Rapid Method for Measurement of Acetylcholinesterase Activity

Antokhin, A. M.; Gainullina, E. T.; Ryzhikov, S. B.; Taranchenko, V. F.; Yavaeva, D. K.

doi:10.1007/s10517-009-0436-z

Cited by 5 publications

(4 citation statements)

References 3 publications

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“…Blood acetylcholinesterase (BChE) activity was measured on posttreatment days one and seven, by the method described by Antokhin et al [44]. In the presence of BChE, the hydrolysis of Ach and production of acetic acid decreases the pH of the reaction mixture.…”

Section: Blood Acetylcholinesterase Activitymentioning

confidence: 99%

Commercial Formulation of Chlorpyrifos Alters Neurological Behaviors and Fertility

Kudavidanage

Dissanayake

Keerthirathna

et al. 2020

Biology

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Pesticides are known to result in toxic insult. We aimed to evaluate Judo 40, the commercial formulation of chlorpyrifos on the neurological activities, fertility, and hormone levels of male rats. Male Wistar rats were treated orally with 1 mL of 20 or 50 mg/kg Judo 40. The doses were administered four times, twice a day. Sexual and exploratory behavior indices, fertility indices, serum androgen levels, blood acetylcholinesterase (BChE) levels, and neurological and muscular effects were evaluated. Serum testosterone and luteinizing hormone were significantly reduced in the rats receiving 50 mg/kg Judo 40. A reduction in viable implantation sites and live pups born were evident in the female rats mated with the male rats treated with the highest dose. Similarly, in the rats treated with the highest dose of Judo 40, a significant reduction in plasma BChE enzyme was observed. According to the results, prolonged Judo 40 exposure can cause impairment of the neurological alterations and sex hormones leading to impaired fertility. Therefore, chemical handlers should be educated on protection and risk minimization.Biology 2020, 9, 49 2 of 15 environment, chemical formulations of CPFs such as Judo 40 will undergo evaporation, hydrolysis, oxidation, photolysis, and microbial metabolism [15]. Owing to active soil absorption and the low water dissolubility of CPFs, this can result in the contamination of percolated water, runoff water, and sediments [16], leading to a high accumulation of CPF residues, thus posing acute and chronic health risks to consumers [17].It has been documented that the occupational exposure of humans to pesticides, including CPFs, are high [18]. However, in developing countries, small-scale farmers are continuously exposed to pesticides through mixing, filling, and spraying using backpack sprinklers [19]. They are further at a high risk due to the limited usage of protective gear and lack of awareness on safety [20]. In Sri Lanka, farmers are inclined to use about 30%-40% higher quantities of chemical formulations of CPF than the dose levels recommended by the manufacturers [21]. Growing evidence also indicates that CPF exposure can result in fetal developmental problems, including issues with the weight and head circumference of newborn babies [22], and high incidences of prostate and lung cancers in adults [23]. Alvarez et al. [24] reported that the soil can recover up to 25 ppm of CPF and the percentage decrease with increasing of the initial concentration [25]. The use of commercial formulations of pesticides can aggravate the toxic effects more than use of the pure active ingredient [26].The active ingredient of Judo 40, CPF, is principally responsible for the accumulation of acetylcholine (ACh) as a result of inhibition of acetylcholinesterase (AChE) action in animal tissues, and is highly toxic to many animals including copepods, amphipods [27], fish [28], birds, and mammals [29]. It was reported that chronic exposure to 30 days of chronic doses (5 and 10 mg/kg) of CPF led to a decrease in spe...

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Section: Blood Acetylcholinesterase Activitymentioning

confidence: 99%

Commercial Formulation of Chlorpyrifos Alters Neurological Behaviors and Fertility

Kudavidanage

Dissanayake

Keerthirathna

et al. 2020

Biology

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“…Thioflavine T was chosen for the development of this biosensor because of the high fluorescence inten sity of its complex with the recombinant AChE of human erythrocytes, which is higher than the fluores cence intensity of the initial thioflavine T at λ ~490 nm upon excitation with radiation at λ = 448 nm by a fac tor of 1000 [34]. A considerable increase in the fluo rescence intensity of thioflavine T in the presence of the AChE of human erythrocytes was observed by Antokhin et al [35]. Scheme 2 shows the structural formula of thioflavine T. The fluorescence intensity of the discussed optical biosensor under the action of the insecticide (~50 nM) diethyl (3,5,6 trichloropyridin 2 yl) phosphate or dichlophos decreased by 50% rela tive to the initial fluorescence intensity of the optical biosensor [33].…”

Section: Analysis Of Promising Trends In the Development Of Up To Datmentioning

confidence: 99%

Biosensors as tools of environmental monitoring for organophosphorus nerve agents

Gainullina¹,

Gulikova²,

Korneev³

et al. 2015

J Anal Chem

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Current advances in the development of tools of environmental monitoring for anticholinesterase agents are considered. The aim of this analysis was to substantiate the most promising trends in the develop ment of an up to date biosensor based on cholinesterase inhibition with an optimum sensitivity-analytical signal formation time relation for the monitoring of atmospheric air for organophosphorus warfare agents and insecticides.

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“…The three-dimensional structure of acetylcholinesterase (Figure ) reveals a deep narrow gorge that contains two ligand binding sites: a peripheral site (P-site) near the gorge entrance and an acylation site (A-site) at the base of the gorge . Thioflavin T (ThT), a fluorescent probe widely used as a test for amyloid fibril formation, − binds selectively to the AChE peripheral site, wherein the ThT fluorescence intensity increases significantly. − The proved proportionality of the dye’s fluorescence intensity to AChE activity led to the elaboration of a rapid fluorescent method for determination of AChE activity using ThT . Furthermore, it was shown that with binding of another ligand to the A-site in a ternary complex and initiation of a change in the local AChE conformation at the peripheral site, the enhanced fluorescence of ThT is quenched in 1.5 to 4-fold. ,, This fact allows ThT to be used as a reporter for ligand reactions at the AChE acylation site.…”

Section: Introductionmentioning

confidence: 99%

“…14−16 The proved proportionality of the dye's fluorescence intensity to AChE activity led to the elaboration of a rapid fluorescent method for determination of AChE activity using ThT. 17 Furthermore, it was shown that with binding of another ligand to the A-site in a ternary complex and initiation of a change in the local AChE conformation at the peripheral site, the enhanced fluorescence of ThT is quenched in 1.5 to 4fold. 10,16,18 This fact allows ThT to be used as a reporter for ligand reactions at the AChE acylation site.…”

Section: Introductionmentioning

confidence: 99%

Thioflavin T Interaction with Acetylcholinesterase: New Evidence of 1:1 Binding Stoichiometry Obtained with Samples Prepared by Equilibrium Microdialysis

Sulatskaya

Rychkov

Sulatsky

et al. 2018

ACS Chem. Neurosci.

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The aim of the present work was investigation of the fluorescent dye thioflavin T (ThT) binding to acetylcholinesterase (AChE). ThT is an effective test for protease activity, as well as a probe for amyloid fibril formation. Despite the extended and active investigation of ThT-AChE binding, there is still no common view on the stoichiometry of this interaction. In particular, there is a hypothesis explaining the spectral properties of bound to AChE dye and high quantum yield of its fluorescence by formation of dimers or excimers of ThT. In order to confirm or deny this hypothesis, we proposed a new experimental approach for examination of ThT-AChE interaction based on spectroscopic investigation of samples prepared by equilibrium microdialysis. This approach allowed us to prove 1/1 ThT/AChE binding stoichiometry. The increase of ThT fluorescence quantum yield and lifetime accompanying its binding to AChE can be explained by the molecular rotor nature of this dye. Together with the coincidence of the positions of free and AChE-bound ThT fluorescence spectra, the obtained results prove the groundlessness of the hypotheses about ThT aggregation while binding to AChE. The model of ThT localization in the active site of AChE was proposed by using molecular docking simulations. These results also allowed us to suggest the key role of aromatic residues in ThT-AChE interaction, as observed for some amyloid fibrils.

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Rapid Method for Measurement of Acetylcholinesterase Activity

Cited by 5 publications

References 3 publications

Commercial Formulation of Chlorpyrifos Alters Neurological Behaviors and Fertility

Commercial Formulation of Chlorpyrifos Alters Neurological Behaviors and Fertility

Biosensors as tools of environmental monitoring for organophosphorus nerve agents

Thioflavin T Interaction with Acetylcholinesterase: New Evidence of 1:1 Binding Stoichiometry Obtained with Samples Prepared by Equilibrium Microdialysis

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