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J Hemingway

Publications and source records attributed to J Hemingway.

At least 73 records · Page 4Linked to original sources
PubMed1996

Purification and characterization of a major glutathione S-transferase from the mosquito Anopheles dirus (species B).

The major form of glutathione S-transferase (GST) activity from the mosquito Anopheles dirus (species B), a vector of malaria in Thailand has been purified 421-fold. It constituted approx. 20% of the total measured CDNB conjugating activity in the homogenate. This enzyme appeared as a single band of 25.0 +/- 0.26 kDa on SDS-PAGE and was kinetically characterized with 10 substrates and 4 inhibitors. The enzyme is capable of catalysing dehydrochlorination of 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (DDT) in vitro at a rate of 4.4 nmol of 1,1-dichloro-2,2-bis-(p-chlorophenyl)ethane (DDE) formation per mg protein. This is comparable to the rate of catalysis of the orthologous isoenzyme from An. gambiae reported previously. The IC50 plots of the inhibitor data (fractional velocity vs log [I]) for three of the inhibitors indicate the homogenous nature of this enzyme. However, inhibition by ethacrynic acid demonstrates more than a single affinity site for interaction. The six N-terminal amino acids of the purified enzyme are identical to a GST reported from Aedes aegypti, which was indicated to play a role in DDT-resistance in this species. The results suggest that the two enzymes may belong to the same class, however each possesses a different specificity.

Amino Acid Sequence↗
PubMed1996

Effect of fenthion treatment on larval densities of insecticide-resistant Culex quinquefasciatus in an urban area of Sri Lanka.

In Sri Lanka the national Anti-Filaria Campaign (AFC) has routinely employed fenthion since 1974 for larvicidal control of Culex quinquefasciatus, the vector of Bancroftian filariasis in urban areas, where this mosquito breeds prolifically in polluted waters. During 1994 the efficacy of AFC fenthion treatment against organophosphate-resistant Cx quinquefasciatus was investigated at Dehiwela, near Colombo. The AFC target rate of fenthion application was 1 mg a.i./l, but the actual concentrations of fenthion in freshly treated pits ranged from 0.64 to 63.2 mg a.i./l. There was significant suppression of larval densities in treated soakage pits, the predominant breeding site of Cx quinquefasciatus, although the mosquito population was 6-fold resistant to fenthion at the LC50 level. Production of pupae was almost completely prevented in soakage pits which were sprayed weekly with fenthion, indicating that adult mosquito emergence from this source was minimal. The rapid decline in concentration of fenthion detected in the water of soakage pits indicated that a weekly treatment schedule is essential for effective control. With the rapid recolonization of treated sites, the weekly schedule must be strictly implemented in order to achieve control of resistant larvae. Fenthion activity levels detected in treated pits suggest that a 7-10 day schedule of retreatment would completely suppress susceptible Cx quinquefasciatus.

Animals↗
PubMed1995

Kinetic and molecular differences in the amplified and non-amplified esterases from insecticide-resistant and susceptible Culex quinquefasciatus mosquitoes.

Two non-amplified esterases were purified from the insecticide-susceptible Pel SS strain of Culex quinquefasciatus. These were the two major esterase activity peaks in this strain. The two corresponding amplified carboxylesterases, Est alpha 2 and Est beta 2, involved in organophosphate sequestration were purified from two resistant C. quinquefasciatus strains. The Pel SS esterases were significantly less reactive with the organophosphates than those from the resistant strains. One of the Pel SS esterases was electrophoretically identical to amplified Culex Est beta 1. However, it differed kinetically, and in its nucleotide and predicted amino acid sequences from the two characterized amplified Est beta 1s, it is classified as Est beta 1(3). Restriction fragment analysis suggested Pel SS has only one Est alpha and one Est beta gene, while the resistant Pel RR has both amplified and non-amplified forms of Est alpha and Est beta. The EcoRI fragments for both Pel SS esterases were distinct from those of the amplified Est alpha 2(1), Est beta 2(1), or Est beta 1(1&2). An esterase with the same size EcoRI fragment as Est beta 1(3) was also present in Pel RR. This and restriction enzyme fragment analysis of C. quinquefasciatus field populations suggest that variability of the susceptible alleles may be lower than previously suggested. A non-amplified Est alpha with a unique EcoRI band was present in Pel RR. The previous esterase purification procedures may not have separated these amplified and non-amplified alleles. Hence, the small differences between the purified esterases from resistant strains may reflect mixtures of identical amplified alleles with different non-amplified alleles, which have significantly different k alpha values.

Animals↗
PubMed1995

Mosquito carboxylesterase Est alpha 2(1) (A2). Cloning and sequence of the full-length cDNA for a major insecticide resistance gene worldwide in the mosquito Culex quinquefasciatus.

Organophosphorus insecticide resistance in Culex mosquitoes is commonly caused by increased activity of one or more esterases. The commonest phenotype involves elevation of the esterases Est alpha 2 (A2) and Est beta 2 (B2). A cDNA encoding the Est alpha 2 esterase has now been isolated from a Sri Lankan insecticide-resistant mosquito (Culex quinquefasciatus, Say) expression library. In line with a recently suggested nomenclature system (Karunaratne, S. H. P. P. (1994) Characterization of Multiple Variants of Carboxylesterases Which Are Involved in Insecticide Resistance in the Mosquito Culex quinquefasciatus. Ph.D. thesis, University of London), as the first sequenced variant of this esterase, it is now referred to as Est alpha 2(1). The full-length cDNA of est alpha 2(1) codes for a 540-amino acid protein, which has high homology with other esterases and lipases and belongs to the serine or B-esterase enzyme family. The predicted secondary structure of Est alpha 2(1) is similar to the consensus secondary structure of proteins within the esterase/lipase family where the secondary and tertiary structures have been resolved. The level of identity (approximately 47% at the amino acid level) between the est alpha 2(1) and the various Culex est beta (B1 and B2) cDNA alleles that have been cloned and sequenced suggests that the two esterase loci are closely related and arose originally from duplication of a common ancestral gene. The lack of a distinct hydrophobic signal sequence for Est alpha 2(1) and two possible N-linked glycosylation sites, both situated close to the active site serine, suggest that it is a nonglycosylated protein that is not exported from the cell. Southern and dot blot analysis of genomic DNA from various insecticide-resistant and susceptible mosquito strains show that the est alpha 2(1) gene, like est beta 2(1), is amplified in resistant strains. The restriction fragment length polymorphism patterns, after probing Southern blots of EcoRI-digested genomic DNA with esta alpha 2(1) cDNA, show that the amplified and nonamplified est alpha alleles differ in the resistant and susceptible Sri Lankan mosquitoes.

Amino Acid Sequence↗
PubMed1995

The independent gene amplification of electrophoretically indistinguishable B esterases from the insecticide-resistant mosquito Culex quinquefasciatus.

Resistance to organophosphates in Culex mosquitoes is typically associated with increased activity of non-specific esterases. The commonest phenotype involves two elevated esterases, A2 and B2, while some strains have elevation of esterase B1 alone. Overexpression of the two B esterase electromorphs is due to gene amplification. Full-length cDNAs coding for amplified esterase B genes from a resistant Cuban strain (MRES, with amplified B1 esterase) and a Sri Lankan strain (PelRR, with amplified B2 esterase) of C. quinquefasciatus have been sequenced. In addition, a partial-length cDNA coding for a B esterase from an insecticide-susceptible Sri Lankan strain (PelSS) has been sequenced. All the nucleotide sequences and the inferred amino acid sequences show a high level of identify (> 95% at the nucleotide and amino acid level), confirming that they are an allelic series. The two B1 esterase nucleotide sequences (MRES and the previously published TEM-R [Mouches, Pauplin, Agarwal, Lemieux, Herzog, Abadon, Beyssat-Arnaouty, Hyrien, De Saint Vincent, Georghiou and Pasteur (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 2574-2578]) showed the lowest identity, and restriction-fragment-length-polymorphism analysis of the two strains was different. On the basis of these data we suggest that the two electrophoretically identical B1 esterase isoenzymes from California and Cuba have been amplified independently. Alternatively, if amplification has occurred only once, the original amplification has not occurred recently.

Amino Acid Sequence↗
PubMed1995

Efficacy of etofenprox against insecticide susceptible and resistant mosquito strains containing characterized resistance mechanisms.

Etofenprox is a non-ester pyrethroid insecticide with comparable toxicity and a similar mode of action to other pyrethroids. Cross-resistance studies on mosquitoes showed no effect of carboxylesterase, elevated esterase, altered acetylcholinesterase or glutathione S-transferase-based resistance mechanisms on etofenprox toxicity, when compared to standard susceptible strains of Anopheles and Culex. Cross-resistance to etofenprox occurred in a pyrethroid-resistant strain of Culex quinquefasciatus with both oxidase and 'kdr'-like resistance mechanisms. Dose-response data for susceptible mosquito strains suggest that, in standard W.H.O. susceptibility tests of adult mosquitoes, appropriate discriminating concentrations of etofenprox for detection of resistance would be 0.1% for Culex and 0.25% for Anopheles.

Animals↗
PubMed1995

Permethrin resistance in the head louse Pediculus capitis from Israel.

Head lice, Pediculus capitis, were collected from children aged 3-12 years in Maale Adumin, a town near Jerusalem, after reports of control failure with the pyrethroid insecticide permethrin. A total of 1516 children were examined: living lice and eggs were found on 12.1% of the children; or another 22.8% of the children only nits were found. Twice as many girls as boys (8.1% v 4%) were infested with lice and or nits. Head lice collected from infested children were exposed to permethrin impregnated filter-papers. Log time probit mortality (ltp) regression lines were calculated for mortality data and compared to ltp lines for a similar collection of head lice made in 1989. The regression lines for the two years were significantly different, with a 4-fold decrease in susceptibility at the LT50 level between 1989 and 1994. The slopes of the lines also suggested that the 1994 population was more heterogenous in its response to permethrin than the 1989 population. In contrast, a laboratory population of body lice (Pediculus humanus) tested with the same batch of permethrin-impregnated papers showed a slight but non-significant increase in susceptibility between 1989 and 1994. The results suggest that resistance to pyrethroids has developed rapidly among head lice since permethrin was introduced in 1991 as a pediculicide in Israel.

Animals↗
PubMed1995

Post-inoculation changes in enzyme activity of Aedes aegypti infected with Chikungunya virus.

Levels of acetylcholinesterase, non-specific esterases, glutathione-S-transferase and glucose-6-phosphate dehydrogenase in Aedes aegypti (L.) mosquitoes inoculated intrathoracally with Chikungunya virus were elevated, as compared to uninoculated control insects. A number of these enzymes are important in the insects defence mechanism against xenobiotics, such as pesticides. Malathion bioassays indicated a reduction in the susceptibility of experimentally injected insects with virus or virus-free inoculum, compared to non-inoculated controls. However, insects which were mock-inoculated (injected with no inoculum) showed a similar reduction in susceptibility suggesting that the observed effect was due to the mobilization of a defence reaction in the mosquitoes in response to injury during inoculation.

Acetylcholinesterase↗
PubMed1993

Characterization of a B-type esterase involved in insecticide resistance from the mosquito Culex quinquefasciatus.

The enzyme, esterase B2, involved in insecticide resistance has been purified and characterized from the mosquito Culex quinquefasciatus. The monomeric enzyme has an M(r) of 62000 and a pI of 5.0. This enzyme is compared with the esterase A2 previously characterized [Ketterman, Jayawardena and Hemingway (1992) Biochem. J. 287, 355-360]. The kinetic constants for interaction with several insecticides indicate, as for the esterase A2, that the B2 enzyme has a role in resistance. The rates and affinities of binding observed support the hypothesis that the role mainly is sequestration followed by the slow turnover of the insecticide. Although the B2 esterase appears to have a slightly higher rate of interaction with insecticides, the A2 has a much greater Vmax. with the xenobiotic substrates studied. The B2 esterase also appears to be present in the larvae to a lesser extent than the esterase A2.

Animals↗
PubMed1993

The selection and genetic analysis of esterase electromorphs in an organophosphate-resistant strain of Culex pipiens from Italy.

An Italian organophosphate-resistant strain of Culex pipiens (Lucca) was found to be polymorphic for elevated and nonelevated esterases. Selection for high esterase activity produced a strain homozygous for elevated esterases A2 and B2. Selection for low activity produced a strain homozygous for nonelevated esterases, A4i and B1i. Crossing experiments showed that A2 and B2 are coded by separate but closely linked genes, as are A4i and B1i. Results indicate that elevated A2 and nonelevated A4i are alleles of a single gene (Est-3 locus), as are elevated B2 and nonelevated B1i (Est-2 locus). Selection for electromorph variants gave four elevated A variants and three elevated B variants. These esterases were not selected in the field. In Lucca, A2 and B2 replaced A1, suggesting a selective advantage to the former over the latter in the presence of chlorpyrifos. It is hypothesized that the degree of amplification is an important factor in the selection of a particular esterase electromorph as a resistance mechanism and that migrating individuals with amplified genes could have an advantage when invading a population under selection.

Alleles↗
PubMed1993

A malaria control trial using insecticide-treated bed nets and targeted chemoprophylaxis in a rural area of The Gambia, west Africa. 3. Entomological characteristics of the study area.

Baseline entomological surveillance was carried out in a rural area of The Gambia during the rainy season in 1988, one year before the implementation of a malaria control programme using insecticide-impregnated nets and targeted chemoprophylaxis in villages with a primary health care (PHC) system. Mosquito collections took place in 6 pairs of settlements each with untreated bed nets; within each pair there was a large PHC village with a resident village health worker (VHW) and traditional birth attendant (TBA) and a smaller non-PHC village without either a VHW or a TBA. The most common vectors in the study area were Anopheles gambiae sensu stricto and, to a lesser extent, An. arabiensis. These mosquitoes were found in appreciable numbers for at least 4 months of the year (geometric mean/bedroom/night = 32.5, 95% confidence interval 18.2-57.3). Numbers of mosquitoes collected in PHC villages or non-PHC villages were not significantly different. Greater numbers of mosquitoes were found in villages closer to the River Gambia than in those further away. Evidence for DDT resistance due to elevated glutathione S-transferase activity was found in one of the 12 villages, but there was no evidence of resistance to organophosphate or carbamate insecticides as suggested by the low esterase levels and carbamate sensitive acetylcholinesterase.

Animals↗
PubMed1993

A malaria control trial using insecticide-treated bed nets and targeted chemoprophylaxis in a rural area of The Gambia, west Africa. 7. Impact of permethrin-impregnated bed nets on malaria vectors.

The impact of permethrin-impregnated bed nets on malaria vectors was studied in 6 pairs of villages during the rainy season in 1989. In each pair, the residents of one village had their nets treated whilst those of the other remained untreated. Routine collections of mosquitoes were made outdoors in the early evening using human-biting collections, and indoors with insecticide sprays, light traps and by searches under bed nets. Mosquitoes of the Anopheles gambiae complex, An. gambiae sensu stricto, An. arabiensis and An. melas, were present in large numbers for 5 months of the study period. These mosquitoes were susceptible to permethrin as judged by bioassay results. Outdoor human-biting rates in the early evening in communities with treated bed nets were similar to those in communities with untreated nets. In villages with treated bed nets most biting occurred outdoors in the early evening with little taking place under impregnated nets. The insecticidal activity of permethrin-impregnated bed nets, dipped by the local population, provided good individual protection against mosquitoes throughout the rainy season and bed nets remained effective even when washed up to 3 times. There was little to suggest that the use of insecticide-treated nets reduced the survival of mosquito populations in villages with impregnated nets. The absence of the expected village-wide effects of net impregnation may have resulted from the circulation of mosquitoes between villages with treated and untreated nets. The proportion of mosquitoes which fed on humans did not differ significantly between villages with treated and untreated nets.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗