Hydrolysis Of Acetylcholine Research Articles

Acetylcholine Hydrolysis and Antidote Delivery Using a Histidine-Resorcinarene-based Nanocontainer

A new nanocarrier was developed for the delivery of an antidote for poisoning with organophosphorus compounds. The nanocarrier acts as an artificial esterase, hydrolyzing acetylcholine and releasing the antidote (atropine) when acetylcholine level is high. The nanocarrier was made using histidine-containing resorcinarene as a building block. Histidine-resorcinarene was preorganized in a microemulsion media and then polymerized with phenylboronic acid, which acts as a linker between the resorcinarene molecules. Antidote (atropine) was incorporated into the nanocarrier with an encapsulation efficiency of 52.2%. At a neutral pH of 7.4, the nanocarrier hydrolyzes acetylcholine to choline and acetic acid. The acid triggers dissociation of the boronate bonds to dissociate, resulting in nanocarrier degradation and the release of 64.4% of the antidote. Transmission electron microscopy (TEM), dynamic and static light scattering (DLS and SLS, respectively), and IR spectroscopy were utilized to characterize the structure of the nanocarriers. Cyclic voltammetry and NMR spectroscopy were employed to evaluate its ability to hydrolyze acetylcholine. Through fluorescence and NMR spectroscopy, it was demonstrated that the nanocarrier could release substrates (fluorescein and atropine) in in response to the presence of acetylcholine.

Preliminary exploration of the expression of acetylcholinesterase in normal human T lymphocytes and leukemic Jurkat T cells.

The classic enzymatic function of acetylcholinesterase (AChE) is the hydrolysis of acetylcholine (ACh) in the neuronal synapse. However, AChE is also present in nonneuronal cells such as lymphocytes. Various studies have proposed the participation of AChE in the development of cancer. The ACHE gene produces three mRNAs (T, H and R). AChE-T encodes amphiphilic monomers, dimers, tetramers (G1 A, G2 A and G4 A) and hydrophilic tetramers (G4 H). AChE-H encodes amphiphilic monomers and dimers (G1 A and G2 A). AChE-R encodes a hydrophilic monomer (G1 H). The present study considered the differences in the mRNA expression (T, H and R) and protein levels of AChE, as well as the molecular forms of AChE, the glycosylation pattern and the enzymatic activity of AChE present in normal T lymphocytes and leukemic Jurkat E6-1 cells. The results revealed that AChE enzymatic activity was higher in normal T lymphocytes than in Jurkat cells. Normal T cells expressed AChE-H transcripts, whereas Jurkat cells expressed AChE-H and AChE-T. The molecular forms identified in normal T cells were G2 A (5.2 S) and G1 A (3.5 S), whereas those in Jurkat cells were G2 A (5.2 S), G1 A (3.5 S) and G4 H (10.6S). AChE in Jurkat cells showed altered posttranslational maturation since a decrease in the incorporation of galactose and sialic acid into its structure was observed. In conclusion, the content and composition of AChE were altered in Jurkat cells compared with those in normal T lymphocytes. The present study opened new avenues for exploring the development of novel therapeutic strategies against T-cell leukemia and for identifying potential molecular targets for the early detection of this type of cancer.

Detection of Hydrogen Ions Resulting from the Inhibition of Acetylcholinesterase Using Indium Tin Oxide-Coated Nanostructures for Pesticide Sensing

The neurotransmitter acetylcholine (ACh) is hydrolyzed by acetylcholinesterase (AChE) into acetic acid and choline to terminate synaptic transmission (1). Organophosphorus (OPs) found in pesticides induce toxicity in organisms by irreversibly binding to AChE, leading to the phosphorylation of the serine residue at the active site. ACh cannot undergo hydrolysis by the phosphorylated enzyme, resulting in excessive stimulation of nerves and muscles, akin to a nerve agent (2). Our objective is to develop a rapid detection tool for determining the levels of pesticide residues in dairy products. In this study, we measured the release of hydrogen ions (H+) resulting from the hydrolysis of ACh by AChE, serving as an indirect method to evaluate the inhibitory effect of OPs on AChE. To detect the signals of H+ released during ACh decomposition, we used ITO-coated nanostructures modified with 3-aminopropyl trimethoxysilane (APTMS) as a sensing substrate integrated into the extended gate of a field-effect transistor (EGFET) biosensor. The amino group at the end of APTMS can bond with H+. By monitoring the changes in electrical signal from the EGFET biosensor, it is possible to know the concentration of H+ in the solution. The output characteristics (Id-Vd) of the APTMS-modified ITO-VASiNW EGFET were measured using ACh solutions at different concentrations. A correlation was observed between Id and ACh concentration within the tested range, exhibiting a sensitivity of 0.016 mA1/2 / mM and an R2 value of 0.9. A lower concentration of H+ indicates a more pronounced inhibitory effect on AChE by OPs and suggests higher levels of pesticide residues. References (1) Colović, M. B., Krstić, D. Z., Lazarević-Pašti, T. D., Bondžić, A. M., & Vasić, V. M. (2013). Acetylcholinesterase inhibitors: pharmacology and toxicology. Curr Neuropharmacol, 11(3), 315-335.(2) Rajagopalan, V., Venkataraman, S., Rajendran, D. S., Vinoth Kumar, V., Kumar, V. V., & Rangasamy, G. (2023). Acetylcholinesterase biosensors for electrochemical detection of neurotoxic pesticides and acetylcholine neurotransmitter: A literature review. Environmental Research, 227, 115724. Figure 1

Atomistic Origins of Resurrection of Aged Acetylcholinesterase by Quinone Methide Precursors.

Nerve agents are organophosphates (OPs) that act as potent inhibitors of acetylcholinesterase (AChE), the enzyme responsible for the hydrolysis of acetylcholine. After inhibition, a dealkylation reaction of the phosphorylated serine, known as the aging of AChE, can occur. When aged, reactivators of OP-inhibited AChE are no longer effective. Therefore, the realkylation of aged AChE may offer a pathway to reverse AChE aging. In this study, molecular modeling was conducted to propose new ligands as realkylators of aged AChE. We applied a methodology involving docking and quantum mechanics/molecular mechanics (QM/MM) calculations to evaluate the resurrection kinetic constants and ligand interactions with OP-aged AChE, comparing them to data found in the literature. The results obtained confirm that this method is suitable for predicting kinetic and thermodynamic parameters of ligands, which can be useful in the design and selection of new and more effective ligands for AChE realkylation.

Naturally Occurring Cholinesterase Inhibitors from Plants, Fungi, Algae, and Animals: A Review of the Most Effective Inhibitors Reported in 2012-2022.

Since the development of the "cholinergic hypothesis" as an important therapeutic approach in the treatment of Alzheimer's disease (AD), the scientific community has made a remarkable effort to discover new and effective molecules with the ability to inhibit the enzyme acetylcholinesterase (AChE). The natural function of this enzyme is to catalyze the hydrolysis of the neurotransmitter acetylcholine in the brain. Thus, its inhibition increases the levels of this neurochemical and improves the cholinergic functions in patients with AD alleviating the symptoms of this neurological disorder. In recent years, attention has also been focused on the role of another enzyme, butyrylcholinesterase (BChE), mainly in the advanced stages of AD, transforming this enzyme into another target of interest in the search for new anticholinesterase agents. Over the past decades, Nature has proven to be a rich source of bioactive compounds relevant to the discovery of new molecules with potential applications in AD therapy. Bioprospecting of new cholinesterase inhibitors among natural products has led to the discovery of an important number of new AChE and BChE inhibitors that became potential lead compounds for the development of anti-AD drugs. This review summarizes a total of 260 active compounds from 142 studies which correspond to the most relevant (IC50 ≤ 15 μM) research work published during 2012-2022 on plant-derived anticholinesterase compounds, as well as several potent inhibitors obtained from other sources like fungi, algae, and animals.

Design, synthesis and anti-Alzheimer's disease activity evaluation of C-3 arylated huperzine A derivatives

A series of C-3 arylated huperzine A (HPA) derivatives (1−30) were designed and synthesized in good yields via palladium-catalyzed Suzuki cross-coupling reaction. Cholinesterase inhibitory and neuroprotective activities of all 30 derivatives were evaluated. Cholinesterase inhibition results revealed that derivatives 2 and 15 exhibited dual inhibitory activity against both acetylcholinesterase (AChE inhibition: 2, IC50 = 1.205 ± 0.395 μM; 15, IC50 = 0.225 ± 0.062 μM) and butyrylcholinesterase (BChE inhibition: 2, IC50 = 8.598 ± 3.605 μM; 15, IC50 = 4.013 ± 0.068 μM), a feature not observed in huperzine A. Molecular docking results indicated that the introduction of aryl groups enhanced the affinity of the derivatives for the acyl-binding pocket of BChE, thereby limiting the hydrolysis of acetyl choline. However, these derivatives exhibited poor performance in cytotoxicity and neuroprotection assays.

Low-dose aspirin increases olfactory sensitivity in association with enhanced neurogenesis and reduced activity of AChE in the experimental aging mice

This study examined the effect of aspirin on olfactory discrimination and neurogenesis in experimental aging mice. The results from the buried food test (BFT) and odour preference test (OPT) indicated that aspirin treatment enhanced the olfactory functions in the experimental animals. The immunohistochemical assessment revealed increased amounts of doublecortin (DCX)-positive immature neurons and bromodeoxyuridine (BrdU)/neuronal nuclei (NeuN) double-positive new neurons in the olfactory bulb (OB) of aspirin-treated animals compared to the control group. Besides, aspirin treatment resulted in a decreased number of ionized calcium-binding adaptor molecule (Iba)-1 positive microglia, a main cellular element of neuroinflammation in the brain compared to the control group. In addition, the increased activities of key antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in OB of the aspirin-treated mice were evident. Acetylcholinesterase (AChE) is a key enzyme that hydrolyses acetylcholine (ACh), an important neurotransmitter, involved in cognition, and olfaction. While the enhanced activity of AChE has been linked to the severity of dementia and OB defects, the biochemical assessments indicated reduced activity of AChE in OB-derived homogenates of the aspirin-treated experimental animals than that of the control group. Taken together, this study validates the proneurogenic, cholinergic, and anti-neuroinflammatory properties of aspirin in the OB which can be considered in boosting neural regeneration and management of olfactory deficits in aging and various disorders.

Multi Targeted Ligands for Potential Inhibition of Dipeptidyl Peptidase 4, Acetylcholinesterase and Cyclooxygenase 2

Two series of novel compounds were designed by combining indomethacin and ibuprofen with sixteen sulfa drugs. These compounds were systematically evaluated through target fishing using the Pharm Mapper, leading to the identification of DPP-4, AChE, and COX-2 as potential targets. Molecular docking was performed to evaluate the binding affinity of designed compounds against the identified three target proteins. The results revealed that the designed compounds exhibited binding affinities ranging from ~8 to -12kcal/mol, 12 to 13 kcal/mol and 8 to 11kcal/mol for DPP-4, AChE and COX-2 respectively. The binding affinities were found to be comparable or higher than binding affinity of co-crystallized ligand, which was found to be ~10, 12 and 9 kcal/mol respectively. Further investigation into the binding modes of these compounds was carried out. Notably, for DPP-4 complexes, interactions with Arg125, Glu205, and Glu206 were observed which are essential for substrate and inhibitor binding. For AChE complexes, interactions involved crucial His447 residues, essential for acetylcholine hydrolysis. In the case of COX-2, hydrogen bond interaction was noted with Arg120 located at the entrance of the hydrophobic channel. Despite favorable binding potentials, ADME profiling highlighted five compounds (1A, 1F, 1G, 1H, and 1O) with drug-like characteristics but lacking blood-brain barrier permeation ability. Out of five compounds, 1H stood out, demonstrating superior binding affinity and interactions vital residues necessary for catalytic activity of three enzymes. Thus, 1H emerges as a promising candidate for Multi-Targeted Drug-Like (MTDL) development aimed at addressing diabetes mellitus related dementia.

Bifunctional enzyme-mimicking metal-organic frameworks for sensitive acetylcholine analysis

The development of nanomaterials with multi-enzyme-like activity is crucial for addressing challenges in multi-enzyme-based biosensing systems, including cross-talk between different enzymes and the complexities and costs associated with detection. In this study, Pt nanoparticles (Pt NPs) were successfully supported on a Zr-based metal-organic framework (MOF-808) to create a composite catalyst named MOF-808/Pt NPs. This composite catalyst effectively mimics the functions of acetylcholinesterase (AChE) and peroxidase (POD). Leveraging this capability, we replaced AChE and POD with MOF-808/Pt NPs and constructed a biosensor for sensitive detection of acetylcholine (ACh). The MOF-808/Pt NPs catalyze the hydrolysis of ACh, resulting in the production of acetic acid. The subsequent reduction in pH value further enhances the POD-like activity of the MOFs, enabling signal amplification through the oxidation of a colorimetric substrate. This biosensor capitalizes on pH variations during the reaction to modulate the different enzyme-like activities of the MOFs, simplifying the detection process and eliminating cross-talk between different enzymes. The developed biosensor holds great promise for clinical diagnostic analysis and offers significant application value in the field.

PED: a novel predictor-encoder-decoder model for Alzheimer drug molecular generation.

Alzheimer's disease (AD) is a gradually advancing neurodegenerative disorder characterized by a concealed onset. Acetylcholinesterase (AChE) is an efficient hydrolase that catalyzes the hydrolysis of acetylcholine (ACh), which regulates the concentration of ACh at synapses and then terminates ACh-mediated neurotransmission. There are inhibitors to inhibit the activity of AChE currently, but its side effects are inevitable. In various application fields where Al have gained prominence, neural network-based models for molecular design have recently emerged and demonstrate encouraging outcomes. However, in the conditional molecular generation task, most of the current generation models need additional optimization algorithms to generate molecules with intended properties which make molecular generation inefficient. Consequently, we introduce a cognitive-conditional molecular design model, termed PED, which leverages the variational auto-encoder. Its primary function is to adeptly produce a molecular library tailored for specific properties. From this library, we can then identify molecules that inhibit AChE activity without adverse effects. These molecules serve as lead compounds, hastening AD treatment and concurrently enhancing the AI's cognitive abilities. In this study, we aim to fine-tune a VAE model pre-trained on the ZINC database using active compounds of AChE collected from Binding DB. Different from other molecular generation models, the PED can simultaneously perform both property prediction and molecule generation, consequently, it can generate molecules with intended properties without additional optimization process. Experiments of evaluation show that proposed model performs better than other methods benchmarked on the same data sets. The results indicated that the model learns a good representation of potential chemical space, it can well generate molecules with intended properties. Extensive experiments on benchmark datasets confirmed PED's efficiency and efficacy. Furthermore, we also verified the binding ability of molecules to AChE through molecular docking. The results showed that our molecular generation system for AD shows excellent cognitive capacities, the molecules within the molecular library could bind well to AChE and inhibit its activity, thus preventing the hydrolysis of ACh.

New paradigms in the study of the cholinergic system and metabolic diseases: Acetyl-and-butyrylcholinesterase.

Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are enzymes that belong to the neuromuscular cholinergic system, their main function is to hydrolyze the neurotransmitter acetylcholine (ACh), through their hydrolysis these enzymes regulate the neuronal and neuromuscular cholinergic system. They have recently attracted considerable attention due to the discovery of new enzymatic and nonenzymatic functions. These discoveries have aroused the interest of numerous scientists, consolidating the relevance of this group of enzymes. Recent investigations have revealed a positive correlation between several risk factors for metabolic syndrome (MetS) and the expression of cholinesterases (ChE's), which underscore the impact of high ChE's activity on the pro-inflammatory state associated with MetS. In addition, the excessive hydrolysis of ACh and other choline esters (succinylcholine, propionylcholine, butyrylcholine, etc.) by both ChE's results in the overproduction of fatty acid precursor metabolites, which facilitate the synthesis of very low-density lipoproteins and triacylglycerols. Participation in these processes may represent the link between ChE's and metabolic disorders. However, further scientific research is required to fully elucidate the involvement of ChE's in metabolic diseases. This review aims to collect recent research studies that contribute to understanding the association between the cholinergic system and metabolic diseases.

Transport mechanism of presynaptic high-affinity choline uptake by CHT1.

Choline is a vital nutrient and a precursor for the biosynthesis of essential metabolites, including acetylcholine (ACh), that play a central role in fetal development, especially in the brain. In cholinergic neurons, the high-affinity choline transporter (CHT1) provides an extraordinarily efficient reuptake mechanism to reutilize choline derived from intrasynaptical ACh hydrolysis and maintain ACh synthesis in the presynapse. Here, we determined structures of human CHT1 in three discrete states: the outward-facing state bound with the competitive inhibitor hemicholinium-3 (HC-3); the inward-facing occluded state bound with the substrate choline; and the inward-facing apo open state. Our structures and functional characterizations elucidate how the inhibitor and substrate are recognized. Moreover, our findings shed light on conformational changes when transitioning from an outward-facing to an inward-facing state and establish a framework for understanding the transport cycle, which relies on the stabilization of the outward-facing state by a short intracellular helix, IH1.

THE Point prevalence of acetylcholinesterase inhibition, neuropathy and safety awareness among flower farm workers in naivasha, nakuru county, kenya.

Acetyl cholinesterase catalyses the hydrolysis of acetylcholine in the nerve synapses, thereby terminating nerve impulse, however, it is inhibited by organophosphates and carbamates. This study aimed to assess the level of acetyl cholinesterase inhibition and the resultant neuropathy in flower farm workers as well as the farmers’ awareness of safety measures and predisposing factors while handling pesticides. A cross-sectional study was conducted involving 217 participants from different flower farms. Structured questionnaire was used to collect data to assess level of safety and predisposing awareness and to assess for neuropathy. Blood samples were collected to determine the cholinesterase levels using spectrophotometry technique at 405 nm. The mean serum cholinesterase level in flower farm workers in Naivasha was 5873.26 U/L. There was a positive correlation (R= 0.07) between the numbers of years worked in the industry and serum cholinesterase levels as shown by Karl Pearson`s coefficient. The point prevalence of cholinesterase inhibition in Naivasha was 14% indicating a significant risk of adverse health effects. The level of safety awareness and predisposing factors among workers on pesticide exposure was 65%, suggesting that while the majority of workers had some level of safety awareness, there is still room for improvement. The point prevalence of neuropathy was 38% of the participants presenting with varying symptoms of adverse effects of pesticide exposure. Musculoskeletal impairment was leading at 19.81%, followed by skin irritation at 13.36%.These findings highlighted the need for better safety measures and awareness campaigns in the flower farm industry to reduce the risk of acetyl cholinesterase inhibition.

Identification of a novel drug molecule for neurodegenerative disease from marine algae through in-silico analysis

Cognitive functions are lost due to the rapid hydrolysis of acetylcholine including Acetylcholinesterase (AChE) and Butyrylcholinesterase (BChE). Marine algae-derived compounds were reported for their neuroprotective activities and hence they can be utilised for treating neurodegenerative ailments like Alzheimer’s Disease and Parkinson’s Disease which are due to the loss of cognitive functions. Major attention is currently paid to seaweeds due to their health benefits and high nutritional values. Sea weeds are of a rich sense of natural bioactive compounds which antioxidants, pharmaceutical compounds, flavonoids and alkaloids. They also contain a high amount of vitamins A, D, E, C and Ca, K, Mg and Fe. Regular consumption of a marine algae-based diet may boost immunities. In searching for natural cholinesterase inhibitors, the present study is focussed on some marine bioactive compounds reported from brown, red and green algae. Molecular docking studies have been carried out along with molecular dynamics simulations studies and binding energy calculations resulting in three best bioactive compounds when AChE is used as the target. The results are compared with cocrystal studies. Two best compounds, namely, Diphlorethohydroxycarmalol and Phlorofucofuroeckol from the brown seaweeds are identified as the potential lead compounds for neurodegenerative diseases, Alzheimer’s and Parkinson’s. Communicated by Ramaswamy H. Sarma

Eucalyptus globulus leaf-isolated isorhapontin serves as a natural insecticide via acetylcholinesterase inhibition

Acetylcholinesterase (AChE) inhibitors cause insect death by preventing the hydrolysis of the neurotransmitter acetylcholine, which overstimulates the nervous system. In this study, isorhapontin, isolated from E. globulus leaves, was evaluated as a natural insecticide with AChE inhibition at 12.5 μM. Using kinetic analyses, we found that isorhapontin acted as a competitive inhibitor that binds to the active site of AChE. The inhibition constant (Ki) was 6.1 μM. Furthermore, isorhapontin and resveratrol, which have basic skeletons, were predicted to bind to the active site of AChE via molecular docking. A comparison of the hydrogen bonding between the two stilbenes revealed characteristic differences in their interactions with amino acids. In isorhapontin, Trp83, Gly149, Tyr162, Tyr324, and Tyr370 interacted with the sugar moiety. These results suggest that with further development, isorhapontin can be used as an insecticide alternative.

Electrochemical Acetylcholinesterase Sensors for Anti-Alzheimer's Disease Drug Determination.

Neurodegenerative diseases and Alzheimer's disease (AD), as one of the most common causes of dementia, result in progressive losses of cholinergic neurons and a reduction in the presynaptic markers of the cholinergic system. These consequences can be compensated by the inhibition of acetylcholinesterase (AChE) followed by a decrease in the rate of acetylcholine hydrolysis. For this reason, anticholinesterase drugs with reversible inhibition effects are applied for the administration of neurodegenerative diseases. Their overdosage, variation in efficiency and recommendation of an individual daily dose require simple and reliable measurement devices capable of the assessment of the drug concentration in biological fluids and medications. In this review, the performance of electrochemical biosensors utilizing immobilized cholinesterases is considered to show their advantages and drawbacks in the determination of anticholinesterase drugs. In addition, common drugs applied in treating neurodegenerative diseases are briefly characterized. The immobilization of enzymes, nature of the signal recorded and its dependence on the transducer modification are considered and the analytical characteristics of appropriate biosensors are summarized for donepezil, huperzine A, rivastigmine, eserine and galantamine as common anti-dementia drugs. Finally, the prospects for the application of AChE-based biosensors in clinical practice are discussed.

In-Vitro Acetylcholine Esterase Enzyme Inhibition Potential of Siddha Formulation Vilvaver Chooranam: A Neuroprotective Assay

Background and Aim: Alzheimer's disease (AD) is a progressive neurodegenerative condition evidenced by significant cognitive dysfunction. The state of cognitive impairment is made worse by increased levels of the enzyme acetylcholinesterase (AChE), which is crucial in the hydrolysis of the neurotransmitter acetylcholine (ACh). Siddha therapy gaining higher momentum in recent days due to its global acceptance considering its broad spectral safety and therapeutic window. Siddha originated from the southern geographic landscape of Asia now spreading its wings across the bounders in managing dreadful diseases like AD. The main objective of the present study is to evaluate AChE inhibition of the Siddha formulation Vilvaver Chooranam (VVC). Materials & Methods: In-Vitro Acetylcholine esterase enzyme inhibition Potential of Siddha formulation Vilvaver Chooranam by Ellman’s method. Results:Results obtained from the study clearly demonstrate that the formulation VVC has shown promising acetylcholinesterase at stipulated concentration dose-dependently. Maximum percentage inhibition of about 54.53 ± 3.475 % was observed at 500μg/ml with the IC50 value of 411.9 ± 30.6 µg/ml when compared to that of the Physostigmine, a known AChE Inhibitor with a maximum inhibition 93.44 ± 4.434 % at the concentration of 40μg/ml with the IC50 value of 10.38± 5.29 µg/ml. Conclusion: These findings demonstrate the remarkable potential of these extracts as valuable sources of antioxidants with interesting acetylcholinesterase inhibitory activity.

Novel ligand conjugated poly(propylene imine) dendrimers for brain targeted delivery of tacrine hydrochloride

Alzheimer's disease (AD) is the most common type of neurodegenerative disorder characterized by extracellular deposition of Amyloid β and intracellular deposition of neurofibrillary tangles. One of the most common etiological factors is rapid and excessive hydrolysis of acetylcholine by acetylcholinesterase (AChE) enzyme. Tacrine is one of the most potent and first USFDA approved centrally acting AChE inhibitor recommended for treatment of AD, but it is no longer in use as it causes severe peripheral side effects, mainly hepatotoxicity. In this research work, novel dendrimeric conjugates were developed with an aim of enhancing the brain targeting efficiency of tacrine and eliminating the peripheral side effects. Poly(propyleneimine) G5.0 dendrimers were conjugated with oleic acid as a brain targeting ligand, and the conjugation was confirmed by Fourier-transform infrared spectroscopy, 1H nuclear magnetic resonance spectroscopy and dynamic light scattering studies. Tacrine hydrochloride was loaded into the plain dendrimer as well as oleic acid conjugated dendrimer by physical encapsulation method. The in vitro AChE inhibitory study showed retention of the AChE inhibition ability of tacrine in dendrimeric conjugates. The safety of the drug loaded conjugates was confirmed by haemolytic, nasal ciliotoxicity and cytotoxicity studies. Further, cellular uptake studies in SH-SY5Y cells showed abundance of green cytoplasmic fluorescence, suggesting enhanced internalization of dendrimeric conjugates. The results of this research work support the further exploration of this novel dendrimers based conjugated delivery system for in vivo studies as a potential alternative to treat AD.

A MnO2 nanosheet-mediated CRISPR/Cas12a system for the detection of organophosphorus pesticides in environmental water.

Nowadays, easy, convenient, and sensitive sensing strategies are still critical for organophosphorus pesticides in environmental water samples. Herein, a novel organophosphorus pesticide (OP) assay based on acetylcholinesterase (AChE) and a MnO2 nanosheet-mediated CRISPR/Cas12a reaction is reported. The single-strand DNA (ssDNA) activator of CRISPR/Cas12a was simply adsorbed on the MnO2 nanosheets as the nanoswitches of the assay. In the absence of target OPs, AChE hydrolyzed acetylcholine (ATCh) to thiocholine (TCh), which reduced the MnO2 nanosheets to Mn2+, resulting in the release of the activator followed by activation of the CRISPR/Cas12a system. The activated Cas12a thereafter nonspecifically cleaved the FAM/BHQ1-labeled ssDNA (FQ-reporter), producing a fluorescence signal. Upon the addition of target OPs, the hydrolysis of ATCh by AChE was inhibited owing to OPs combining with AChE, and thus effective quantification of OPs could be achieved by measuring the fluorescence changes of the system. As a proof of concept, dichlorvos (DDVP) was chosen as a model OP analyte to address the feasibility of the proposed method. Attributed to the excellent trans-cleavage activity of Cas12a, the fluorescent biosensor exhibits a satisfactory limit of detection (LOD) for DDVP at 0.135 ng mL-1. In addition, the excellent recoveries for the detection of DDVP in environmental water samples demonstrate the applicability of the proposed assay in real sample research.

A molecular container providing supramolecular protection against acetylcholine hydrolysis.

Alzheimer's disease (AD) is characterized by cognitive decline, often attributed to the deficiency of acetylcholine, which can undergo hydrolysis by acetylcholinesterase (AChE) within the biological milieu. Here, we report a supramolecular strategy that takes advantage of confinement effects to inhibit such a hydrolysis process, shedding some light on AD therapy. A water-soluble and bowl-shaped molecule, hexacarboxylated tribenzotriquinacene (TBTQ-C6), was employed to shield acetylcholine (G1) from enzymatic degradation through host-guest binding interactions. Our study revealed highly efficient host-guest interactions with a binding ratio of 1 : 3, resulting in a significant reduction in acetylcholine hydrolysis from 91.1% to 7.4% in the presence of AChE under otherwise identical conditions. Furthermore, TBTQ-C6 showed potential for attenuating the degradation of butyrylcholine (G2) by butyrylcholinesterase (BChE). The broader implications of this study extend to the potential use of molecular containers in various biochemical and pharmacological applications, opening new avenues for research in the field of neurodegenerative diseases.