Cholinesterase Activity In Tissues Research Articles

Can blood morphology, oxidative stress, and cholinesterase activity determine health status of pigeon Columba livia f. urbana?

Environmental studies in Northern Poland are example of the functioning of ecophysiological relationships under anthropogenic impact. The aim of our studies was to investigate sex-dependent effects on the alterations in the concentration of chemical elements in soil samples collected from habitats of feral pigeon Columba livia f. urbana from Northern Poland, as well as feathers, biomarkers of oxidative stress, antioxidant defense, and total cholinesterase activity in tissues (liver, kidney, brain). Concentration of Si, Zn, and Pb in feathers of pigeons was significant. The levels of Si and Zn were higher in feathers of females from non-polluted, while higher Pb levels were found only in females from polluted areas (p = 0.000). This was confirmed by MANOVA of biomarkers of antioxidant defense, elements concentration, and revealing the order of effects: tissue type > environment > sex. Erythrocytes of males living in polluted areas were more fragile to hemolytic agents resulting in a higher percentage of hemolyzed erythrocytes. The effects of polluted environment on the level of carbonyl derivatives of oxidatively modified proteins compared to the effects of sex were more pronounced in the case of kidney (p = 0.000) and hepatic tissues (p = 0.000). Polluted areas were associated with significant increase in SOD activity in the brain and hepatic tissues of pigeons (p = 0.000). Health status of feral pigeons is significantly different in conditions of environmental destabilization.

The Effects of Exposing Solea senegalensis to Microbeads with and Without Pesticides

Microplastics (MP) are contaminants able to cause adverse effects on organisms. MPs are capable to interact with other environmental contaminants, including pesticides, altering their toxicity. The objective of the study was to research the sublethal effects (enzymatic activity) of pesticides alone and in combination with MPs. Cholinesterase enzymes are used as biomarkers to determine and evaluate the effects produced in organisms after exposure to pollutants. This study showed the acetylcholinesterase (AChE) enzymatic activity in the tissue of Solea senegalensis exposed to two environmental pollutants, the insecticide chlorpyrifos (CPF) and antibacterial triclosan (TCS) with and without microplastics (MPs). Solea senegalensis was chosen because it is a species in high demand because of its high economic value in southern Europe, as well as the use of this species in ecotoxicology and its increasing use as sentinel species, which justify using it to assess biological effects of pollutants. Toxicity tests were performed in organisms exposed to concentrations of between 5 and 80 μg/L CPF and 0.1 and 0.4 mg/L TCS for 96 h. In addition, each test incorporated MPs that were added at different concentrations in order to evaluate their role as a possible enhancer of the effects caused by the previous pollutants. In the case of CPF, the head and muscle tissue cholinesterase activity was inhibited from a concentration of 5 μg/L both without and with MPs, and the AChE enzymatic activity for the treatment with MPs was approximately half the activity for the treatment without MPs in the tissues studied. Besides, TCS inhibited the cholinesterase activity at a concentration of 0.3 mg/L in the muscle of S. senegalensis. In contrast, no significant differences were observed in the TCS + MP treatment compared to the controls. These results showed the importance of studies in assessing the anticholinesterase effects of pesticides combined with microplastics due to the abundance of these contaminants in the marine environment and the role of cholinesterase activity (biomarker) in the neurotransmission of key physiological processes.

A bioassay on tissue cholinesterase activity of Rutilus kutum (Kamensky, 1901) exposed to some common pesticides in Iran.

The toxicological effects of three commonly used pesticides in Iran on the fries of Caspian kutum (Rutilus kutum, Kamensky, 1901) were investigated through determining 50.00% lethal concentration (LC50) 96hr and cholinesterase (ChE) inhibition. The LC50 96-hr of carbaryl, glyphosate, and malathion were equal to 11.69, 6.64, and 0.97 mg L-1, respectively, that were considered as harmful (10.00 - 100 mg L-1), toxic (1.00 - 10.00 mg L-1) and very toxic (< 1.00 mg L-1) compounds for this species. The exposure of fries to sub-lethal concentrations of the pesticides over 15 days indicated that the average of ChE activity in the head and trunk were 1086.89 ± 124.34 and 627.36 ± 99.60 mU min-1 per mg protein, respectively, with a significant difference relative to each other. There was a significant difference between fry exposed to all three pesticides and the control group in cholinesterase inhibition. The fries exposed to carbaryl (890.12 ± 28.08 mU min-1 per mg protein) and glyphosate (891.77 ± 31.61 mU min-1 per mg protein) showed lower ChE inhibition than those exposed to malathion (790.00 ± 58.14 mU min-1 per mg protein).

Comparative study of the cholinesterase activities in tissues of the round goby Neogobius melanostomus (Gobiidae) from different locations of the Black and Azov seas

Cholinesterase activity has been assayed in the liver, muscles, and blood serum of the round goby Neogobius melanostomus caught in the coastal waters of the Black and Azov seas differing in the degree of anthropogenic impact. The highest enzyme activity in the fish individuals from both seas is observed in the liver and the lowest is observed in the blood serum. The fish caught in the most polluted (by municipal and agricultural wastewaters) sites of both seas display a decrease in cholinesterase activity. The utility of cholinesterase activity as a biomarker in monitoring of the marine areas affected by anthropogenic pollution is discussed.

Cholinesterase activity in tissues of some crab species from the Japan Sea

The comparative study of the cholinesterase activity in some crab species was carried out for the first time with use of a set of thiocholine substrates. The substrate specificity was studied in stellar nerve, heart, and hemolymph of three crab species. The crab hemolymph was shown to be characterized by the highest enzyme activity. The enzyme from various crab organs has different structure o substrate specificity. Properties of crab enzymes was compared with acetylcholinesterase (AChE) of human blood erythrocytes, butyrylcholinesterase (BuChE) of horse blood serum, enzyme o squids and bivalve molluscs. The obtained data allow the conclusion to be made on differences in properties of enzymes both at the interspecies and at the tissue levels.

In vivo reactivation by oximes of inhibited blood, brain and peripheral tissue cholinesterase activity following exposure to nerve agents in guinea pigs

This study compared the ability of nine oximes (HI-6, HLö7, MMB-4, TMB-4, carboxime, ICD585, ICD692, ICD3805, and 2-PAM) to reactivate in vivo cholinesterase (ChE) in blood, brain, and peripheral tissues in guinea pigs intoxicated by one of four organophosphorus nerve agents. Two bis-pyridinium compounds without an oxime group, SAD128 and ICD4157, served as non-oxime controls. Animals were injected subcutaneously with 1.0× LD 50 of the nerve agents sarin, cyclosarin, VR or VX and treated intramuscularly 5 min later with one of these oximes. Toxic signs and lethality were monitored; tissue ChE activities were determined at 60 min after nerve agent. Some animals exposed to sarin or cyclosarin, with or without non-oxime treatment, died within 60 min; however, no animal treated with an oxime died. For VR or VX, all animals survived the 60 min after exposure, with or without non-oxime or oxime therapy. The four nerve agents caused differential degrees of inhibition in blood, brain regions and peripheral tissues. The tested oximes exhibited differential potency in reactivating nerve agent-inhibited ChE in various peripheral tissues, but did not affect ChE activity in the brain regions. There was no direct relation between blood and peripheral tissues in the reactivating efficacy of oxime treatments. ChE inhibited by sarin was the most susceptible to oxime reactivation while cyclosarin the least susceptible. There was no difference in the ChE reactivating potency between the dimethanesulfonate and dichloride salts of HI-6. MMB-4 significantly reactivated the ChE inhibited by these four nerve agents in blood and all three peripheral tissues of the guinea pig, and among all the oximes tested it was the most effective in vivo ChE reactivator against all four nerve agents.

Evaluation of nine oximes on in vivo reactivation of blood, brain, and tissue cholinesterase activity inhibited by organophosphorus nerve agents at lethal dose

The capability of several oximes (HI-6, HLö7, MMB-4, TMB-4, carboxime, ICD 585, ICD 692, ICD 3805, and 2-PAM) to reactivate in vivo AChE inhibited by the nerve agents sarin, cyclosarin, VX, or VR in blood, brain regions, and peripheral tissues in guinea pigs was examined and compared. Animals were injected subcutaneously with 1.0 LD50 of sarin, cyclosarin, VR, or VX, and treated intramuscularly 5 min later with one of these compounds. Toxic signs and lethality were monitored, and tissue AChE activities were determined at 60 min after nerve agent. The animals exposed to sarin or cyclosarin, alone or with non-oxime treatment, some died within 60 min; however, when treated with an oxime, no animal died. For VR or VX, all animals survived for 60 min after exposure, with or without non-oxime or oxime therapy. These nerve agents caused differential degrees of inhibition: in whole blood sarin = cyclosarin > VR = VX; in brain regions sarin > cyclosarin > VX > VR; and in peripheral tissues sarin > VX > cyclosarin > VR. These oximes exhibited differential potency in reactivating nerve agent-inhibited AChE in various peripheral tissues, but not AChE activity in the brain regions. There was no difference in the AChE reactivating potency between the dichloride and dimethanesulfonate salts of HI-6. AChE inhibited by sarin was the most and cyclosarin the least susceptible to oxime reactivation. Overall, MMB-4 appeared to be, among all oximes tested, the most effective in vivo AChE reactivator against the broadest spectrum of nerve agents.

Assessment of the Effect of Varying Soil Organic Matter Content on the Bioavailability of Malathion to the Common Nightcrawler, Lumbricus terrestris L.

This study investigated the effect of soil organic matter content on the bioavailability of malathion to the common nightcrawler, Lumbricus terrestris. Earthworms were exposed for 72 h to malathion on two soil types, 8% organic matter and 55% organic matter. Two different measures of bioavailability, malathion body burdens and tissue cholinesterase activities, were then measured in the malathion exposed animals. There were no significant differences in body burden or cholinesterase levels in L. terrestris exposed to malathion on soils with differing organic matter content. This suggests that absorption into organic matter is not a limiting factor of malathion bioavailability to earthworm species.

Time course of cholinesterase inhibition in adult rats treated acutely with carbaryl, carbofuran, formetanate, methomyl, methiocarb, oxamyl or propoxur

To compare the toxicity of seven N-methyl carbamates, time course profiles for brain and red blood cell (RBC) cholinesterase (ChE) inhibition were established for each. Adult, male, Long Evans rats (n=4-5 dose group) were dosed orally with either carbaryl (30 mg/kg in corn oil); carbofuran (0.5 mg/kg in corn oil); formetanate HCl (10 mg/kg in water); methomyl (3 mg/kg in water); methiocarb (25 mg/kg in corn oil); oxamyl (1 mg/kg in water); or propoxur (20 mg/kg in corn oil). This level of dosing produced at least 40% brain ChE inhibition. Brain and blood were taken from 0.5 to 24 h after dosing for analysis of ChE activity using two different methods: (1) a radiometric method which limits the amount of reactivation of ChE activity, and (2) a spectrophotometric method (Ellman method using traditional, unmodified conditions) which may encourage reactivation. The time of peak ChE inhibition was similar for all seven N-methyl carbamate pesticides: 0.5-1.0 h after dosing. By 24 h, brain and RBC ChE activity in all animals returned to normal. The spectrophotometric method underestimated ChE inhibition. Moreover, there was a strong, direct correlation between brain and RBC ChE activity (radiometric assay) for all seven compounds combined (r(2)=0.73, slope 1.1), while the spectrophotometric analysis of the same samples showed a poor correlation (r(2)=0.09). For formetanate, propoxur, methomyl, and methiocarb, brain and RBC ChE inhibitions were not different over time, but for carbaryl, carbofuran and oxamyl, the RBC ChE was slightly more inhibited than brain ChE. These data indicate (1) the radiometric method is superior for analyses of ChE activity in tissues from carbamate-treated animals (2) that animals treated with these N-methyl carbamate pesticides are affected rapidly, and recover rapidly, and (3) generally, assessment of RBC ChE is an accurate predictor of brain ChE inhibition for these seven pesticides.

A comparative study of the anticholinesterase activity of several antipsychotic agents

Drug-induced inhibition of plasma and tissue cholinesterase activity was evaluated in rats. The dopamine receptor antagonists haloperidol (HALO), chlorpromazine (CPZ), thioridazine (THIO), fluphenazine (FLU), clozapine (CLO) and sulpiride (SULP), used as neuroleptics, were tested. Two biochemical parameters were measured in vitro: the minimal effective concentration (MEC) for cholinesterase inhibition and the 50% inhibitory concentration (IC50). In addition, animals were tested for rotational activity after a unilateral intrastriatal injection of the drugs. The doses used for each drug were previously determined IC50s. After unilateral striatal drug injection, rats were challenged with intraperitoneal amphetamine injection in order to stimulate rotation. All drugs tested induced decreases in cholinesterase activity. Plasma MEC for THIO, FLU, HALO and CPZ were significantly lower than for CLO and SULP. In striatum, the MEC for TIO, CPZ and FLU was significantly lower than for HAL. According to plasma IC50, THIO, CPZ and CLO are potent cholinesterase inhibitors. CLO showed the lowest potency of cholinesterase inhibition in the striatum and THIO showed the highest potency in plasma and striatum. In conclusion, anticholinesterase activity is not related to D 2 receptor blockade or antipsychotic potency; and therefore the antipsychotic effects are not related to an increase in acetylcholine. All drugs induced similar contralateral rotation, except for CLO that was different from SULP and was not different from its control. Since equivalent cholinesterase inhibitory concentrations were used for all drugs and no differences in nigrostriatal behavioral effects were observed, these data suggest the participation of an important cholinergic component in this behavior. Therapeutically, the stronger the cholinesterase inhibition is, the more potent the cholinergic effects are and, consequently, the induction of extrapyramidal symptoms becomes more feasible.

Characterization of cholinesterase activity in tissues of the grass shrimp ( Palaemonetes pugio)

Inhibition of cholinesterases in estuarine organisms is a useful indicator of organophosphorus insecticide exposure. Recent research has indicated that more than one cholinesterase may be present in tissues of estuarine organisms and that these cholinesterases vary in their sensitivity to organophosphorus insecticides. Baseline cholinesterase activity for adult grass shrimp ( Palaemonetes pugio) was determined with four substrates (acetylthiocholine iodide, acetyl-β-methylthiocholine iodide, propionylthiocholine iodide, S-butyrylthiocholine iodide) and three cholinesterase inhibitors (eserine sulfate, tetraisopropyl pyrophosphoramide, 1,5-bis(4-allyldimethyl-ammoniumphenyl) pentan-3-one dibromide). Cholinesterase activity was the highest with acetylthiocholine iodide at 57.42 nmol/mg P/min and the lowest with acetyl-β-methylthiocholine iodide at 6.17 nmol/mg P/min. The four substrates tested were not inhibited by tetraisopropyl pyrophosphoramide. Eserine sulfate significantly inhibited cholinesterase activity against all substrates except S-butyrylthiocholine iodide. The results of substrate specificity and cholinesterase inhibition in grass shrimp indicate that acetylthiocholine iodide is the most appropriate substrate for assessing cholinesterase inhibition in grass shrimp.

Inhibitory effects of donepezil hydrochloride (E2020) on cholinesterase activity in brain and peripheral tissues of young and aged rats

Donepezil hydrochloride (donepezil: E2020: (±)-2-[(1-benzylpiperidin-4-yl)methyl]-5,6-dimethoxy-indan-1-one monohydrochloride)) is a centrally acting acetylcholinesterase inhibitor developed for the treatment of Alzheimer's disease. In the present study, its inhibitory effect on the activity of cholinesterase ex vivo was evaluated in the brain, plasma, erythrocytes, heart, small intestine, liver and pectoral muscle of young adult as well as aged rats, in comparison with that of tacrine (9-amino-1,2,3,4-tetrahydroacridine hydrochloride). In aged animals, cholinesterase activity in heart, small intestine and pectoral muscle was lower, whereas that in plasma and liver was higher than in young rats. Both groups showed the highest levels in the brain. Donepezil, at doses of 1.25, 2.5 and 5 mg/kg, p.o., inhibited brain, plasma, erythrocyte, liver and pectoral muscle cholinesterase activity in young rats in a dose-dependent manner but had less effect on cholinesterase activity in heart and small intestine. In aged animals, inhibition of cholinesterase activity in the brain, erythrocytes and pectoral muscle by donepezil was more potent than that in young animals. Tacrine, at doses of 5, 10 and 20 mg/kg, p.o., dose-dependently inhibited cholinesterase activity in all tissues of both young and aged animals, but most potently in heart, small intestine and liver. The inhibition of cholinesterase activity by tacrine in the brain, plasma, erythrocytes, heart and liver was more potent in aged rats than in tissues of young rats. Brain and plasma concentrations of unchanged donepezil and tacrine were measured in the same animals as used for the cholinesterase inhibition study. Brain and plasma concentrations of donepezil and tacrine were higher in aged than in young animals. It is concluded that the inhibitory effects of donepezil and tacrine on cholinesterase activity are greater in aged than in young rats, owing to differences in the tissue concentrations of these compounds between young and aged animals. It is also suggested that the effect of donepezil on cholinesterase activity is more tissue-selective than that of tacrine.

Acetylcholine content and tissue cholinesterase activity in rats with different motor activity

The content of acetylcholine and activity of cholinesterases in the heart and blood of 21, 30, 42, 49, 70, and 100-day-old rats were measured using standard biochemical methods. It is shown that functional activity of cholinergic system is lower in hypokinetic rats than in normokinetic age-matching controls.

Automated Instrument Analysis of Cholinesterase Activity in Tissues From Carbamate-Treated Animals: A Cautionary Note

Summary: When using a spectrophotometric method to measure cholinesterase activity in carbamate-treated tissues, precautions must be taken to limit the reactivation of the inhibited cholinesterase. Many testing laboratories use automated instruments to measure cholinesterase activity, instruments that usually employ a spectrophotometric method. To date, there has been no systematic investigation of the validity of the cholinesterase data obtained from carbamate-treated tissues using these automated instruments. The purpose of this study was to compare the cholinesterase data obtained using an automated analyzer with those obtained using a radiometric assay (i.e., the optimal method for unstable inhibitors). Using both an automated analyzer and the radiometric method, cholinesterase activity was measured in whole blood and brain tissue taken from rats treated with various dosages of carbaryl. Reactivation occurred when using the automated instrument. In the brain tissue, cholinesterase activity of the trea...

The effects of physostigmine on the electroretinogram in the beagle dog.

The purpose of the study was to correlate electroretinogram (ERG) parameters with increasing levels of plasma, erythrocyte and ocular tissue cholinesterase inhibition using the beagle dog as a model for human neurovisual toxicity. The anticholinesterase compound physostigmine was administered at various steady-state intravenous infusion rates based on pharmacokinetic estimates of plasma and red blood cell cholinesterase inhibition. The most sensitive parameter was the b-wave amplitude of the rod response, which was significantly depressed compared to pretreatment at all levels of acute cholinesterase depression. The overall maximal ERG response demonstrated a trend of declining a- and b-wave amplitudes, which corresponded with the increased levels of cholinesterase depression, but these differences were not significant. The depression of the electroretinogram rod and cone amplitudes appeared to parallel plasma cholinesterase inhibition more closely than erythrocyte cholinesterase activity. Ocular tissue cholinesterase activity was significantly depressed in the retina (70%), cornea (60%) and dorsal rectus extraocular muscle (46%). Electroretinography may be a useful physiological tool for evaluating the ocular toxicity of certain chemicals or pharmaceuticals associated with cholinesterase biomarker activity.

Differential effects of bromopride and domperidone on cholinesterase activity in rat tissues

Bromopride (BRO) and domperidone (DOMP) are dopamine D2 blocking agents used in gastroenterology clinics because of their anti emetic effect as well as their central and peripheral actions of increasing gastrointestinal motor activity. The rationale for these experiments was to compare BRO- and DOMP-effects on plasma, brain, and intestinal cholinesterase activity in vitro. BRO and DOMP effects on cholinesterase activity in plasma, striatum, duodenum and ileum of adult male rats were measured for drug concentrations ranging from 0.006 to 3.134 microM for BRO and from 0.006 to 125 microM for DOMP. The results demonstrate that both BRO and DOMP can inhibit cholinesterase activity in all tissues studied, with DOMP being more potent than BRO in plasma and intestinal tissues. These data suggest the existence of a cholinergic mechanism of action for these dopamine blocking agents.

Differential effects of bromopride and domperidone on cholinesterase activity in rat tissues

Bromopride (BRO) and domperidone (DOMP) are dopamine D2 blocking agents used in gastroenterology clinics because of their anti emetic effect as well as their central and peripheral actions of increasing gastrointestinal motor activity. The rationale for these experiments was to compare BRO- and DOMP-effects on plama brain, and intestinal cholinesterase activity in vitro. BRO- and DOMP effects on cholinesterase activity in plasma, striatum, duodenum and ileum of adult male rats were measured for drug concentrations ranging from 0.006 to 3.134 μM for BRO and from 0.006 to 125 μM for DOMP. The results demonstrate that both BRO and DOMP can inhibit cholinesterase activity in all tissues studied, with DOMP being more potent than BRO in plasma and intestinal tissues. These data suggest the existence of a cholinergic mechanism of action for these dopamine blocking agents.

A Modified Spectrophotometric Method Appropriate for Measuring Cholinesterase Activity in Tissue from Carbaryl-Treated Animals

A Modified Spectrophotometric Method Appropriate for Measuring Cholinesterase Activity in Tissue from Carbaryl-Treated Animals. Nostrandt, A. C., Duncan, J. A., and Padilla, S. (1993). Fundam. Appl. Toxicol. 21, 196-203. Inhibited cholinesterase in tissues of animals exposed to carbamate pesticides is known to reactivate readily, presenting considerable problems in the accurate assessment of cholinesterase activity in these tissues. Decarbamylation of cholinesterase is favored when the tissue samples are diluted and/or are incubated for an extended time. The present study was performed to identify modifications of the commonly used spectrophotometric assay for cholinesterase activity that would minimize spontaneous reactivation of enzyme activity. Those modifications included preincubation of concentrated tissue with concentrated chromogen (i.e., DTNB), dilution to final reaction volume immediately before measurement, and measurement of cholinesterase over a short period of time (5-10 min). The Ellman assay with and without modifications was performed using a microtiter plate reader on tissues from carbaryl-treated rats: undiluted plasma, diluted erythrocytes (1:25), minimally diluted erythrocytes (1:2), diluted brain (1:100), or minimally diluted brain (1:2). The results were compared to cholinesterase activities obtained using a radiometric method which employs minimally diluted tissue and short incubation times. The degree of cholinesterase inhibition for undiluted or minimally diluted tissue assayed by the modified method agreed with those obtained using the radiometric method. Even if the tissues were diluted immediately before assay, however, significant reactivation occurred by the time the first measurements were made by the conventional method. Furthermore, significant spontaneous reactivation may still occur using the modified method if the assay is run for more than 10 min. Use of this modified Ellman method will enable more accurate estimation of in vivo cholinesterase activity in animals treated with carbamates.

A Modified Spectrophotometric Method Appropriate for Measuring Cholinesterase Activity in Tissue from Carbaryl-Treated Animals

A Modified Spectrophotometric Method Appropriate for Measuring Cholinesterase Activity in Tissue from Carbaryl-Treated Animals

Cresylbenzodioxaphosphorin oxide pretreatment alters soman-induced toxicity and inhibition of tissue cholinesterase activity of the rat

The toxicity of soman was investigated in the rat with and without pretreatment with Cresylbenzodioxa-phosphorin oxide (CBDP). Without pretreatment, the 24-h LD 50 for soman was 118.2 μg/kg s.c., and soman inhibited Carboxylesterase (CaE) activity in plasma (ED 50 of 55 μg/kg) and cholinesterase (ChE) activity in brain regions (ED 50 values of 65–105 μg/kg) in a dose-related manner. With pretreatment, the 24-h LD 50 for soman was reduced by approximately 6-fold and 8-fold (by 1.0 mg/kg and 16.0 mg/kg of CBDP, respectively), and the ED 50 values for soman-induced inhibition of ChE activity in brain regions were reduced by approximately 10-fold (by 1.0 mg/kg of CBDP). The dose-dependent severity of soman intoxication varied widely in rats treated with soman alone but not in CBDP-pretreated rats, and the ED 50 for the occurrence of signs of soman intoxication was reduced approximately 7-fold following CBDP (1.0 mg/kg) pretreatment. These data support the hypothesis that CBDP pretreatment effectively blocks tissue CaE sites which serve to detoxify soman, thus potentiating both the soman-induced inhibition of ChE in the CNS and the lethality of soman.