scholarly journals Purification and characterization of a carboxylesterase involved in insecticide resistance from the mosquito Culex quinquefasciatus

1992 ◽  
Vol 287 (2) ◽  
pp. 355-360 ◽  
Author(s):  
A J Ketterman ◽  
K G Jayawardena ◽  
J Hemingway
Keyword(s):  
Insecticide Resistance ◽  
Culex Quinquefasciatus ◽  
Minor Role ◽  
Organophosphate Insecticide ◽  
Purification And Characterization ◽  
Medium Chain Length ◽  
A Minor ◽  
Slow Turnover

A carboxylesterase (EC 3.1.1.1) involved in organophosphate insecticide resistance has been purified and characterized from the mosquito Culex quinquefasciatus. The monomeric enzyme has M(r) of 67,000 and a pI of 5.2. It hydrolysed medium-chain-length mono- and di-acylglycerols in addition to xenobiotic esters. Kinetic constants determined for four insecticides, temephos, chlorpyrifos, fenitrothion and propoxur indicate the rates of acylation and the affinities of binding of the insecticides to this carboxylesterase are important. This supports the major role of the A2 carboxylesterase is the sequestration of the insecticide with a minor role in the slow turnover of the insecticide.

scholarly journals Aldehyde dehydrogenase is essential for both adult and larval ethanol resistance in Drosophila melanogaster

2006 ◽  
Vol 87 (2) ◽  
pp. 87-92 ◽  
Author(s):  
JAMES D. FRY ◽  
MOLLY SAWEIKIS
Keyword(s):  
Aldehyde Dehydrogenase ◽  
Ethanol Metabolism ◽  
Minor Role ◽  
Ethanol Sensitivity ◽  
Fruit Breeding ◽  
Structural Locus ◽  
Ethanol Resistance ◽  
A Minor

The enzyme aldehyde dehydrogenase (ALDH) is essential for ethanol metabolism in mammals, converting the highly toxic intermediate acetaldehyde to acetate. The role of ALDH in Drosophila has been debated, with some authors arguing that, at least in larvae, acetaldehyde detoxification is carried out mainly by alcohol dehydrogenase (ADH), the enzyme responsible for converting ethanol to acetaldehyde. Here, we report the creation and characterization of four null mutants of Aldh, the putative structural locus for ALDH. Aldh null larvae and adults are poisoned by ethanol concentrations easily tolerated by wild-types; their ethanol sensitivity is in fact comparable to that of Adh nulls. The results refute the view that ALDH plays only a minor role in ethanol detoxification in larvae, and suggest that Aldh and Adh may be equally important players in the evolution of ethanol resistance in fruit-breeding Drosophila.

scholarly journals Occurrence of L1014F and L1014S mutations in insecticide resistant Culex quinquefasciatus from filariasis endemic districts of West Bengal, India

2022 ◽  
Vol 16 (1) ◽  
pp. e0010000
Author(s):  
Priyanka Rai ◽  
Dhiraj Saha
Keyword(s):  
Culex Quinquefasciatus ◽  
West Bengal ◽  
Economic Status ◽  
Minor Role ◽  
Resistance Development ◽  
Vector Population ◽  
A Minor ◽  
First Time

Introduction Lymphatic filariasis causes long term morbidity and hampers the socio-economic status. Apart from the available treatments and medication, control of vector population Culex quinquefasciatus Say through the use of chemical insecticides is a widely applied strategy. However, the unrestrained application of these insecticides over many decades has led to resistance development in the vectors. Methods In order to determine the insecticide susceptibility/resistance status of Cx. quinquefasciatus from two filariasis endemic districts of West Bengal, India, wild mosquito populations were collected and assayed against six different insecticides and presence of L1014F; L1014S kdr mutations in the voltage-gated sodium channel gene was also screened along with the use of synergists to evaluate the role of major detoxifying enzymes in resistance development. Results The collected mosquito populations showed severe resistance to insecticides and the two synergists used–PBO (piperonyl butoxide) and TPP (triphenyl phosphate), were unable to restore the susceptibility status of the vector thereupon pointing towards a minor role of metabolic enzymes. kdr mutations were present in the studied populations in varying percent with higher L1014F frequency indicating its association with the observed resistance to pyrethroids and DDT. This study reports L1014S mutation in Cx. quinquefasciatus for the first time.

scholarly journals Tudor Nuclease Genes and Programmed DNA Rearrangements in Tetrahymena thermophila

2007 ◽  
Vol 6 (10) ◽  
pp. 1795-1804 ◽  
Author(s):  
Rachel A. Howard-Till ◽  
Meng-Chao Yao
Keyword(s):  
Sexual Reproduction ◽  
Small Rna ◽  
Tetrahymena Thermophila ◽  
Minor Role ◽  
Dna Deletion ◽  
Genome Defense ◽  
Rna Pathways ◽  
A Minor

ABSTRACT Proteins containing a Tudor domain and domains homologous to staphylococcal nucleases are found in a number of eukaryotes. These “Tudor nucleases” have been found to be associated with the RNA-induced silencing complex (A. A. Caudy, R. F. Ketting, S. M. Hammond, A. M. Denli, A. M. Bathoorn, B. B. Tops, J. M. Silva, M. M. Myers, G. J. Hannon, and R. H. Plasterk, Nature 425:411-414, 2003). We have identified two Tudor nuclease gene homologs, TTN1 and TTN2, in the ciliate Tetrahymena thermophila, which has two distinct small-RNA pathways. Characterization of single and double KOs of TTN1 and TTN2 shows that neither of these genes is essential for growth or sexual reproduction. Progeny of TTN2 KOs and double knockouts occasionally show minor defects in the small-RNA-guided process of DNA deletion but appear to be normal in hairpin RNA-induced gene silencing, suggesting that Tudor nucleases play only a minor role in RNA interference in Tetrahymena. Previous studies of Tetrahymena have shown that inserted copies of the neo gene from Escherichia coli are often deleted from the developing macronucleus during sexual reproduction (Y. Liu, X. Song, M. A. Gorovsky, and K. M. Karrer, Eukaryot. Cell 4:421-431, 2005; M. C. Yao, P. Fuller, and X. Xi, Science 300:1581-1584, 2003). This transgene deletion phenomenon is hypothesized to be a form of genome defense. Analysis of the Tudor nuclease mutants revealed exceptionally high rates of deletion of the neo transgene at the TTN2 locus but no deletion at the TTN1 locus. When present in the same genome, however, the neo gene is deleted at high rates even at the TTN1 locus, further supporting a role for trans-acting RNA in this process. This deletion is not affected by the presence of the same sequence in the macronucleus, thus providing a counterargument for the role of the macronuclear genome in specifying all sequences for deletion.

scholarly journals σB Modulates Virulence Determinant Expression and Stress Resistance: Characterization of a Functional rsbU Strain Derived from Staphylococcus aureus 8325-4

2002 ◽  
Vol 184 (19) ◽  
pp. 5457-5467 ◽  
Author(s):  
Malcolm J. Horsburgh ◽  
Joanne L. Aish ◽  
Ian J. White ◽  
Les Shaw ◽  
James K. Lithgow ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Minor Role ◽  
Gene Encoding ◽  
Phenotypic Properties ◽  
Relative Level ◽  
General Stress Response ◽  
A Minor ◽  
Derivatives Of

ABSTRACT The accessory sigma factor σB controls a general stress response that is thought to be important for Staphylococcus aureus survival and may contribute to virulence. The strain of choice for genetic studies, 8325-4, carries a small deletion in rsbU, which encodes a positive regulator of σB activity. Consequently, to enable the role of σB in virulence to be addressed, we constructed an rsbU + derivative, SH1000, using a method that does not leave behind an antibiotic resistance marker. The phenotypic properties of SH1000 (8325-4 rsbU +) were characterized and compared to those of 8325-4, the rsbU mutant, parent strain. A recognition site for σB was located in the promoter region of katA, the gene encoding the sole catalase of S. aureus, by primer extension analysis. However, catalase expression and activity were similar in SH1000 (8325-4 rsbU +), suggesting that this promoter may have a minor role in catalase expression under normal conditions. Restoration of σB activity in SH1000 (8325-4 rsbU +) resulted in a marked decrease in the levels of the exoproteins SspA and Hla, and this is likely to be mediated by reduced expression of agr in this strain. By using Western blotting and a sarA-lacZ reporter assay, the levels of SarA were found to be similar in strains 8325-4 and SH1000 (8325-4 rsbU +) and sigB mutant derivatives of these strains. This finding contrasts with previous reports that suggested that SarA expression levels are altered when they are measured transcriptionally. Inactivation of sarA in each of these strains resulted in an expected decrease in agr expression; however, the relative level of agr in SH1000 (8325-4 rsbU +) remained less than the relative levels in 8325-4 and the sigB mutant derivatives. We suggest that SarA is not likely to be the effector in the overall σB-mediated effect on agr expression.

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

1993 ◽  
Vol 294 (2) ◽  
pp. 575-579 ◽  
Author(s):  
S H P P Karunaratne ◽  
K G Jayawardena ◽  
J Hemingway ◽  
A J Ketterman
Keyword(s):  
Insecticide Resistance ◽  
Culex Quinquefasciatus ◽  
Kinetic Constants ◽  
Slow Turnover

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.

Determination of the role of elevated B2 esterase in insecticide resistance in Culex quinquefasciatus (Diptera: Culicidae) from studies on the purified enzyme

1994 ◽  
Vol 84 (1) ◽  
pp. 39-43 ◽  
Author(s):  
K.G.I. Jayawardena ◽  
S.H.P.P. Karunaratne ◽  
A.J. Ketterman ◽  
J. Hemingway
Keyword(s):  
Culex Quinquefasciatus ◽  
Turnover Rate ◽  
Specific Activity ◽  
Sri Lankan ◽  
Organophosphorus Insecticides ◽  
Purification And Characterization ◽  
Sds Page

AbstractA method for the purification of elevated esterase B2 from the Sri Lankan Pel RR strain of Culex quinquefasciatus (Say) has been developed, using sequential column chromatography. The pure enzyme is stable, with no decrease in specific activity after several months, if stored in the presence of 25 mM dithiothreitol and 50% glycerol at –20°C. The enzyme has a pl of 5 and an estimated molecular weight for the monomeric enzyme of 63 kD from SDS-PAGE. In all Culex strains with elevated B2, another esterase (A2) is always co-elevated. The strains with these esterases are resistant to a broad range of organophosphorus insecticides. The previous purification and characterization of A2 from Pel RR indicated that the role of this enzyme was primarily to sequester rather than metabolize organophosphates. Purified B2, rapidly binds malaoxon, the activated form of malathion, but not malathion itself. The turnover rate of bound malaoxon is slow, suggesting that the role of this esterase in malathion resistance is also sequestration. The similar bimolecular rate constants for both A2 and B2 with malaoxon suggest both esterases are important in resistance, which may explain the almost complete linkage disequilibrium that the elevated forms of these two esterases are maintained in when under pesticide selection pressure.

scholarly journals Isolation and characterization of barley (Hordeum vulgare) extracellular vesicles and their role in RNAi-based crop protection

2021 ◽  
Author(s):  
T Schlemmer ◽  
L Weipert ◽  
C Preußer ◽  
M Hardt ◽  
A Möbus ◽  
...  
Keyword(s):  
Hordeum Vulgare ◽  
Gene Silencing ◽  
Extracellular Vesicles ◽  
Crop Protection ◽  
Minor Role ◽  
Transgenic Expression ◽  
Isolation And Characterization ◽  
A Minor

AbstractThe demonstration that spray-induced gene silencing (SIGS) can confer strong disease resistance bypassing the laborious and time-consuming transgenic expression of double-stranded (ds)RNA to induce gene silencing of pathogenic targets was groundbreaking. However, future field applications will require fundamental mechanistic knowledge on dsRNA uptake, processing, and its transfer. There is increasing evidence that extracellular vesicles (EVs) mediate the transfer of transgene-derived small interfering (si)RNAs in host-induced gene silencing (HIGS) applications. Here, we examined the role of EVs regarding the translocation of sprayed dsRNA from barley (Hordeum vulgare) to the target fungusFusarium graminearum. We found barley EVs with 156 nm in size containing predominantly 21 and 19 nucleotide (nt) siRNAs starting with a 5’-terminal Adenine. Notably, barley EVs contain less siRNA compared to EVs isolated from transgenic HIGS Arabidopsis plants. Together our results further underpin mechanistic differences between HIGS and SIGS applications and a minor role of EVs in SIGS.

scholarly journals The genic view of plant speciation: recent progress and emerging questions

2008 ◽  
Vol 363 (1506) ◽  
pp. 3023-3036 ◽  
Author(s):  
Christian Lexer ◽  
Alex Widmer
Keyword(s):  
Gene Networks ◽  
Recent Progress ◽  
Chromosomal Rearrangements ◽  
Future Research ◽  
Minor Role ◽  
Genetic Studies ◽  
A Minor ◽  
Species Isolation

The genic view of the process of speciation is based on the notion that species isolation may be achieved by a modest number of genes. Although great strides have been made to characterize ‘speciation genes’ in some groups of animals, little is known about the nature of genic barriers to gene flow in plants. We review recent progress in the characterization of genic species barriers in plants with a focus on five ‘model’ genera: Mimulus (monkey flowers); Iris (irises); Helianthus (sunflowers); Silene (campions); and Populus (poplars, aspens, cottonwoods). The study species in all five genera are diploid in terms of meiotic behaviour, and chromosomal rearrangements are assumed to play a minor role in species isolation, with the exception of Helianthus for which data on the relative roles of chromosomal and genic isolation factors are available. Our review identifies the following key topics as being of special interest for future research: the role of intraspecific variation in speciation; the detection of balancing versus directional selection in speciation genetic studies; the timing of fixation of alleles of major versus minor effects during plant speciation; the likelihood of adaptive trait introgression; and the identification and characterization of speciation genes and speciation gene networks.

The Effect of Path Length and Display Size on Memory for Spatial Information

2012 ◽  
Vol 59 (3) ◽  
pp. 147-152 ◽  
Author(s):  
Katherine Guérard ◽  
Sébastien Tremblay
Keyword(s):  
Path Length ◽  
Potential Role ◽  
Spatial Information ◽  
Recall Performance ◽  
Minor Role ◽  
Length Effect ◽  
Serial Memory ◽  
Fixed Reference ◽  
A Minor

In serial memory for spatial information, some studies showed that recall performance suffers when the distance between successive locations increases relatively to the size of the display in which they are presented (the path length effect; e.g., Parmentier et al., 2005) but not when distance is increased by enlarging the size of the display (e.g., Smyth & Scholey, 1994). In the present study, we examined the effect of varying the absolute and relative distance between to-be-remembered items on memory for spatial information. We manipulated path length using small (15″) and large (64″) screens within the same design. In two experiments, we showed that distance was disruptive mainly when it is varied relatively to a fixed reference frame, though increasing the size of the display also had a small deleterious effect on recall. The insertion of a retention interval did not influence these effects, suggesting that rehearsal plays a minor role in mediating the effects of distance on serial spatial memory. We discuss the potential role of perceptual organization in light of the pattern of results.

Effects of Myo-inositol Alone and in Combination with Seleno-Lmethionine on Cadmium-Induced Testicular Damage in Mice

2019 ◽  
Vol 12 (4) ◽  
pp. 311-323 ◽  
Author(s):  
Salvatore Benvenga ◽  
Antonio Micali ◽  
Giovanni Pallio ◽  
Roberto Vita ◽  
Consuelo Malta ◽  
...  
Keyword(s):  
Structural Changes ◽  
Natural Antioxidants ◽  
Minor Role ◽  
Positive Role ◽  
Testosterone Levels ◽  
Tnf Α ◽  
Testicular Lesions ◽  
A Minor ◽  
Immunohistochemical Analyses

Background: Cadmium (Cd) impairs gametogenesis and damages the blood-testis barrier. Objective: As the primary mechanism of Cd-induced damage is oxidative stress, the effects of two natural antioxidants, myo-inositol (MI) and seleno-L-methionine (Se), were evaluated in mice testes. Methods: Eighty-four male C57 BL/6J mice were divided into twelve groups: 0.9% NaCl (vehicle; 1 ml/kg/day i.p.); Se (0.2 mg/kg/day per os); Se (0.4 mg/kg/day per os); MI (360 mg/kg/day per os); MI plus Se (0.2 mg/kg/day); MI plus Se (0.4 mg/kg/day); CdCl2 (2 mg/kg/day i.p.) plus vehicle; CdCl2 plus MI; CdCl2 plus Se (0.2 mg/kg/day); CdCl2 plus Se (0.4 mg/kg/day); CdCl2 plus MI plus Se (0.2 mg/kg/day); and CdCl2 plus MI plus Se (0.4 mg/kg/day). After 14 days, testes were processed for biochemical, structural and immunohistochemical analyses. Results: CdCl2 increased iNOS and TNF-α expression and Malondialdehyde (MDA) levels, lowered glutathione (GSH) and testosterone, induced testicular lesions, and almost eliminated claudin-11 immunoreactivity. Se administration at 0.2 or 0.4 mg/kg significantly reduced iNOS and TNF-α expression, maintained GSH, MDA and testosterone levels, structural changes and low claudin-11 immunoreactivity. MI alone or associated with Se at 0.2 or 0.4 mg/kg significantly reduced iNOS and TNF-α expression and MDA levels, increased GSH and testosterone levels, ameliorated structural organization and increased claudin-11 patches number. Conclusion: We demonstrated a protective effect of MI, a minor role of Se and an evident positive role of the association between MI and Se on Cd-induced damages of the testis. MI alone or associated with Se might protect testes in subjects exposed to toxicants, at least to those with behavior similar to Cd.

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