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PubMed1988

Inhibition of metabolic cooperation in vitro and enhancement of enzyme altered foci incidence in rat liver by the pyrethroid insecticide fenvalerate.

The synthetic pyrethroids cypermethrin, delta-methrin, fenvalerate, permethrin, and the fenvalerate metabolite p-chlorophenylisovaleric acid were investigated for inhibition of gap-junctional intercellular communication in vitro in the Chinese hamster lung fibroblast (V79) metabolic cooperation assay. Fenvalerate was furthermore studied for enhancement of gamma-glutamyl transpeptidase-positive enzyme altered foci incidence in partially hepatectomized, nitrosodiethylamine-initiated male Sprague Dawley rats. The in vitro studies showed that fenvalerate and p-chlorophenylisovaleric acid were inhibitors of intercellular communication at non-cytotoxic concentrations while cypermethrin, deltamethrin, and permethrin were inactive. In the in vivo study in rat liver, fenvalerate administered p.o. (75 mg/kg/day) 5 days a week for 10 weeks induced significantly more foci per cm3 and a larger percentage of liver tissue occupied by foci tissue compared to a vehicle control group. Analysis of size distributions of foci in fenvalerate- and vehicle-treated rats showed elevated foci incidences in fenvalerate-treated rats at all foci sizes. Fenvalerate induced no hepatotoxic effects as judged by plasma transaminase activities and histopathology. The results of this study suggest fenvalerate to be a potential tumour promoter.

Animals↗
PubMed2006

Monitoring of nutrients, pesticides, and metals in waters, sediments, and fish of a wetland.

Wetland areas are of extraordinary importance for the conservation of wildlife. The Aiguamolls de l'Empordà Natural Park, located in Girona (northeast Spain), is one of the few areas in Europe acting as a way station for migratory birds. The natural park is made up of a brackish water reserve and a fresh water reserve. Agriculture and tourism, which are concentrated especially around coastal population centers, are the main activities in this area and result in the release into the environment of nutrients, pesticides, and heavy metals. This article aims to investigate the presence of nutrients, selected pesticides (organochlorine compounds, permethrin and triazines) and metals (Cr, Cu, Cd, Ni and Pb) in water, sediments, and fish samples. In the case of water, seasonal variations in levels of contamination were also monitored. Comparison was made of the fresh and brackish water reserves and concentration factors for metals and pesticides in sediment were determined. We conclude that the most significant sources of contamination in the natural park are from the entry of pesticides and nutrients into surface waters and sediments as a result of the intensive farming activity of the surrounding areas. The pesticides with the greatest presence were found to be lindane, heptachlor epoxide, permethrin, and atrazine. Among the metals analyzed, Cu and Cr presented the highest concentrations in surface waters and sediments.

Animals↗
PubMed2006

Essential oils as potential adulticides against two populations of Aedes aegypti, the laboratory and natural field strains, in Chiang Mai province, northern Thailand.

Essential oils derived from five plant species, celery (Apium graveolens), caraway (Carum carvi), zedoary (Curcuma zedoaria), long pepper (Piper longum), and Chinese star anise (Illicium verum), were subjected to investigation of adulticidal activity against mosquito vectors. Two populations of Aedes aegypti, the laboratory and natural field strains, collected in Chiang Mai province, northern Thailand were tested in pyrethroid-susceptibility bioassays. The results revealed that the natural field strain of A. aegypti was resistant to permethrin, with mortality rates ranging from 51 to 66%. A mild susceptibility, with mortality rates ranging from 82 to 88%, was observed in the natural field strain of A. aegypti exposed to lambdacyhalothrin, which suggested that this strain was tolerant and might be resistant to this insecticide. However, laboratory-reared A. aegypti exposed to discriminating dosages of permethrin and lambdacyhalothrin induced 100% mortality in all cases, thus indicating complete susceptibility of this strain to these insecticides. The adulticidal activity determined by topical application revealed that all five essential oils exerted a promising adulticidal efficacy against both laboratory and natural field strains of A. aegypti. Although the laboratory strain was slightly more susceptible to these essential oils than the natural field strain, no statistically significant difference was observed. Moreover, comparison of the adulticidal activity indicated that the performance of these essential oils against the two strains of A. aegypti was similar. The highest potential was established from caraway, followed by zedoary, celery, long pepper, and Chinese star anise, with an LC(50) in the laboratory strain of 5.44, 5.94, 5.96, 6.21, and 8.52 microg/mg female, respectively, and 5.54, 6.02, 6.14, 6.35, and 8.83 microg/mg female, respectively, in the field strain. These promising essential oils are, therefore, an alternative in developing and producing mosquito adulticides as an effective measure used in controlling and eradicating mosquito vectors.

Aedes↗
PubMed1995

The suitability of the micronucleus assay in human lymphocytes as a new biomarker of excision repair.

The cytokinesis blocked micronucleus assay is relatively insensitive to detect agents that predominantly induce excision repairable DNA lesions. However, it has been recently proposed that excision-repairable DNA lesions induced in G0/G1 phase can be converted to micronuclei by using inhibitors of the gap filling step of excision repair so that unfilled gaps are converted to double stranded breaks after S phase and micronuclei (MN) at completion of mitosis. As it has been recently demonstrated this process could be improved by combining cytosine arabinoside (ARA-C) and hydroxyurea (HU). In the present work, we have investigated the suitability of this new approach by studying its ability to detect excision repairable DNA lesions induced by 10 pesticides (alachlor, atrazine, cypermethrin, deltamethrin, fenpropathrin, fenvalerate, maleic hydrazide, paraquat, permethrin and trifluralin) and 3 well-known mutagenic agents (ethyl methane sulphonate, EMS; methylnitrosourea, MNU; and mytomicin C, MMC). Our results showed that the combination of ARA-C and HU substantially increased the level of MN in whole blood lymphocyte cultures, but it provided an excess of toxicity when further treatments, such as MNU, were performed. When ARA-C alone was used, the ARA/CBMN assay appeared to be highly sensitive and specific in detecting agents known to induce excision repairable DNA lesions. Thus, EMS and MNU but not MMC greatly induced DNA excision repair. On the other hand, alachlor, permethrin and, to a lesser extent, trifluralin and fenpropathrin also increased the ratio of excision repairable DNA lesions converted to MN. On the contrary, atrazine, cypermethrin, deltamethrin, fenvalerate, maleic hydrazide and paraquat did not induce excision repair.

Adult↗
PubMed1988

The effects of type I and II pyrethroids on motor activity and the acoustic startle response in the rat.

Recent data have demonstrated that the in vivo effects of low dosages of two pyrethroids, cismethrin and deltamethrin, can be differentiated. Two behavioral tests, locomotor activity and the acoustic startle response (ASR), were utilized to separate the behavioral actions of Type I and II pyrethroids using permethrin, RU11679, cypermethrin, RU26607, fenvalerate, cyfluthrin, flucythrinate, fluvalinate and p,p'-DDT. Dosage-effect functions for all compounds were determined for both figure-eight-maze activity and the ASR in the rat. All compounds were administered po in 1 ml/kg corn oil 1.5-3 hr prior to testing. All compounds produced dosage-dependent decreases in locomotor activity. The Type I compounds, permethrin and RU11679, along with p,p'-DDT, increased amplitude and had no effect on latency to onset of the ASR. In contrast, the Type II pyrethroids, cypermethrin, cyfluthrin, and flucythrinate, decreased amplitude and increased the latency to onset of the ASR. Fenvalerate increased the amplitude, had no effect on latency, but unlike the other compounds tested, increased ASR sensitization. Fluvalinate had no effect on any measure of the ASR. These data provide further evidence of the differences between the in vivo effects of low dosages of Type I and II pyrethroids, and extend the findings of our previous work to other representatives of the two classes of pyrethroids.

Acoustic Stimulation↗
PubMed2005

Bionomics of Anopheles stephensi Liston in the malarious area of Hormozgan province, southern Iran, 2002.

Anopheles stephensi Liston is an important malaria vector in Hormozgan province, where it is the most prevalent anopheline mosquito. It shows two annual activity peaks, one in spring and another in the autumn. In mountainous areas the second peak starts earlier than in coastal regions. Adults are endophilic and endophagic, but in the hot season when people sleep outside buildings they frequently bite outdoors. Larvae are found in a wide-range of habitats, both natural and man-made. All three biological forms of the species, occur in the province, i.e. An. stephensi stephensi (type form), An. stephensi mysorensis, and the intermediate form. An. stephensi mysorensis is found only in rural-mountainous areas, whereas the type and intermediate forms occur in urban-coastal regions and the rural plains, with the type form predominant. The presence of the type form in urban areas and mysorensis in rural areas is consistent with the available epidemiological data for malaria in the region and with the finding in India that the type form is an efficient malaria vector inhabiting urban areas whereas mysorensis is rural and has a lower vectorial capacity. Insecticide susceptibility tests on field collected adult mysorensis and adults from laboratory strains of the type and intermediate forms were carried out according to WHO standard methods. These showed that all three forms are susceptible to bendiocarb, propoxur, malathion, fenitrothion, deltamethrin, permethrin, cyfluthrin, and lambdacyhalothrin, but are resistant to DDT and show low level of tolerance to dieldrin. Examination of the larvicidal activity of malathion, fenitrothion, temephos and chlorpyrifos at diagnostic doses showed that these stephensi forms are susceptible to all larvicides except fenitrothion. Irritability tests to pyrethroid insecticides showed high levels of irritability to permethrin and lambdacyhalothrin, but low irritability to cyfluthrin and deltamethrin. The importance of these findings for the epidemiology and control of malaria in the region are discussed.

Animals↗
PubMed2006

Pilot survey monitoring pharmaceuticals and related compounds in a sewage treatment plant located on the Mediterranean coast.

A one-year monitoring study was performed to evaluate the occurrence, persistence and fate of a group of 14 organic compounds in a sewage treatment plant (STP) located in the south of Spain. These results are part of a more extensive study, financed by the Spanish Ministry of Research with the aim to evaluate the traceability of new pollutants on the Mediterranean coast and to determine the removal efficiency of sewage treatment plants (STP) for these pollutants. The compounds which have been analyzed include pharmaceuticals of various therapeutic categories (ibuprofen, acetaminophen, dipyrone, diclofenac, carbamazepine and codeine), pesticides (chlorfenvinfos and permethrin), caffeine, triclosan, bisphenol A and three of their more relevant metabolites (1,7-dimethylxanthine, carbamazepine 10,11-epoxide and 2,7/2,8-dichlorodibenzo-p-dioxin). An SPE/GC-MS multi-residue analytical method was developed and validated to facilitate simultaneous determination of these compounds in both influent and effluent wastewater. The method provided mean recoveries higher than 75%, with the exception of 2,7/2,8-dichlorodibenzo-p-dioxin, dipyrone and permethrin which exhibited recoveries lower than 22%. The overall variability of the method was below 14%. The method detection limit (LOD) was between 1 and 100 ng l(-1) and precision, which was calculated as relative standard deviation (RSD), ranged from 1.8% to 11.2%. The application of the proposed method has allowed the identification of all the target compounds at mean concentrations which ranged from 0.12 to 134 microg l(-1) in the influent and from 0.09 to 18.0 microg l(-1) in the effluent. The removal efficiencies of the STP for these compounds varied from 20% (carbamazepine) to 99% (acetaminophen), but in all cases resulted insufficient in order to avoid their presence in treated water and subsequently in the environment.

Environmental Monitoring↗
PubMed2005

Identifying primary stressors impacting macroinvertebrates in the Salinas River (California, USA): relative effects of pesticides and suspended particles.

Laboratory dose-response experiments with organophosphate and pyrethroid pesticides, and dose-response experiments with increasing particle loads were used to determine which of these stressors were likely responsible for the toxicity and macroinvertebrate impacts previously observed in the Salinas River. Experiments were conducted with the amphipod Hyalella azteca, the baetid mayfly Procloeon sp., and the midge Chironomus dilutus (Shobanov, formerly Chironomus tentans). The results indicate the primary stressor impacting H. azteca was pesticides, including chlorpyrifos and permethrin. The mayfly Procloeon sp. was sensitive to chlorpyrifos and permethrin within the range of concentrations of these pesticides measured in the river. Chironomus dilutus were sensitive to chlorpyrifos within the ranges of concentrations measured in the river. None of the species tested were affected by turbidity as high as 1000 NTUs. The current study shows that pesticides are more important acute stressors of macroinvertebrates than suspended sediments in the Salinas River.

Amphipoda↗
PubMed2005

Structure-activity relationships for the action of 11 pyrethroid insecticides on rat Na v 1.8 sodium channels expressed in Xenopus oocytes.

Pyrethroid insecticides bind to voltage-sensitive sodium channels and modify their gating kinetics, thereby disrupting nerve function. This paper describes the action of 11 structurally diverse commercial pyrethroid insecticides on the rat Na v 1.8 sodium channel isoform, the principal carrier of the tetrodotoxin-resistant, pyrethroid-sensitive sodium current of sensory neurons, expressed in Xenopus laevis oocytes. All 11 compounds produced characteristic sodium tail currents following a depolarizing pulse that ranged from rapidly-decaying monoexponential currents (allethrin, cismethrin and permethrin) to persistent biexponential currents (cyfluthrin, cyhalothrin, cypermethrin and deltamethrin). Tail currents for the remaining compounds (bifenthrin, fenpropathrin, fenvalerate and tefluthrin) were monoexponential and decayed with kinetics intermediate between these extremes. Reconstruction of currents carried solely by the pyrethroid-modified subpopulation of channels revealed two types of pyrethroid-modified currents. The first type, found with cismethrin, allethrin, permethrin and tefluthrin, activated relatively rapidly and inactivated partially during a 40-ms depolarization. The second type, found with cypermethrin, cyfluthrin, cyhalothrin, deltamethrin, fenpropathrin and fenvalerate, activated more slowly and did not detectably inactivate during a 40-ms depolarization. Only bifenthrin did not produce modified currents that fit clearly into either of these categories. In all cases, the rate of activation of modified channels was strongly correlated with the rate of tail current decay following repolarization. Modification of Na v 1.8 sodium channels by cyfluthrin, cyhalothrin, cypermethrin and deltamethrin was enhanced 2.3- to 3.4-fold by repetitive stimulation; this effect appeared to result from the accumulation of persistently open channels rather than preferential binding to open channel states. Fenpropathrin was the most effective compound against Na v 1.8 sodium channels from the perspective of either resting or use-dependent modification. When use dependence is taken into account, cypermethrin, deltamethrin and tefluthrin approached the effectiveness of fenpropathrin. The selective expression of Na v 1.8 sodium channels in nociceptive neurons suggests that these channels may be important targets for pyrethroids in the production of paresthesia following dermal exposure.

Animals↗
PubMed1985

Interactions of pyrethroids with phosphatidylcholine bilayers: comparisons in liposomal systems exhibiting large or small radii of curvature.

Pyrethroid interactions with dipalmitoyl phosphatidylcholine (DPPC) vesicles have been characterized in bilayers having large and small radii of curvature. The abilities of pyrethroids to alter the gel-fluid phase transition profiles were determined by steady state fluorescence anisotropy and phase-modulation lifetime techniques using the fluorescent probes cis- and trans-parinaric acid. Using the geometric isomers of parinaric acid as membrane probes, pyrethroids were found to lower the phase transition temperature (Tc) of DPPC large multilamellar vesicles with the same order of comparative effectiveness as previously reported using the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH). Permethrin had a greater depressive effect upon the Tc of DPPC in the small unilamellar vesicle (SUV) system than in the large multilamellar system. Conversely, allethrin was less effective in reducing the Tc of DPPC SUVs. The enhanced effect of permethrin in decreasing the Tc of DPPC SUVs was greatest in regions of more rigid lipid packing, as determined by trans-parinaric acid fluorescence parameters. The results indicate that changes in lipid packing configuration caused by differing bilayer radii of curvature may alter the interactive characteristics of pyrethroids with lipid membranes.

Diphenylhexatriene↗
PubMed2000

Molecular and biochemical survey of acaricide resistance mechanisms in larvae from Mexican strains of the southern cattle tick, Boophilus microplus.

We examined the larvae of several organophosphate and pyrethroid-resistant Mexican strains of Boophilus microplus using biochemical and molecular tests to investigate the mechanisms conferring acaricide resistance. The electrophoretic profiles of esterase activity in protein extracts from coumaphos and permethrin-resistant strains compared to the susceptible strain revealed distinct differences, which inhibitor studies attributed to carboxylesterases. Esterase hydrolysis assays showed significant enhancement of both total and permethrin hydrolysis in one pyrethroid-resistant strain, with no enhancement in two other strains with very high resistance to pyrethroids. Sequence analysis of sodium channel mRNA fragments in all pyrethroid-resistant strains determined that they did not possess the classic kdr and super-kdr mutations known to confer pyrethroid resistance in several insect species. Using reverse transcriptase-polymerase chain reaction (RT-PCR) with degenerate primers designed from conserved regions of insect esterase amino acid sequences, a B. microplus larval cDNA fragment was isolated whose deduced amino acid sequence was significantly similar to esterases from a wide range of species. In Northern blot RNA analysis the cDNA hybridized to a 2.1 kb mRNA that was abundant in all resistant strains except one, in which a very low abundance could provide a marker for the mechanism conferring resistance in this strain.

Journal Article↗
PubMed2002

Analysis and implications of aircraft disinsectants.

Aircraft disinsection is required by various countries. In-flight spraying with a 2% phenothrin aerosol exposes passengers and crew directly. Residual spaying uses a permethrin emulsions in the absence of passengers and crew and results in dermal and oral exposures. Exposed passengers and crew often complain of, skin rashes, respiratory problems, tingling and numbness in fingertips and lips and burning eyes. A number of formulations were analyzed for their constituents using GLC-Mass. spec. Volatile organic compounds (VOCs) were found in all aerosol preparations including, ethyl benzene and xylene isomers along with phenothrin. Residual sprays contained, cis-, and trans-, permethrins, palmidrol, and occasionally naphthalene. Headspace analysis found methylene chloride and hexene derivatives but not the active ingredients. The known synergistic effects between organophosphates and pyrethrins, based on carboxyesterases inhibition, can be expected in the presence of Tricresylphosphates (TCPs), constituents found in jet engine oils and in some hydraulic fluids. During oil seal failure, the presence of TCP in the ventilation air could explain the increased sensitivity of some crew members and passengers to disinsectants.

Aerosols↗
PubMed2004

Ectoparasites--the underestimated realm.

CONTEXT: Ectoparasitoses (infestations with parasites that live on or in the skin) can cause considerable morbidity. Whereas pediculosis and scabies are ubiquitous, cutaneous larva migrans and tungiasis (sand-flea disease) occur mainly in hot climates. The prevalence of ectoparasitoses in the general population is usually low, but can be high in vulnerable groups. Scientific knowledge on how to deal best with parasitic skin diseases in different settings is scanty, and evidence-based measures for control are not available. For head lice and scabies the situation is daunting, because resistance of Pediculus humanus capitis and Sarcoptes scabiei to insecticides is spreading and unpredictable. STARTING POINT: J Hunter and S Barker reported different patterns of resistance in schoolchildren in Brisbane, Australia: full resistance to malathion, permethrin, and pyrethrum in two schools, whereas head lice were susceptible to malathion and, to a lesser extent, to pyrethrums in three other schools (Parasitol Res 2003; 90: 476-78). K Yoon and colleagues found different resistance patterns in the USA and Ecuador (Arch Dermatol 2003; 139: 994-1000). Head lice from Florida were less susceptible to permethrin than those from Texas, and parasites from Ecuador were susceptible to both insecticides tested. WHERE NEXT? The occurrence of resistant pediculosis and scabies is expected to increase numerically and geographically. Clinicoepidemiological studies are urgently needed to identify the factors which govern the emergence and spread of strains of P humanus capitis and S scabiei that are resistant to insecticide or acaricide. Oral treatment with ivermectin could substitute for topically applied compounds, particularly in resource-poor communities where polyparasitism is common. A better understanding of local epidemiology is required to develop control measures. This knowledge has to be applied in combination with environmental sanitation, health education, and culturally acceptable interventions that are affordable by the underprivileged.

Drug Resistance↗
PubMed2002

Effects of pyrethroid insecticides and estrogen on WNT10B proto-oncogene expression.

Breast cancer is a serious illness affecting approximately one in nine women in the United States. Although an actual cause for breast cancer is unknown, genetic and environmental factors have been associated with its onset. Elevated levels of estrogen and heightened expression of the WNT10B proto-oncogene have been implicated in the development of human malignant breast tumors because they enhance the proliferation of mammary tissue. Two pyrethroid insecticides, sumithrin and fenvalerate, have been shown to mimic estrogenic activity in MCF-7 human breast carcinoma cells by inducing pS2 expression whereas two other pyrethroids, permethrin and d-trans allethrin do not have the same capability. To investigate if estrogen and these four pyrethroid insecticides could affect the expression of a gene related to mammary gland development, WNT10B expression in pyrethroid-treated MCF-7 cells was examined. MCF-7 cells under normal growth conditions do not express WNT10B. Reverse-transcriptase polymerase chain reaction (RT-PCR), nested PCR and Southern hybridization were employed to detect WNT10B expression. As controls, cells were treated with either ethanol, corn oil, or Vista LPA solvent. When compared to the solvent-treated controls, sumithrin, fenvalerate and estrogen treated MCF-7 cells all had increased levels of WNT10B expression. The non-estrogenic pyrethroids, d-trans allethrin and permethrin, demonstrated a similar elevation of WNT10B expression at a lower concentration, but not at the higher concentration. The results suggest that pyrethroid insecticides and estrogen can enhance the expression of the WNT10B proto-oncogene. However, since both the estrogenic and non-estrogenic substances amplified Wnt10B expression, the mechanism likely involves multiple distinct pathways.

Breast Neoplasms↗
PubMed1997

The Gambian National Impregnated Bednet Programme: evaluation of the 1994 cost recovery trial.

Following the success of a controlled trial of insecticide-impregnated bednets in reducing mortality in children. The Gambia started a National Impregnated Bednet Programme (NIBP) in 1992. The objectives of this programme were to introduce impregnated bednets into all primary health care (PHC) villages and to establish a system of cost recovery over a three-year period. During the initial phase of the programme, when insecticide was given out free, a high uptake was achieved. However, after small user charges were introduced in 1993, coverage dropped to a low level. In 1994, different systems of insecticide distribution and permethrin formulations were tried in an attempt to improve coverage. A nationwide cross-sectional survey carried out during the 1994 rainy season measured coverage by distribution channel, as well as the knowledge, attitudes and practices of health workers and villagers during the intervention. Overall, only 16% of bednets were impregnated in 1994, compared to 80% when the insecticide was offered free of charge in previous years. Lack of money was the major reason given by villagers for not impregnating their bednets in 1994. Use of impregnated bednets was higher in areas where the sale of permethrin emulsion by village health workers was supplemented by the sale of insecticide in individual packages through shops. In villages where insecticide was distributed free to women with small children through governmental mother and child health (MCH) services, higher levels of coverage were achieved among women and young children than in villages where other distribution systems were used. We conclude that the sale of insecticide through the private sector may increase bednet impregnation rates in African communities, and that the free distribution of insecticide through MCH services may be an effective way of targeting young children, the group most at risk of malaria.

Adolescent↗
PubMed1999

Induction of mitotic cell division distrubances and mitotic arrest by pyrethroids in V79 cell cultures.

Five pyrethroids (fenvalerate, deltamethrin, cypermethrin, permethrin, cyfluthrin) differing in their chemical purity were investigated on their cytotoxic effects, especially on their ability to induce mitotic cell division disturbances using Chinese hamster lung cells of line V79. The colony forming ability (CFA) resulted in distinct differences of the cytotoxic effect of the tested pyrethroids, whereby permethrin was found to be most toxic. With the exception of fenvalerate all tested pyrethroids gave rise to inhibition of cell cycle progression as shown by G2/M-arrest of synchronized V79 cells by flow cytometry as well as by the increase of the mitotic index as evaluated by light microscopy. The mitotic arresting activity could be attributed to the occurrence of abnormal mitotic figures such as initial and full C-metaphases. The results however indicate, that pyrethroids per se do not contribute to the cytotoxic effects but that other factors such as chemical impurities, source as well as manufacturing process and isomer composition may be responsible for the observed cytotoxic effects.

Animals↗
PubMed2000

Molecular cloning, nucleotide sequence and gene expression of a cytochrome P450 (CYP6F1) from the pyrethroid-resistant mosquito, Culex quinquefasciatus Say.

To analyze cytochrome P450s in the southern house mosquito, Culex quinquefasciatus, we quantified the content of P450s and b5 in larval microsomes of guts and carcasses. Results indicated that content was 30 times higher in guts than in carcasses. A conserved region in the alignment of insect P450 family 6 (CYP6) proteins served as a guide for the synthesis of degenerate oligonucleotide primers to clone P450 cDNAs. Primers were used in the reverse transcription-polymerase chain reaction (RT-PCR) of gut mRNA from 4th-instar larvae of the permethrin-susceptible or resistant C. quinquefasciatus. PCR products of ca. 250 base pairs (bp) were cloned, and nucleotide sequences of 35 clones from susceptible and 28 from resistant strains determined. Alignment of the deduced amino acid sequences from these clones showed them to be classifiable into six isoforms. We next screened a cDNA clone (CYP6F1) from a gut cDNA library and determined the nucleotide sequence. Northern blot analysis showed that the CYP6PF1 gene in the permethrin-resistant strain appeared to be expressed more strongly than in the susceptible strain. The deduced amino acid of CYP6F1 showed that it has conserved domains of a membrane-anchoring signal, reductase binding sites, a heme-binding site, ETLR motif and substrate recognition sites in P450s. Phylogenetic analysis showed that CYP6F1 is strongly related to CYP6D1 involved in pyrethroid detoxification.

Amino Acid Sequence↗
PubMed2002

Human blood and environmental media screening method for pesticides and polychlorinated biphenyl compounds using liquid extraction and gas chromatography-mass spectrometry analysis.

Screening assessment methods have been developed for semi- and non-volatile persistent organic pollutants (POPs) for human blood and solid environmental media. The specific methodology is developed for measuring the presence of "native" compounds, specifically, a variety of organochlorine pesticides (OCPs), organophosphate pesticides (OPPs), and for polychlorinated biphenyls (PCBs). The method is demonstrated on anonymous Red Cross blood samples as well as two potential environmental sources, tracked in soil and dog hair. This work is based on previously developed methods for semi-volatile hydrocarbon exposure from fuels usage and similarly employs liquid solvent extraction, evaporative volume reduction. and subsequent specialized gas chromatography-mass spectrometry analysis (GC-MS). Standard curves, estimates of recovery efficiency, and specific GC-MS SIM quantification methods were developed for common pesticides including diazinon. aldrin, chlorpyrifos, malathion, dieldrin, DDT, permethrin, cyhalothrin, and cypermethrin, and for seven selected PCBs. Trace levels of certain PCBs and pesticides such as permethrin, dieldrin, malathion, lindane, diazinon, and chlorpyrifos were tentatively identified in anonymous blood samples as well as in two potential environmental sources. tracked in soil and dog hair. The method provides a simple screening procedure for various media and a variety of common organic pollutants without extensive sample preparation. It is meant to complement and augment data from more specific or complex methodology, to provide initial broad spectrum guidance for designing targeted experiments, and to provide confirmatory evidence for the usual metabolic biomarker measurements made to assess human exposure.

Environmental Monitoring↗