Acetylcholinesterase Enzyme Activity Research Articles

Pyrethroid-based insecticides exert transgenerational, persistent, and chronic effects in Caenorhabditis elegans

The use of agrichemical pyrethroid-based insecticides to combat crop infestations and ectoparasites in animals is increasing. In this context, two pyrethroid-based insecticides are widely used, λ-cyhalothrin and Cypermethrin. The mechanism of action of these insecticides is characterized by the opening of ion channels and death by neural hyperexcitability. In this study, we evaluated the toxicological effect of two pyrethroid-based insecticides λ-cyhalothrin and Cypermethrin in C. elegans aiming to evaluate the transgenerational (TG), neonatal (NN) and lifespan (LS) effects of these compounds. At the end of each exposure period, were evaluated behavioral biomarkers of body bends, pharyngeal pumping, and feeding behavior. Furthermore, the fluorescent expression of antioxidant enzymes (superoxide dismutase, catalase, and glutathione-S-transferase) and the fluorescent expression of PolyQ40 aggregates were quantified. Lastly, the activity of the enzyme acetylcholinesterase (AChE) was quantified. Changes in TG were more related to changes in AChE enzyme activity that probably were transferred to the offspring, altering behavioral biomarkers in the adult life of offspring from exposed parents. However, alterations in LS were related to the modulation of ion channels in a chronic way, exerting behavioral effects. In addition, both compounds increased the expression of PolyQ40 muscle aggregates in mutant worms. These proteins are related to the increased probability of the senile incidence of Huntington's Disease in genetically predisposed patients.

Role of cholinergic anti-inflammatory pathway in the regulation of vascular endothelial permeability and inflammation in type 2 diabetes

Abstract Despite tremendous advancement in glycemic control, anti-diabetic medications have failed to revert vascular impairment once triggered by the metabolic disorder. The cholinergic anti-inflammatory pathway (CAP) is a neurophysiological mechanism that regulates the immune system. Studies show that the CAP inhibits inflammation by suppressing cytokine synthesis via release of acetylcholine in organs of the reticuloendothelial system, including the heart, lungs and gastrointestinal tract. Upon release, acetylcholine interacts with α7 nicotinic acetylcholine receptors (α7nAChR), down-regulate pro-inflammatory cytokine synthesis and prevent tissue damage and inflammation. Although a CAP modulated the response to severe inflammation during sepsis, the role of this pathway in the regulation of type-2 diabetes (T2D)-induced increase in endothelial permeability is completely unknown. Our data showed that human coronary artery endothelial cells (HCAECs) from T2D displayed increased endothelial permeability and enhanced pro-inflammatory signaling in response to pro-coagulant thrombin and endotoxin lipopolysaccharide (LPS), respectively. We also found increased expression and activity of acetylcholinesterase enzyme, and no change in expression of α7nAChR in T2D endothelial cells (ECs) compared to ECs from normal subjects. Our research findings showed that the T2D increased the expression and activity of AChE enzyme in HCAECs, and thereby decreasing the availability of ACh at α7nAChR. In summary, our work offered novel insights and therapeutic targets that we think will lead to the development of more effective treatments with improved outcomes. Grant from CTRE Chicago State University, Chicago, IL

A chemiluminescence biosensor based on the peroxidase-like property of molybdenum disulfide/zirconium metal-organic framework nanocomposite for diazinon monitoring

BackgroundOrganophosphorus pesticides are widely used in agriculture owing to their high effectiveness as insecticides. Among these, diazinon is a common environmental contaminant that acts as an acetylcholinesterase (AChE) enzyme inhibitor. As the current methods are too expensive and time-consuming for routine analysis of diazinon, its trace monitoring by rapid and sensitive methods is critical to protect the environment and human health. ResultsA biosensor was introduced for the indirect detection of diazinon using a molybdenum disulfide/zirconium metal-organic framework (MoS2@MIP-202(Zr)) nanocomposite. The probe is based on the peroxidase mimic of the prepared nanocomposite on NaHCO3–H2O2 chemiluminescence system as well as the inhibitory effect of diazinon on the enzymatic activity of AChE. The chemiluminescence signal is gradually decreased with an increase in diazinon concentration, and there is a linear relationship between the analytical signal and diazinon concentration. Under the optimum conditions, the calibration plot is linear in the concentration range of 0.5–300.0 nmol L−1. The limit of detection and quantification limit of the method are 0.12 and 0.40 nmol L−1, respectively. The inter-day and intra-day relative standard deviations (% RSD n = 5, diazinon concentration; 100 nmol L−1) are 3.66 and 1.35%, respectively. The method was used for diazinon detection in real water samples, and the high relative recovery values for the spiked samples along with satisfactory results of a certified reference material analysis confirmed that the method is accurate and free from the matrix effect. Significance and noveltyA nano-probe based on the peroxidase-like property of MoS2@MIP-202(Zr) nanocomposite was developed for the first time for indirect detection of residue levels of diazinon in water samples. The high stability of the nanocomposite makes it a good alternative for natural peroxidase enzymes such as horseradish peroxidase with low stability.

Caffeic acid attenuates neuroinflammation and cognitive impairment in streptozotocin-induced diabetic rats: Pivotal role of the cholinergic and purinergic signaling pathways

The present study evaluated the effect of caffeic acid (CA) on behavioral learning and memory tasks in the diabetic state. We also evaluated the effect of this phenolic acid on the enzymatic activities of acetylcholinesterase, ecto-nucleoside triphosphate diphosphohydrolase, ecto-5-nucleotidase and adenosine deaminase as well as on the density of M1R, α7nAChR, P2×7R, A1R, A2AR, and inflammatory parameters in the cortex and hippocampus of diabetic rats. Diabetes was induced by a single intraperitoneal dose of streptozotocin (55 mg/kg). The animals were divided into six groups: control/vehicle; control/CA 10 and 50 mg/kg; diabetic/vehicle; diabetic/CA 10 and 50 mg/kg, treated by gavage. The results showed that CA improved learning and memory deficits in diabetic rats. Also, CA reversed the increase in acetylcholinesterase and adenosine deaminase activities and reduced ATP and ADP hydrolysis. Moreover, CA increased the density of M1R, α7nAChR, and A1R receptors and reversed the increase in P2×7R and A2AR density in both evaluated structures. In addition, CA treatment attenuated the increase in NLRP3, caspase 1, and interleukin 1β density in the diabetic state; moreover, it increased the density of interleukin-10 in the diabetic/CA 10 mg/kg group. The results indicated that CA treatment positively modified the activities of cholinergic and purinergic enzymes and the density of receptors, and improved the inflammatory parameters of diabetic animals. Thus, the outcomes suggest that this phenolic acid could improve the cognitive deficit linked to cholinergic and purinergic signaling in the diabetic state.

Minocycline alleviated scopolamine-induced amnesia by regulating antioxidant and cholinergic function

Background and aimMinocycline, a tetracycline derivative, has been found to exert neuroprotective properties. The current project aimed to assess the antioxidant status and cholinergic function in the amnesia induced by scopolamine. MethodsWe evaluated the passive avoidance performance, acetylcholine esterase (AChE) enzyme activity, and the oxidative stress indicators in the following groups: Normal control, scopolamine, and the treatment groups (the animals were given minocycline (10–30 mg/kg)). ResultsScopolamine (intraperitoneal) injection was associated with impairment of passive avoidance performance and neurotoxicity. Minocycline pronouncedly ameliorated scopolamine injury as presented by the increased latency time to darkness and stay time in lightness along with the decreased darkness entry. Moreover, minocycline decreased lipid peroxidation, while it elevated the levels of superoxide dismutase, AChE enzymes, and thiol groups in both the cortex and hippocampus. ConclusionOur data suggested that minocycline modulated the antioxidant status and AChE in the brains, which may contribute to its protective effects against scopolamine-induced amnesia.

Enzyme colorimetric cellulose membrane bioactivity strips based on acetylcholinesterase immobilization for inhibitors preliminary screening

To quickly screen the active pharmaceutical ingredient that can be used as acetylcholinesterase inhibitors (AChEIs) to treat Alzheimer's disease, an enzyme colorimetric cellulose membrane bioactivity strip (CBS) was developed for simple and rapid screening of AChEIs. The amino group of acetylcholinesterase (AChE) undergoes Schiff base reaction with the aldehyde group on the oxidized cellulose membranes, then the AChE was covalently cross-linking on the surface of cellulose membranes, enabling the screening based on Ellman's enzyme colorimetric method. When the enzyme activity of AChE was inhibited after incubation with inhibitors, the hydrolysis of S-Acetylthiocholine iodide decreased, consequently, the 5-thio-2-nitrobenzoic acid generated by the reaction with 5,5′-dithiobis (2-nitrobenzoic acid) also decreased, leading to a decreased color intensity. In addition, CBSs had fast chromogenic time, excellent specificity, and extraordinary storage stability. Tacrine and Donepezil were used as representative inhibitors during the detection, while their IC50 and limit of detection were determined.Therefore, our work not only established a platform for effective preliminary screening of AChEIs but also inspired the further development of other cellulose membrane-based biosensors.

Organochlorine pesticides induce thyroid tumors through oxidative stress; an in vivo and in silico study.

Thyroid disease is one of the most common endocrine problems around the world. Among the numerous factors, exposure to environmental elements such as pesticides is associated with an increase in the incidence of thyroid disorders. The aim of the present study was to investigate the role of organochlorine pesticides (OCPs) in induction of oxidative stress (OS) and development of thyroid tumors. This case-control study was conducted on 61 patients with papillary thyroid carcinoma (PTC), 70 patients with benign thyroid nodules (BTN), and 73 healthy individuals as control. Seven derived OCPs residues measured by gas chromatography (GC), and enzyme activities of acetylcholinesterase (AChE), superoxide dismutase3 (SOD3), catalase (CAT), glutathione peroxidase3 (GPx3) and paraoxonase1 (PON1) and also, non-enzymatic antioxidant including; malondialdehyde (MDA), total antioxidant capacity (TAC), protein carbonyl (PC), and nitric oxide (NO) biomarkers in all participants were investigated. Furthermore, all of the above enzymes were docked against measured OCPs. The results revealed that β-HCH, γ-HCH, 2,4 DDE, 4,4 DDE, 2,4-DDT, and 4,4-DDT levels along with MDA, NO, and PC levels were elevated, while AChE, SOD3, GPx3, CAT, and PON1 activities and TAC levels were decreased in the PTC and BTN groups compared with the control group. Therefore, OCPs might play a role in the development of thyroid tumors through several mechanisms including generation of OS. Importantly, in silico analysis confirmed the in vivo findings.

Green Synthetized Selenium Nanoparticles Using Syzygium aromaticum (Clove) Extract Reduce Pentylenetetrazol-Induced Epilepsy and Associated Cortical Damage in Rats

We aimed to investigate the potential anticonvulsant effect of green synthetized selenium nanoparticles (SeNPs) using Syzygium aromaticum extract (SAE) (SAE-SeNPs) against epileptic seizures and cortical damage induced by pentylenetetrazole (PTZ) injection in rats and its mechanism. A total of 84 rats were divided into six groups; control, PTZ-exposed group, SAE + PTZ-treated group, sodium selenite (Na2SeO3) + PTZ-treated group, SAE-SeNPs + PTZ-treated group, and diazepam + PTZ-treated group. SAE-SeNPs significantly increase (p < 0.05) the latency time to seizures and reduce both the seizure duration and death rate, which were enhanced by the PTZ injection. SAE-SeNPs counteracted the PTZ-induced changes in the oxidants and antioxidants. Furthermore, SAE-SeNPs significantly restored (p < 0.05) the pro-inflammatory cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor-α) to their normal levels and suppressed the activity of the glial fibrillary acidic protein showing their inhibitory effect on the epilepsy-associated inflammation. In addition, SAE-SeNPs significantly reduced (p < 0.05) PTZ-induced cortical cell apoptosis, as revealed by a reduction in the pro-apoptotic Bax and caspase-3 levels, and an elevation of the anti-apoptotic Bcl-2 level. Moreover, SAE-SeNPs significantly modulate (p < 0.05) the PTZ-induced changes in the neurotransmitter norepinephrine level and acetylcholinesterase enzymatic activity. These data concluded the anticonvulsant activity of SAE-SeNPs via their antioxidant, anti-inflammatory, and anti-apoptotic effects, along with their ability to modulate neurotransmitters.

Presenilin 1 Modulates Acetylcholinesterase Trafficking and Maturation.

In Alzheimer's disease (AD), the reduction in acetylcholinesterase (AChE) enzymatic activity is not paralleled with changes in its protein levels, suggesting the presence of a considerable enzymatically inactive pool in the brain. In the present study, we validated previous findings, and, since inactive forms could result from post-translational modifications, we analyzed the glycosylation of AChE by lectin binding in brain samples from sporadic and familial AD (sAD and fAD). Most of the enzymatically active AChE was bound to lectins Canavalia ensiformis (Con A) and Lens culinaris agglutinin (LCA) that recognize terminal mannoses, whereas Western blot assays showed a very low percentage of AChE protein being recognized by the lectin. This indicates that active and inactive forms of AChE vary in their glycosylation pattern, particularly in the presence of terminal mannoses in active ones. Moreover, sAD subjects showed reduced binding to terminal mannoses compared to non-demented controls, while, for fAD patients that carry mutations in the PSEN1 gene, the binding was higher. The role of presenilin-1 (PS1) in modulating AChE glycosylation was then studied in a cellular model that overexpresses PS1 (CHO-PS1). In CHO-PS1 cells, binding to LCA indicates that AChE displays more terminal mannoses in oligosaccharides with a fucosylated core. Immunocytochemical assays also demonstrated increased presence of AChE in the trans-Golgi. Moreover, AChE enzymatic activity was higher in plasmatic membrane of CHO-PS1 cells. Thus, our results indicate that PS1 modulates trafficking and maturation of AChE in Golgi regions favoring the presence of active forms in the membrane.

Effect of Rosa damascena Extract on Rat Model Alzheimer's Disease: A Histopathological, Behavioral, Enzyme Activities, and Oxidative Stress Study.

The purpose of the current study is to investigate the effect of aquatic Rosa damascena extract against the oxidative damage induced by aluminum chloride intoxication in Alzheimer's model of Wister rats. Rats were divided randomly into seven groups (n = 10). Control group received no treatment, sham group received distilled water orally, aluminum group (AL) was administered AlCl3 (100 mg/kg) orally, extract 1 and 2 groups were treated with only aqueous R. damascena extract (DRE) (500 and 1000 mg/kg), and treatment 1 and 2 groups received aqueous R. damascena extract (500 and 1000 mg/kg) and AlCl3 (100 mg/kg) orally. The brain tissues were sampled for histopathological examination, and biochemical analysis was conducted for estimating the enzyme activities of acetylcholinesterase and catalase (CAT), the levels of GSH and MDA, and ferric reducing antioxidant power. According to the results of behavioral tests, AL administration showed a reduction in spatial memory and remarkably increased the time needed for reaching the invisible platform. The administration of Al-induced oxidative stress and an increase of the enzyme activity of AChE. Al administration increased AChE level from 1.176 ± 0.173 to 3.62 ± 0.348, which was a significant rise. However, treating with the extract at the dose of 1000 mg/kg downregulated it to 1.56 ± 0.303. Administration of the R. damascene extract caused an increased level of catalase and glutathione levels in treatment groups, attenuated MDA level, and regulated AChE activity. Our results illustrate that administration of R. damascene extract has a protective effect against the oxidative damage induced by AlCl3 intoxication in Alzheimer's model.

A novel RP-HPLC method for quantification of cholinesterase activity in human blood: An application for assessing organophosphate and carbamate insecticide exposure.

Several methods have been reported to estimate Acetylcholinesterase (AChE) enzyme activity in blood samples. The Ellman assay is the most important among all but with several shortcomings, and there is a need to develop a method which is accurate, sensitive and quick for analyzing AChE. Therefore, we have developed an assay utilizing RP-HPLC with UV detection for the determination of AChE activity. This method measured the conversion of 1-naphthol acetate to 1-naphthol to estimate AChE activity in blood samples. Performance was judged on the basis of reproducibility, sensitivity, accuracy, and the ability to screen enzyme activity within 20minutes. A series of experiments were performed, varying the concentration of blood and substrate, with optimal sensitivity using 50 μM substrate and 10μL blood. The validation parameters such as linearity (R2 of ≥ 0.9842 for 1-naphthol and ≥ 0.9897 for 1-naphthol acetate), precision (94.21-96.41%), accuracy (85.2%-99.6% and 82.6%-99.3% for 1-naphthol and 1-naphthol acetate respectively), and robustness were validated according to International Conference on Harmonization (ICH) guidelines. Blood samples were collected from healthy people, farmers exposed to spraying of pesticides, and suicidal patients who ingested pesticides and were hospitalized and were analyzed by the developed method. The AChE level was approximately 21 units/mL compared to 24units/mL in controls, whereas suicidal patients showed the least AChE levels of 1 unit/mL. The employment of this method is recommended for estimating AChE level on various matrices.

Protective effects of cinnamon on acetylcholinesterase activity and memory dysfunction in diabetic rats.

Regarding neurocognitive and immunomodulatory properties of cinnamon (Cinn) we aimed to investigate whether cinnamon regulates acetylcholinesterase (AChE) activity, and oxidative abnormalities with concomitant memory dysfunction in streptozotocin (STZ)-induced diabetes. Forty-seven male adult rats were divided into seven groups (n=8 animals): Control group: in these non-diabetic rats only saline 0.9% NaCl was gavaged, Diabetic (Dia) group: diabetic rats in them saline 0.9% NaCl was gavaged for six weeks. Dia-Cinn 100, Dia-Cinn 200, and Dia-Cinn 400, Dia-Met groups: in these diabetic rats the extract (100, 200, 400mg/kg respectively) or metformin (300mg/kg) was gavaged for six weeks. Passive avoidance performance, AChE enzyme activity, and oxidative indicators were examined among the groups. Vs. the control group, blood glucose level and stay time in the dark were remarkably increased in Dia group whereas the latency time was decreased. Meanwhile, antioxidant levels (superoxide dismutase, catalase, and thiols) noticeably decreased in the Dia group compared to the Control group. On the other hand, Cinn extract espicailly at the highest dose recovered the changes similar to those found in the metformin-treated group. These findings proposed that the cinnamon hydro-ethanolic extract promotes memory recovery in diabetic conditions through the atteuation of the AChE activity and oxidative injury.

PRUNETIN ATTENUATES D-GALACTOSE-INDUCED BRAIN AGING VIA INHIBITING AMYLOID-Β AND TAU PROTEIN AGGREGATION

The objective of this investigation was to assess the effect of prunetin in D-galactose-induced brain aging in rats and its regulating mechanisms. D-galactose (200 mg kg-1 body wt.) was given orally daily for 45 days to accelerate aging, and prunetin (10, 20,40, and 80 µg kg-1 body wt. respectively) was administered orally. The anti-oxidant and anti-brain aging activities of prunetin in serum were measured by the estimation of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and malondialdehyde (MDA) levels and brain tissues were measured by western blot analysis and histopathological studies. Prunetin therapy decreases elevated levels of glucose, C-reactive protein, cholesterol, and triglyceride levels in the D-galactose-induced rats. D-galactose suppresses the levels of superoxide dismutase, glutathione peroxidase, catalase and total antioxidant capacity in rats; these levels were elevated by treating with prunetin. Malondialdehyde levels were elevated in D-galactose-induced rats. Prunetin significantly decreases the malondialdehyde levels in rat brain tissue. Prunetin decreases mitochondrial dysfunction induced by D-galactose, by improving the activities of Na+K+-ATPase and acetylcholinesterase enzyme activity. Western blot analysis results showed that the degree of brain tissue damage was significantly reduced by prunetin. The results of our study indicated that prunetin treatment reduced oxidative stress by exerting a protective effect against D-galactose-induced aging in rats, by significantly decreasing amyloid - β and tau protein levels in the brain tissue. Prunetin exhibits anti-oxidant activity by increasing anti-oxidant enzymes.

Piranhea trifoliata extracts ameliorate muscular decline in Drosophila melanogaster exposed to Paraquat.

In this study, we have demonstrated, for the first time, the muscular protective effects of Piranhea trifoliata bark extract against Paraquat (PQ)-induced oxidative stress in Drosophila melanogaster. Exposure of D. melanogaster (Canton Special) to PQ caused oxidative stress, as evidenced by protein carbonyl and elevated acetylcholinesterase (AChE) activity levels. However, a diet supplemented with the P. trifoliata extracts (0.1 mg/ml) for 10 days ameliorates protein carbonyl levels and enzymatic activities of AChE and citrate synthase to prevent PQ damage. Also, P. trifoliata bark extracts showed in phytochemical assays the presence of phenols, at 46.06 mg EAG/g extract of total phenolic compounds, and a 40% 2,2-diphenyl-1-picryl-hydrazyl scavenging effect. The study showed the muscular protective function of the P. trifoliata extracts in D. melanogaster exposed to PQ. On the basis of the results, we contemplate that the bark of P. trifoliata might prevent and ameliorate human diseases caused by oxidative stress. The muscular action of the P. trifoliata extract can be attributed to the antioxidant constituents, while the precise mechanism of its action needs further investigation.

EFFECT OF SUB-LETHAL CONCENTRATION OF NICOTINE IN SURFACE WATERS ON ENZYMATICAL CHANGES IN TISSUES OF Labeo rohita FINGERLINGS

The current study aims to analyse the impact of the sub-lethal concentration of nicotine in surface waters on enzymatical changes in tissues of Labeo rohita fingerlings. The experimental fish Labeo rohita fingerlings were exposed to sub-lethal nicotine concentrations in surface water (0.93 mgL-1) for 60 days. Tissues (liver, gill and kidney) were harvested at different intervals (15, 30, 45 and 60 days of nicotine exposure) to analyze metabolic enzymes, TBARS and antioxidant enzymes. Further acetylcholinesterase enzyme activity was examined in the brain tissue. The metabolic enzymes such as AST and ALT activities increased significantly (p<0.05). In contrast, ACP and ALP activities declined in nicotine-exposed fish of liver, gill and kidney at every interval than in control fish. The peroxidation marker TBARS activity was significantly reduced, meanwhile the activities of enzymatic antioxidants such as SOD, CAT and GPx and non-enzymatic antioxidant GSH activities declined considerably in sub-lethal concentration nicotine-exposed fish tissues (liver, gill and kidney) when compared to control fish. Further, the activity of AChE also declined in the brain of nicotine-exposed fish than in control fish. The results indicate that exposure to the sub-lethal concentration of nicotine in surface water can cause an imbalance in the activities of metabolic enzymes, TBARS, antioxidant enzymes and AChE in fish.

INSECTICIDAL PROPERTY OF TERPENES AGAINST MAIZE WEEVIL, SITOPHILUS ZEAMAIS (MOTSCHULSKY)

Synthetic pesticides are used indiscriminately in insect pest management, which damages the ozone layer and causes resistance in target organisms as well as neurotoxicity, carcinogenicity, teratogenicity and mutagenesis in non-target organisms. Due to these negative consequences, research is now focused on using plant-based techniques to control insect pests. The maize weevil, Sitophilus zeamais (Coleoptera: Curculionidae), was tested in the laboratory to determine the insecticidal effects of two pure essential oil constituents, namely α-pinene and β-caryophyllene. These two terpenes were tested against S. zeamais for their toxic, ovipositional, developmental, and feeding inhibitory effects. When S. zeamais adults were fumigated for 24 and 48hrs, the median lethal concentrations (LC50) of α-pinene and β-caryophyllene were 0.412 and 0.305 μlcm-3 and 0.486 and 0.315 μlcm-3 air respectively. When S. zeamais adults were exposed for 24 and 48hrs in a contact toxicity assay, the LC50 values for α-pinene and β-caryophyllene were 0.388 and 0.256 μlcm-2 and 0.308 and 0.216 μlcm-2 area respectively. Adults exposed to sub-lethal concentrations of both terpenes experienced decreased acetylcholine esterase (AChE) enzyme activity. In S. zeamais, α-pinene and β-caryophyllene decreased oviposition, progeny output and eating. According to this study, α-pinene and β-caryophyllene can be used to make environmentally acceptable formulations and as a substitute for synthetic insecticides.

Hippeastrum stapfianum (Kraenzl.) R.S.Oliveira & Dutilh (Amaryllidaceae) Ethanol Extract Activity on Acetylcholinesterase and PPAR-α/γ Receptors.

Hippeastrum stapfianum (Kraenzl.) R.S.Oliveira & Dutilh (Amaryllidaceae) is an endemic plant species from the Brazilian savannah with biological and pharmacological potential. This study evaluated the effects of ethanol extract from H. stapfianum leaves on acetylcholinesterase enzyme activity and the action on nuclear receptors PPAR-α and PPAR-γ. A gene reporter assay was performed to assess the PPAR agonist or antagonist activity with a non-toxic dose of H. stapfianum ethanol extract. The antioxidant capacity was investigated using DPPH• scavenging and fosfomolybdenium reduction assays. The identification of H. stapfianum's chemical composition was performed by gas chromatography-mass spectrometry (GC-MS) and HPLC. The ethanol extract of H. stapfianum activated PPAR-α and PPAR-γ selectively, inhibited the acetylcholinesterase enzyme, and presented antioxidant activity in an in vitro assay. The major compounds identified were lycorine, 7-demethoxy-9-O-methylhostasine, and rutin. Therefore, H. stapfianum is a potential source of drugs for Alzheimer's disease due to its ability to activate PPAR receptors, acetylcholinesterase inhibition activity, and antioxidant attributes.

Effect of Chlorpyrifos Insecticide On The Ossification of Wader Pari Fish Vertebrae (Rasbora Lateristriata Bleeker, 1854)

Due to high growth, chlorpyrifos was one of the solution to increase food production. However, this compound is dangerous because it belongs to organophosphate group which blocking the activity of acetylcholinesterase (ache) enzyme and caused nervouse failure to animal. Due to heavy rain this insecticide will run-off and contaminate the body of the river and effect the aquatic live. Wader pari was non-target animal of chlorpyrifos. The purpose of this study was to determine the effect of chlorpyrifos exposure on the behaviour of wader's feeding and swimming response, as well as the morphology and ossification process of the vertebrae. The study was conducted for 7 days.  4 groups of fish were exposed to chlorpyrifos with concentrations of 0, 0.001, 0.005 and 0.01 ppm. Observation of the ossification and morphological process of vertebrae is carried out using AR-Ab bone staining. Observation of wader’s behaviours is carried out using descriptive analysis. It was found that the fish which exposed to chlorpyrifos showed some behavioural abnormalities in swimming and feeding response. Fish exposed to 0.005 ppm chlorpyrifos showed vertebral abnormalities which is haemal kyphosis. Kyphosis refers to Λ-shaped vertebral curvature Fragkoulis et.al, (2019). However, the ossification process that represented by Ar-Ab staining are going well.Â

Nanoparticle-induced enhancement of cholinesterase activity in the presence of malathion: A potential nerve agent therapeutic

Organophosphate nerve agents are associated with assassination, terrorism and chemical warfare, but there has been slow progress in developing a broad-spectrum response to poisoning. For some nerve agents the oxime component of the therapy may not be effective, limiting the effectiveness of emergency treatment that is desperately needed. An alternative therapy may be possible based on accelerating enzyme (acetylcholinesterase) catalysis in unaffected adjacent enzymes. Herein we demonstrate a restoration of acetylcholinesterase activity in malathion-inhibited cell membrane preparations by the administration of functional nanoparticles. The molecularly imprinted polymer nanoparticles were designed to bind selectively to designated enzyme epitopes. Enzyme activity of membrane-bound acetylcholinesterase was measured in the presence of the organophosphate malathion and the selected nanoparticles. Enzymatic acceleration of the cholinesterase was observed at 162 ± 17 % the rate of erythrocyte ghosts without bound nanoparticles. This may restore sufficient acetylcholine hydrolysis to mitigate the effects of poisoning, offsetting the acetylcholine accumulation resulting from enzyme inhibition.

Role of non-neuronal cholinergic system in the early stage response of epithelial-mesenchymal transformation related markers in A549 cells induced by coal particles

ObjectiveThis study was aimed to investigate the role of non-neuronal cholinergic system (NNCS) in the early stage response of epithelial-mesenchymal transformation (EMT) related markers in human lung adenocarcinoma A549 cells induced by coal particles. MethodsA549 cells were exposed to different concentrations of GBW11110K, GBW11126D and exogenous acetylcholinesterase (AChE) (the exposure doses were determined according to the results of CCK-8 experiment, and the doses that had no significant effects on cell viability were selected) for 24 h. After exposure, the indexes of oxidative stress (SOD and MDA), inflammatory factors (IL-6 and TNF-α), EMT marker proteins (E-cadherin and vimentin), AChE enzymatic activity and mRNA expression levels of different types of acetylcholine receptors (CHRM3, CHRM5, CHRNA5, CHRNA7, CHRNA9 and CHRNB2) were determined. ResultsGBW11110K and GBW11126D exposure could lead to the following injury effects: the levels of oxidative stress and inflammatory factors changed to a certain extent (SOD decreased gradually, while MDA, IL-6 and TNF-α increased). The protein level of E-cadherin decreased while the vimentin level increased (P < 0.05), suggesting the occurrence of EMT. The AChE enzymatic activity decreased gradually. The expression of acetylcholine receptor mRNA changed as follows (GBW11110K/GBW11126D: CHRM3 (↑↑), CHRM5 (↓↓), CHRNA5 (↓↓), CHRNA7 (↓↓), CHRNA9 (– ↑), CHRNB2 (– –). The addition of exogenous AChE recombinant protein could antagonize the damage effects caused by the coal particles to a certain extent. ConclusionThe coal particle exposure could induce the change of oxidative stress response, inflammatory response and EMT related markers, down-regulate the AChE enzymatic activity, and interfere the mRNA expression levels of AChRs in A549 cells. The addition of exogenous AChE recombinant protein could reverse the above effects to a certain extent.