Determination of the Inhibitory Effect of 2,2,4-trimethyl-2,3-dihydro-1-Hbenzo[b][1,4]diazepine on Acetylcholinesterase Enzyme Activity

The pineal gland releases melatonin to regulate our body's circadian rhythm based on light and dark cycles. The pinealectomy (PINX) model is an experimental approach employed to investigate the potential impact of melatonin on various tissues and pathologies. In this study, acetylcholinesterase (AChE) enzyme activity levels, oxidative stress parameters, histopathological findings, and serum melatonin levels in rat brain tissue were evaluated following pinealectomy. 24 male Sprague Dawley rats were randomly divided into groups: control, Sham-Pinealectomy (SHAM), and PINX. Brain tissue samples were taken at the end of a 50-day experimental period to determine the parameters of AChE, glutathione s-transferase (GST), carboxylesterase (Ces) enzyme activities, and malondialdehyde (MDA), reduced glutathione (GSH) levels spectrophotometrically. Moreover, serum melatonin levels were measured, and tissues underwent standard histological analysis to determine the histopathological damage score. In this study, we found that the PINX group had decreased AChE and Ces enzyme activity, increased MDA, decreased GSH levels, and no change in GST enzyme activity. A relative decrease in serum melatonin levels was also observed in the PINX group. In the light microscopic examination of the brain tissue of pinealectomy rats, it was observed that the eosinophilic staining intensity increased, heterochromatic/pycnotic-looking neuron nuclei were prominent in the cortex layers and hippocampus, and perineural edematous areas were abundant. Excessive perineuronal edema, cytoplasmic eosinophilia, and heterochromatic/pycnotic nuclei were found based on the histopathological damage score. After pinealectomy, we observed an increase in oxidative stress and a decrease in AChE levels in the brain.

Aim: Anabolic androgenic steroids (AAS) are synthetic derivatives of the male sex hormone testosterone. Androgens and anabolic steroids have been used for therapeutic purpose with few exceptions. However, the abuse of AAS is a remarkably prevalent problem, particularly among athletes and adolescents. Supraphysiological doses of AAS exert profound effects on mental state and behaviors such as depression, anxiety, aggressiveness, and cognitive deterioration.Objective: In the present investigation, we studied the impact of one of the AAS compounds, i.e., 17α-methyltestosterone on acetylcholinesterase (AChE) enzyme activity in different brain parts of mice, namely, forebrain, hippocampus, midbrain, and hindbrain.Methods: The adult female mice were assigned to four experimental groups to which different doses of 17α-MT (0.5, 5.0 and 7.5 mg/kg bwt, respectively) were administrated s.c. for 30 days. A significant increase in AChE activity in forebrain and midbrain (low and medium dose treatment) suggests a reduction of cholinergic neurotransmission efficiency due to decrease in acetylcholine levels in trans-synaptic cleft. Further, concurrent reduction in AChE activity was observed in whole brain, hippocampus, and hindbrain of 17α-MT-treated mice suggests the impairment in neuronal transmission. Since the regulation of cholinergic system through acetylcholine hydrolysis has been largely attributed to AChE activity, a significant reduction in its activity may lead to stress-related anxiety, memory loss with some cognitive and behavioral aspects in the mice.Conclusion: Based on the observed results, we propose that 17α-MT, an alkylated steroid compound, has a negative impact on AChE enzyme activity in different parts of mice brain, leading to impairment in neuronal transmission.

Many efforts have been made to evaluate organophosphorus (OP) toxicity by sensitive biomarkers. Therefore, the aim of this work is to evaluate cholinesterase and paraoxonase (PON1) enzymes activities as diagnostic tools in acute & chronic organophosphorus toxicity with poisoning severity assessment.The present study was conducted on 90 adult men after taking their informed consent, they were divided into three groups; group (I) included thirty patients who were acutely exposed to organophosphorus insecticides (OPI). Group (II) included thirty farm workers chronically exposed to OP. Group (III) included thirty healthy matched volunteers served as control group. The severity of symptoms and signs of acute OP poisoning was graded into mild, moderate and severe grade. Long term pesticide exposure intensity was estimated depending on mixing, application methods, repair activities and use of personal protective equipment (PPE). Exposure intensity score = (Mix + Apply + Repair) х PPE. Butyrylcholinesterase (BuChE), acetylcholinesterase (AChE) and paraoxonase (PON1) enzymes activities were determined. The result of the current study revealed significant decrease in BuChE, AChE and PON1 in groups I and II when compared to group III. Moreover, BuChE and AChE enzymes were significantly decreased in group I when compared to group II. Significant positive correlation was detected between AChE enzyme activity and both BuChE and PON1 enzymes activities in group I. However, group IIshowed significant positive correlations between BuChE enzyme activity and the activity of each of AChE and PON1. Moreover, there was significant decrease in AChE enzyme activity in severe cases compared to mild and moderate cases in group I. Nevertheless, group II registered significant decrease in BuChE, AChE and PON-1 when Pesticide Exposure Intensity Score is more than 10.

Earthworms are important and useful soil organisms, but in agricultural soils, they are potentially exposed to a wide variety of pesticides. Insecticides represent the highest threat to earthworms and many are neurotoxic. There is a need for a reliable, relevant, simple biomarker to assess the sub-lethal effects of neurotoxic insecticides on earthworms under laboratory or field conditions. The Aporrectodea caliginosa earthworms were exposed to 0 (control), 0.5×, 1× (normal field application rate), and 5× concentrations of a carbamate (Pirimor®) and an organophosphate (Lorsban®) insecticides. The nerve conduction velocity (NCV) of the medial giant fibers of A. caliginosa earthworm was recorded on days 0, 1, 2, 3, 4, and 7 to quantify sub-lethal neurotoxic effects. Acetylcholinesterase (AChE) enzyme activity of A. caliginosa homogenates was measured at the conclusion of the experiment. Pirimor® but not Lorsban® induced a significant decrease in NCV on days 3, 4, and 7 at 1× and 5× doses. A significant dose-dependent decrease was observed on AChE activity to Pirimor® at the doses used but not Lorsban®. A clear relationship is observed between AChE activity and NCV in the case of Pirimor®. This study showed that NCV is a sensitive biomarker that correlates well with classical biomarker measurements such as AChE enzyme activity. This technique could be used to study the impact of insecticides on earthworms and also their recovery.

Aim This study aims to comprehensively evaluate the effects of ferulic acid (FA) on acetylcholinesterase (AChE) enzyme activity and amyloid beta (Aβ) peptide plaque formation in an in vitro model of Alzheimer's disease (AD). Background ADis a progressive neurological condition marked by disrupted cholinergic signaling, accumulation of Aβ peptide, and tau protein hyperphosphorylation. Currently, no direct anti-Alzheimer drug that effectively prevents the cognitive decline from AD has been reported. To combat this, a multi-target drug addressing several molecular aspects would be ideal for AD. Natural compounds are preferred over synthetic drugs due to their accessibility, cost-efficiency, and lower toxicity The proven association between polyphenolconsumption and the prevention of AD has led to the investigation of the effect of FA, a polyphenolic compound, on acetylcholinesterase enzyme activity and Aβ peptide formation, the key targets of AD. Materials and method The free radical scavenging ability of FA was assessed by xanthine oxidase inhibitory activity. Furthermore, FA was also evaluated for its inhibitory activity against AChE enzyme and amyloid beta peptide formation to evaluate the neuroprotective potential of FA. Results The results showed that FA has the potential to be an AChE inhibitor, thus helping in blocking the activity of AChE and also reducing the incidence of amyloid beta plaque formation. Furthermore, the compound also exhibited a significant antioxidant property which was demonstrated by the xanthine oxidase enzyme inhibitory effect. Conclusion From the observed results, FA has significant antioxidant and neuroprotective effects which are compared with those of their respective standards. More research is required to determine the efficacy and safety of this compound as a treatment for neurodegenerative diseases like AD because the precise mechanism and degree of its AChE inhibitory effects in the brain are still elusive. A potent, selective, and effective drug is desperately needed to treat patients with AD and those at risk of developing the disease.

BackgroundCholinergic drugs are the most commonly used drugs for the treatment of Alzheimer’s disease (AD). Therefore, a better understanding of the cholinergic system and its relation to both AD-related biomarkers and cognitive functions is of high importance.ObjectivesTo evaluate the relationships of cerebrospinal fluid (CSF) cholinergic enzymes with markers of amyloidosis, neurodegeneration, neurofibrillary tangles, inflammation and performance on verbal episodic memory in a memory clinic cohort.MethodsIn this cross-sectional study, 46 cholinergic drug-free subjects (median age = 71, 54% female, median MMSE = 28) were recruited from an Icelandic memory clinic cohort targeting early stages of cognitive impairment. Enzyme activity of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) was measured in CSF as well as levels of amyloid-β1–42 (Aβ42), phosphorylated tau (P-tau), total-tau (T-tau), neurofilament light (NFL), YKL-40, S100 calcium-binding protein B (S100B), and glial fibrillary acidic protein (GFAP). Verbal episodic memory was assessed with the Rey Auditory Verbal Learning (RAVLT) and Story tests.ResultsNo significant relationships were found between CSF Aβ42 levels and AChE or BuChE activity (p > 0.05). In contrast, T-tau (r = 0.46, p = 0.001) and P-tau (r = 0.45, p = 0.002) levels correlated significantly with AChE activity. Although neurodegeneration markers T-tau and NFL did correlate with each other (r = 0.59, p < 0.001), NFL did not correlate with AChE (r = 0.25, p = 0.09) or BuChE (r = 0.27, p = 0.06). Inflammation markers S100B and YKL-40 both correlated significantly with AChE (S100B: r = 0.43, p = 0.003; YKL-40: r = 0.32, p = 0.03) and BuChE (S100B: r = 0.47, p < 0.001; YKL-40: r = 0.38, p = 0.009) activity. A weak correlation was detected between AChE activity and the composite score reflecting verbal episodic memory (r = −0.34, p = 0.02). LASSO regression analyses with a stability approach were performed for the selection of a set of measures best predicting cholinergic activity and verbal episodic memory score. S100B was the predictor with the highest model selection frequency for both AChE (68%) and BuChE (73%) activity. Age (91%) was the most reliable predictor for verbal episodic memory, with selection frequency of both cholinergic enzymes below 10%.ConclusionsResults indicate a relationship between higher activity of the ACh-degrading cholinergic enzymes with increased neurodegeneration, neurofibrillary tangles and inflammation in the stages of pre- and early symptomatic dementia, independent of CSF Aβ42 levels.

In our study, it was aimed to investigate the effects of cefazolin, cefuroxime, and cefoperazon injected to rats on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzyme activities in the heart, brain, eye, liver, and kidney tissues of rats. Liver AChE activity at the 1st and 3rd hours of cefuroxime groups was higher than the control group at the same time (p <.05). The AChE activity of the heart tissue decreased in the cefazolin group compared to the control group at the same hour, whereas it increased in the cefuroxime group (p <.05). AChE activities of kidney tissue of cefazolin and cefuroxime groups were lower than those of the same control group on the 3rd and started to increase on the next hours (p <.05). BChE activity is measured in tissues increased within the first three hours and decreased significantly within the first hour in the cefoperazone group (p <.05).

Background and Aims: Some pesticide chemical compounds, such as organophosphates and carbamates, interfere with or inhibit cholinesterase activity. Employees working in pesticide factories are one of the groups at risk of pesticide poisoning. This study aimed to evaluate the effect of work on exposure to toxins on the serum level of erythrocyte cholinesterase activity. Materials and Methods: This research was conducted on Mehriz Elixir Pesticide Factory workers. Blood samples were taken from 76 employees in 2 groups as a control group, and 38 workers were exposed to organophosphorus toxins three months after starting work in a factory. Cholinesterase activity was analyzed using the Elman method, and data were analyzed using SPSS software version 22. Results: The mean age of the subjects was 35.07 years, which was 35.26 in the exposure group (n = 38) and 34.89 in the control group (n = 38). The activity of acetylcholinesterase enzyme in the control group at 0, 10, and 20 minutes was 12.78 ku/l, 14.24 ku/l, and 15.45 ku/l, respectively. The acetylcholinesterase enzyme activity in the exposed group was 10.77 ku/l, 10.40 ku/l, and 10.36 ku/l at 0, 10, and 20 minutes, respectively. At all stages, the mean acetylcholinesterase activity in the control group was higher than the exposed group, but significant differences were observed at 10 and 20 minutes between the 2 groups. Conclusion: Exposure of workers to organophosphate inhibits acetylcholinesterase, which manifests by a decrease in the activity of this enzyme.

The purpose of our study is to assist in understanding the effects of wireless electromagnetic waves on carbonic anhydrase (CA) and acetylcholinesterase (AChE) enzymes activities in the different tissues of the rats. For this purpose, two different groups each of which contains eight rats (n=8) were formed as being control group and wireless electromagnetic wave-administered group. The rats were necropsied after 60min from the injection of chemicals into the rats intraperitoneally. The different tissues of the rats were extracted. CA and AChE enzymes activities were measured for each tissue. All the experimental results were provided in mean±S.D. Statistical significance was identified to be P<0.05. It was observed that there were significant changes of enzyme activities in wireless-administered group in salivary gland, stomach, colon, liver, and striated muscle tissues.

The aim of our studies is to assist in understanding the effects of wireless electromagnetic waves on activities of carbonic anhydrase (CA) and acetylcholinesterase (AChE) enzymes in the different tissues r elated to respiratory, excretory and reproductive systems of the wistar albino rats. For this respect, 2 different groups each of which contains 8 rats (n = 8) were formed as control group and wireless electromagnetic wave administered groups. The rats were necropsied and the different tissues of the rats were extracted. Activities of carbonic anhydrase and acetylcholinesterase enzymes were measured for each tissue by using the hydratase, esterase and acetylcholinioidide methods. All the experimental results were provided in mean ± standard deviation ( ± SD). Statistical significance was identified to be p<0.05. It was observed that there were significant decreases of enzyme activities in wireless administered group in brain, spleen, kidney, and trachea tissues according to CA and AChE levels.

Biochemical stress response in freshwater fish Channa punctatus induced by aqueous extracts of Euphorbia tirucalli plant

The aim of this study was to investigate the effects of oxytocin (OT), atosiban, which is an OT receptor antagonist, and OT-atosiban chemicals injected to rats on the activities of carbonic anhydrase (CA) and acetylcholinesterase (AChE) enzymes in liver and kidney tissues of rats. For this purpose, four different groups, each consisting of six rats (n=6), were formed (control group, OT administered group, atosiban administered group, and both OT and atosiban administered group). The rats were necropsied 60min after intraperitoneal injection of chemicals into the rats. Liver tissues of rats were extracted. CA and AChE enzyme activities were measured for each tissue by using hydratase, esterase, and acetylcholiniodide methods. Activity values for each enzyme obtained were statistically calculated.

Organophosphates (OPs) and carbamates (CBs) are known to reduce acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) and increase Glutathione S-Transferase (GST) activities in the exposed individuals. We have evaluated AChE, BChE and GST activities in long-term pesticide exposed tea garden workers of sub-Himalayan West Bengal and tried to relate enzyme activities with polymorphisms in GST M1 and GST T1 genes. Enzyme activities were measured spectro-photometrically in blood samples of sprayers and pluckers in relation to age-matched control individuals. Allele-specific multiplex-PCR was employed to score the presence of GST M1 and GST T1 alleles. Significantly lower (p < 0.01) level of AChE and BChE activities were found in the worker group (3.15 ± 0.69 µmol/min/mL of lysate and 1.2 ± 0.54 µmol/min/mL of plasma respectively) compared to the control group (3.74±0.26 µmol/min/mL of lysate and 2.21 ±0.27 µmol/min/mL of plasma respectively). Whereas, significantly higher (p < 0.01) level of GST activity was detected in the worker group (3.83 ±0.97 lU/gm of Hb) comprising of 61 individuals compared to control group (2.47 ±0.29 lU/gm of Hb) comprising of equal number of individuals. Out of total 122 individuals, 34.43% had both GST M1 and GST T1, 23.77% had single-null genotype for GST T1, 27.05% had the single-null genotype for GST M1 and 14.75% individuals had double-null genotypes. We found no significant difference in enzyme activities among the four genotypes when test and control groups were compared. This study for the first time explored changes in AChE, BChE and GST activities in tea garden workers exposed chronically to OPs and CBs in the studied region. Our study indicated that exposure to OP and CB pesticides leads to changes in AChE, BChE and GST activities and the presence or absence of GST M1 and GST T1 alleles are possibly not related to the changes in AChE, BChE and GST activities.

Aims: To investigate the effect of the methanolic extracts of Caryota no (CN) seeds negative geotaxis, fecundity and acetylcholinesterase (AChE) using Drosophila melanogaster (DM). Study Design: Experimental design.&#x0D; Place and Duration: Sample: African Centre of Excellence for Phytomedicine Research and Development, University of Jos, Jos Plateau State Nigeria between June 2018 and February 2019 Methodology: 50 flies were exposed in each vial to the following concentrations: 300 mg, 350 mg, 400 mg, 500 mg and 600 mg of methanolic extracts in 5 replicates for 7 days with daily recording of mortality. Total protein assays were carried out by Randox method from the supernatant from homogenized whole flies. In vivo antioxidant activity study was conducted by measuring level of acetylcholinesterase (AChE) enzyme activity from supernatants of whole fly homogenates using a spectrophotometer at specific wavelengths over a 2 minute duration. The values were derived as part of the total protein value. Negative geotaxis was done by the climbing assay and fecundity was examined by rate of emergence of larva after exposure of the flies to treatment. The statistical difference among test groups was presumed at P &lt; .05.&#x0D; Results: The methanolic extract of CN caused nonsignificant (P = .33) decrease in total protein levels compared to the control. There were also nonsignificant decreases in AChE (P = .30) activity, negative geotactic (P = .80) behaviour and nonsignificant increase in fecundity (P = .17) in the methanolic extract-treated flies compared to the controls.&#x0D; Conclusion: It can therefore be concluded that the methanolic extract of Caryota no nonsignificantly improved fertitity and reduced negative geotaxis and AChE activity in DM.

Immobilization stress-induced changes in brain acetylcholinesterase activity and cognitive function in mice