , Solid Phase Extraction/methods , Tandem Mass Spectrometry/methods , Chromatography, High Pressure Liquid/methods , DEET/metabolism , Female , Humans , Male
15.
Int J Pharm ; 454(2): 633-40, 2013 Oct 01.
Article in English | MEDLINE | ID: mdl-23732393

ABSTRACT

N,N-diethyl-meta-toluamide (DEET) is a widely used insect repellent due to its high efficacy. In this work, micellar systems based on poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer were developed and studied for the purpose of controlling the release and cutaneous permeation of DEET, using concentrated solutions of the copolymer Pluronic F127 to form thermoreversible gels. The formulations presented thermoreversible gelation above 5°C and altered rheological behavior at 15 and 25°C. The presence of the drug drastically changed the sol-gel transition temperatures. The micrographs suggest that DEET induced the formation of anisotropic structures, and Maltese Crosses were observed. The formulation containing 10wt% DEET and 15wt% Pluronic F127 presented sustained drug release for up to 7h. DEET release profile followed the Higuchi kinetics model. There was a reduction of approximately 35% in the amount of DEET absorbed through the skin after 6h. About 62% of DEET from the formulation consisting of Pluronic F127 and DEET remain retained on the skin. The anisotropic structure may constitute a barrier to diffusion and thereby controlling the drug release effectively. These tests suggest that the tested samples exhibit safety profile greater than some commercially available products.


Subject(s)
DEET/chemistry , Insect Repellents/chemistry , Micelles , Polyethylene Glycols/chemistry , Propylene Glycols/chemistry , Animals , DEET/metabolism , Delayed-Action Preparations/chemistry , In Vitro Techniques , Insect Repellents/metabolism , Mice , Poloxamer/chemistry , Polyethylene Glycols/metabolism , Propylene Glycols/metabolism , Rheology , Skin/metabolism , Skin Absorption , Solubility
16.
J Biol Chem ; 288(6): 4475-85, 2013 Feb 08.
Article in English | MEDLINE | ID: mdl-23261834

ABSTRACT

The Anopheles gambiae mosquito, which is the vector for Plasmodium falciparum malaria, uses a series of olfactory cues emanating from human sweat to select humans as their source for a blood meal. Perception of these odors within the mosquito olfactory system involves the interplay of odorant-binding proteins (OBPs) and odorant receptors and disrupting the normal responses to those odorants that guide mosquito-human interactions represents an attractive approach to prevent the transmission of malaria. Previously, it has been shown that DEET targets multiple components of the olfactory system, including OBPs and odorant receptors. Here, we present the crystal structure of A. gambiae OBP1 (OBP1) in the complex it forms with a natural repellent 6-methyl-5-heptene-2-one (6-MH). We find that 6-MH binds to OBP1 at exactly the same site as DEET. However, key interactions with a highly conserved water molecule that are proposed to be important for DEET binding are not involved in binding of 6-MH. We show that 6-MH and DEET can compete for the binding of attractive odorants and in doing so disrupt the interaction that OBP1 makes with OBP4. We further show that 6-MH and DEET can bind simultaneously to OBPs with other ligands. These results suggest that the successful discovery of novel reagents targeting OBP function requires knowledge about the specific mechanism of binding to the OBP rather than their binding affinity.


Subject(s)
Anopheles/chemistry , DEET/chemistry , Insect Proteins/chemistry , Insect Repellents/chemistry , Receptors, Odorant/chemistry , Animals , Anopheles/genetics , Anopheles/metabolism , Crystallography, X-Ray , DEET/metabolism , Humans , Insect Proteins/genetics , Insect Proteins/metabolism , Insect Repellents/metabolism , Insect Vectors/chemistry , Insect Vectors/genetics , Insect Vectors/metabolism , Ketones/chemistry , Ketones/metabolism , Plasmodium falciparum , Protein Binding , Protein Structure, Tertiary , Receptors, Odorant/genetics , Receptors, Odorant/metabolism
17.
J Control Release ; 160(3): 502-8, 2012 Jun 28.
Article in English | MEDLINE | ID: mdl-22546679

ABSTRACT

N,N-diethyl-3-methylbenzamide (DEET) is popular insect repellent which is considered safe and effective, yet is subject to considerable skin absorption. Skin absorption decreases effective repellency since less DEET is available for evaporation. We have investigated the extent to which DEET skin absorption can be reduced and evaporation sustained through encapsulation. DEET permeation through human skin in vitro was measured for an ethanolic solution standard and for two novel topical controlled-release formulations in which the DEET active material was temporarily sequestered within a permeable, charged-film microcapsule. Evaporation measurements were gathered using Tenax TA cartridges and a sampling pump drawing air over the skin. Three formulations were studied: a previously reported microcapsule formulation (Formulation A); a newly-developed microcapsule formulation (Formulation B); and a non-encapsulated ethanol control solution. Formulation B led to a 30% reduction in DEET permeation versus control. The two microcapsule DEET formulations exhibited 36-40% higher cumulative evaporation from the skin than did the control. The vapor trapping measurements in vitro show that Formulation B provided more than 48h of effective evaporation rate for repellency, while Formulation A provided less than 35h and the ethanol control less than 15h. This establishes a technical advantage for the controlled-release approach.


Subject(s)
DEET/chemistry , DEET/metabolism , Insect Repellents/chemistry , Insect Repellents/metabolism , DEET/administration & dosage , Ethanol/administration & dosage , Ethanol/chemistry , Ethanol/metabolism , Humans , In Vitro Techniques , Insect Repellents/administration & dosage , Skin Absorption , Volatilization
18.
FEBS J ; 279(6): 1044-53, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22251573

ABSTRACT

The initial step in the biodegradation pathway of N,N-diethyl-m-toluamide (DEET) in Pseudomonas putida strain DTB is catalyzed by DEET hydrolase (DthA), which hydrolyzes the amide bond to yield 3-methylbenzoic acid and diethylamine. In order to extend our understanding of DthA, the enzyme was purified and characterized. The enzyme is most active at pH 7.9, and is probably a tetramer in its native state. The kinetic parameters of the wild-type enzyme are K(m) = 10.2 ± 0.8 µm, k(cat) = 5.53 ± 0.09 s(-1) , and k(cat) /K(m) = (5.4 ± 0.4) × 10(5) m(-1) ·s(-1) . Mild substrate inhibition was observed with DEET concentrations over 500 µm. A homology model of DthA was used to guide mutational analysis of the active site, confirming that the catalytic triad is formed by Ser166, Ap292, and His320. The oxyanion hole is formed by the side chain OH of Tyr84 and the backbone amide of Trp167, with the Tyr84 OH being essential for enzyme activity. The DthA model also revealed a hydrophobic substrate-binding pocket comprosed of Trp167, Met170, and Trp214. W167A and M170A mutations decreased enzymatic activity and exacerbated substrate inhibition, whereas Trp214, which probably plays a role in substrate recognition, was essential for enzymatic activity. The pH rate profile of DthA was fitted to two ionizable groups (pK(a1) = 6.1 and pK(a2) = 9.9) that probably correspond to Nε of His320 and the OH of Tyr84, respectively. In addition to catalyzing the hydrolysis of DEET, DthA hydrolyzed a variety of esters and amides.


Subject(s)
Bacterial Proteins/chemistry , Hydrolases/chemistry , Pseudomonas putida/enzymology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Catalysis , Catalytic Domain , DEET/metabolism , Glutamine/genetics , Glutamine/metabolism , Hydrogen-Ion Concentration , Hydrolases/genetics , Hydrolases/metabolism , Hydrolysis , Kinetics , Mutagenesis, Site-Directed , Substrate Specificity
19.
Integr Environ Assess Manag ; 8(1): 120-34, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22006575

ABSTRACT

N,N-diethyl-m-toluamide (DEET) is a key active ingredient in many insect repellents available commercially throughout the world. Owing to its popularity among consumers for nearly 30 years, considerable work conducted in the past has demonstrated-and continues to demonstrate-that human exposure to DEET poses no significant health risk to the general population. The results of several studies reported in this paper describe more recent work to understand the environmental fate of DEET, particularly in surface waters and soil, and the potential hazards to aquatic and terrestrial organisms. In summary, DEET enters the environment through several pathways: directly into air during spray application; to surface water from overspray and indirectly via wastewater treatment plant (WTTP) discharges (as a result of washing of skin and laundering of clothing); or to soil via overspray and application of treated sewage as an amendment. Multimedia environmental fate modeling predicts that DEET entering the environment is retained either in receiving waters (∼79%) or in soil (∼21%). Based on its physicochemical properties, DEET is expected to be moderately mobile in the soil column. In surface waters and soil, DEET degrades at a moderate to rapid rate (its half-life is measured in days to weeks). The small amounts of DEET retained in air are subject to rapid photo-oxidation via hydroxyl radical-mediated degradation or, if in droplet form, gravitational settling to soil or water. DEET does not interfere with ozone formation in the upper atmosphere. The bioaccumulation potential of DEET is low; it is neither a persistent, bioaccumulative toxicant nor a persistent organic pollutant. Among aquatic species, acute effect concentrations range between 4 and 388 mg/L. The chronic no-observed effect concentrations (NOEC) for daphnids and green algae range from approximately 0.5 to 24 mg/L. Measured concentrations of DEET in surface waters are several hundreds to thousands of times lower than the lowest NOEC measured, and thus the probability for adverse effects to environmental species is low. A separate paper by Aronson et al. (this issue) supports this conclusion by quantitatively exploring the risks to the aquatic environment using a combination of monitoring data and exposure modeling.


Subject(s)
DEET/metabolism , DEET/toxicity , Environmental Pollutants/metabolism , Environmental Pollutants/toxicity , Insect Repellents/metabolism , Insect Repellents/toxicity , Animals , Aquatic Organisms/drug effects , Aquatic Organisms/metabolism , Colinus/metabolism , DEET/chemistry , Environmental Monitoring/methods , Environmental Pollutants/chemistry , Insect Repellents/chemistry , Models, Biological , Species Specificity
20.
Chemosphere ; 80(7): 687-92, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20580410

ABSTRACT

Constructed treatment wetlands have the potential to reclaim wastewaters through removal of trace concentrations of emerging organic pollutants, including pharmaceuticals, personal care products, and pesticides. Flask-scale assessments incorporating active and inactivated duckweed were used to screen for plant-associated removal of emerging organic pollutants in aquatic plant systems. Removals of four of eight pollutants, specifically atrazine, meta-N,N-diethyl toluamide (DEET), picloram, and clofibric acid, were negligible in all experimental systems, while duckweed actively increased aqueous depletion of fluoxetine, ibuprofen, 2,4-dichlorophenoxyacetic acid, and triclosan. Active plant processes affecting depletion of experimental pollutants included enhancement of microbial degradation of ibuprofen, uptake of fluoxetine, and uptake of degradation products of triclosan and 2,4-dichlorophenoxyacetic acid. Passive plant processes, particularly sorption, also contributed to aqueous depletion of fluoxetine and triclosan. Overall, studies demonstrated that aquatic plants contribute directly and indirectly to the aqueous depletion of emerging organic pollutants in wetland systems through both active and passive processes.


Subject(s)
Araceae/metabolism , Cosmetics/metabolism , Pesticides/metabolism , Pharmaceutical Preparations/metabolism , Water Pollutants, Chemical/metabolism , 2,4-Dichlorophenoxyacetic Acid/metabolism , Atrazine/metabolism , Biodegradation, Environmental , Clofibric Acid/metabolism , DEET/metabolism , Fluoxetine/metabolism , Ibuprofen/metabolism , Picloram/metabolism , Triclosan/metabolism , Wetlands
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