Cholinergic Enzymes Research Articles

Phenolic constituents and inhibitory effects of the leaf of Rauvolfia vomitoria Afzel on free radicals, cholinergic and monoaminergic enzymes in rat's brain in vitro.

Rauvolfia vomitoria is a medicinal plant used traditionally in Africa in the management of several human diseases including psychosis. However, there is inadequate scientific information on the potency of the phenolic constituents of R.vomitoria leaf in the management of neurodegeneration. Therefore, this study characterized the phenolic constituents and investigated the effects of aqueous and methanolic extracts of R.vomitoria leaf on free radicals, Fe2+-induced lipid peroxidation, and critical enzymes linked to neurodegeneration in rat's brain in vitro. The polyphenols were evaluated by characterizing phenolic constituents using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD). The antioxidant properties were assessed through the extracts ability to reduce Fe3+ to Fe2+; inhibit ABTS, DPPH, and OH radicals and Fe2+-induced lipid peroxidation. The effects of the extracts on AChE and MAO were also evaluated. The phenolic characterization of R.vomitoria leaf revealed that there were more flavonoids present. Both aqueous and methanolic extracts of R.vomitoria leaf had inhibitory effects with the methanolic extract having higher significant (p≤0.05) free radicals scavenging ability coupled with inhibition of monoamine oxidases. However, there was no significant (p≤0.05) difference obtained in the inhibition of lipid peroxidation and cholinesterases. This study suggests that the rich phenolic constituents of R.vomitoria leaf might contribute to the observed antioxidative and neuroprotective effects. The methanolic extract was more potent than the aqueous extract; therefore, extraction of R.vomitoria leaf with methanol could offer better health-promoting effects in neurodegenerative condition.

Mixture of Organophosphates Chronic Exposure and Pancreatic Dysregulations in Two Different Population Samples.

Organophosphates (OP) are a major agrochemical. The application of OP pesticides is expected to increase multifold in the coming decades. The etiology of diabetic diseases is attributed to multiple factors including OP pesticide exposure. The present study investigates pancreatic dysregulation with respect to exocrine enzymes and diabesity in groups of Pakistani and Cameroonian people exposed to a mixture of OP pesticides. Nine hundred and four OP exposed individuals were enrolled for this cross-sectional study after due consent and approval from an ethical review committee. Pesticides' residues were measured by GC-MS spectrometry. Cholinergic enzymes were measured by Elman's method. Serum glucose, insulin, serum amylase, lipase, and triglyceride were measured by spectrophotometry and ELISA; HOMA-IR was determined in OP exposed and non-exposed participants. Stata 15 and R 3.2.0 software were used for statistical analysis of the data. Malathion, chlorpyrifos, and parathion residues were evident in plasma samples. RBC-acetylcholinesterase was significantly depressed in OP exposed groups. In both population samples, investigated pancreatic functions were found to be statistically significantly more dysregulated than non-exposed. OP exposure indicated risk of diabetes and insulin, glycaemia, adiponectin, triglycerides, and TNF-α dysregulations. The study concludes that both OP exposed population groups exhibited a mixture of OP residues and pancreatic dysregulation, although the effect was more pronounced in the Cameroonian population. In addition, serum lipase has a positive correlation with OP exposure and diabetes and may be suggested as an alternate/additional diagnostic marker for diabesity under OP exposure. However, screening of other environmental co-factors with OP for pancreatic dysregulation is suggested.

Effect of aqueous extracts of Pleurotus ostreatus and Lentinus subnudus on activity of adenosine deaminase, arginase, cholinergic enzyme, and angiotensin-1-converting enzyme.

Pleurotus ostreatus (P. ostreatus) and Lentinus subnudus (L. subnudus) commonly consumed as food or as food supplement have been reported in folklore for their antihypertensive property with limited scientific proof. This study aims to unravel the antihypertensive mechanism of P. ostreatus and L. subnudus in vitro. The antioxidant properties of P. ostreatus and L. subnudus were established via standard antioxidant assays. Also, the effect of P. ostreatus and L. subnudus extracts on relevant enzymes associated to the development of hypertension were evaluated. Findings from this study revealed that P. ostreatus and L. subnudus extracts exhibited antihypertensive and antioxidant properties. Meanwhile, according to our results, various bioactive compounds present in P. ostreatus and L. subnudus could be responsible for the observed in vitro antihypertensive property. PRACTICAL APPLICATIONS: P. ostreatus and L. subnudus are the most commonly consumed mushrooms by the rural dwellers in South Western Nigeria, perhaps, based on their nutritive value and health-enhancing benefits. This paper showed that P. ostreatus and L. subnudus possess antihypertensive and antioxidant properties. Thus, their consumption as foods or food supplements may provide therapeutic benefits for hypertensive patients. Therefore, P. ostreatus and L. subnudus are promising candidates for the development of nutraceuticals.

Microwave-assisted synthesis, biological assessment, and molecular modeling of aza-heterocycles: Potential inhibitory capacity of cholinergic enzymes to Alzheimer's disease

A highly regioselective solvent-free microwave-assisted synthesis of pyrazoles and tetrahydroindazoles based on the condensation of 1,3-diketones with arylhydrazines is described. Compounds were evaluated as cholinesterase inhibitors in order to identify an alternative treatment for Alzheimer's disease. All compounds displayed moderated acetylcholinesterase inhibitory activity and most of the compounds displayed remarkable butyrylcholinesterase inhibitory activity and selectivity. The compounds 3y and 3i with IC50 of 1.65 and 3.59 µM, respectively, were the most active and selective compounds as butyrylcholinesterase inhibitors. Likewise, the compounds were tested as antioxidants agents, results showed that they have the ability to trap free-radicals. Molecular Docking studies showed a key π-π stacking interaction of most of the compounds with residue Trp82 within of butyrylcholinesterase active site. Molecular quantum similarity field, global and local reactivity descriptors, and the Fukui functions were calculated in the Density Functional Theory framework to analyze the reactivity patterns along with the molecular set.

Analysis of PON1 gene polymorphisms (rs662 and rs854560) and inflammatory markers in organophosphate pesticides exposed cohorts from two distinct populations.

Background and objectivesOrganophosphate (OPs) anticholinesterases are one of the main groups of pesticides used in agriculture. Harmful effects of OPs on health have been attributed primarily for irreversible inhibition of acetylcholinesterase (AChE) at nerve synapse. However, studies have shown that inhibition of AChE alone cannot explain all the maladies encountered in prolonged exposure to OPs. Predisposition to population heterogeneity and irregularities in various biochemicals like paraoxonases and inflammatory biochemicals are the possible affects of OPs long term exposure that may lead to sequels of diseases and are less addressed in literature. The study was aimed to assess the cholinergic enzymes (AChE and BChE), PON1, and inflammatory markers (IL1β, IL6, TNFα, CRP, Apo AI, Apo B) and determine the toxicogenetics association of PON1 gene (rs 662 and rs 85456) to chronically OPs exposed groups from Pakistan and Cameroon. Materials and methodsAChE, BChE and PON1 were measured by colorimetric method using spectrophotometry. Inflammatory markers were determined by Elisa assay. PCR-restriction fragment length polymorphism (PCR-RFLP) using salting out method was employed for SNP genotyping. ResultsThe results revealed the significant (p ≤ 0.05) inhibition of cholinergic enzymes PON 1 was found to be 6.91 ng/mL±1.03 and 2.84 ng/mL±1.40 (mean ± SD) in Pakistan and Cameroon groups respectively. IL6, TNFα, CRP were increased and Apo AI was less while Apo B was increased in OP exposed groups in both population groups. SNPs analysis of PON1 showed significant differences in allelic and genotype frequencies of OPs exposed and non-exposed groups. ConclusionsPON1 was noticeably less in Cameroonian than Pakistani, albeit both groups have significant decrease in PON1 actity. In addition, the study concludes that OPs induce low grade inflammation, an aetiology of many diseases. Selected PON1 SNPs analysis showed a significant toxicogenetics association with OPs exposure marker enzymes. The results of this study may help in regulation of usage of OPs anticholinesterases in different populations. The study will further open new avenues in toxicogenetic and exploration of SNPs based strategies on organophosphate intoxication.

Assessment of cholinesterase activity and hepatic biofunction in dogs naturally infected with Babesia gibsoni

Canine babesiosis is a common tick-borne disease affecting canidae of tropical and subtropical regions of the world. This study investigated the activity of cholinergic system enzymes and hepatic biofunction in dogs naturally infected with Babesia gibsoni. An infected group comprised 10 male dogs (stray and shepherd), and 10 healthy dogs as control were included in the study. Hematological parameters and activities of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) were measured. The biomarkers of hepatic function including the activities of various enzymes, total bilirubin levels, and albumin contents were also assessed. The infected animals revealed a significant decrease in RBC count, packed cell volume, hemoglobin concentration, albumin content, and BchE activity compared with the control animals. However, the activities of AChE and hepatic enzymes as well as bilirubin levels were significantly higher in the diseased animals. Our results indicate that B. gibsoni infection can change the activities of AChE and BChE and damage liver. Further studies are required to elucidate the role of the cholinergic system in the pathogenesis of canine babesiosis.

Catechol protects against iron-mediated oxidative brain injury by restoring antioxidative metabolic pathways; and modulation of purinergic and cholinergic enzymes activities.

This study was aimed at investigating neuroprotective effect of catechol on redox imbalance, cholinergic dysfunctions, nucleotide hydrolysing enzymes activities, and dysregulated metabolic pathways in iron-mediated oxidative brain injury. Oxidative injury was induced in brain tissues by incubating with 0.1mm FeSO4 and treated with different concentrations of catechol. Catechol significantly elevated glutathione level, superoxide dismutase and catalase activities, while depleting malondialdehyde and nitric oxide levels. It also inhibited the activities of acetylcholinesterase, butyrylcholinesterase, and ATPase, with concomitant elevation of ENTPDase activity. GC-MS analysis revealed that treatment with catechol completely depleted oxidative-generated lipid metabolites. While LC-MS analysis revealed depletion of oxidative-generated metabolites in brain tissues treated with catechol, with concomitant restoration of oxidative-depleted metabolites. Catechol also led to reactivation of oxidative-inactivated taurine and hypotaurine, purine, glutathione, glycerophospholipid, nicotinate and nicotinamide, fructose and mannose, pyrimidine metabolisms and pentose phosphate pathways. Catechol was predicted in silico to be permeable across the blood-brain barrier with a predicted oral LD50 value of 100mg/kg and a toxicity class of 3. These results suggest the neuroprotective effects of catechol in iron-mediated oxidative brain injury.

Oxidative Stress and Analysis of Selected SNPs of ACHE (rs 2571598), BCHE (rs 3495), CAT (rs 7943316), SIRT1 (rs 10823108), GSTP1 (rs 1695), and Gene GSTM1, GSTT1 in Chronic Organophosphates Exposed Groups from Cameroon and Pakistan.

The detrimental effects of organophosphates (OPs) on human health are thought to be of systemic, i.e., irreversible inhibition of acetylcholinesterase (AChE) at nerve synapses. However, several studies have shown that AChE inhibition alone cannot explain all the toxicological manifestations in prolonged exposure to OPs. The present study aimed to assess the status of antioxidants malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) (reduced), catalase, and ferric reducing antioxidant power (FRAP) in chronic OP-exposed groups from Cameroon and Pakistan. Molecular analysis of genetic polymorphisms (SNPs) of glutathione transferases (GSTM1, GSTP1, GSTT1), catalase gene (CAT, rs7943316), sirtuin 1 gene (SIRT1, rs10823108), acetylcholinesterase gene (ACHE, rs2571598), and butyrylcholinesterase gene (BCHE, rs3495) were screened in the OP-exposed individuals to find the possible causative association with oxidative stress and toxicity. Cholinesterase and antioxidant activities were measured by colorimetric methods using a spectrophotometer. Salting-out method was employed for DNA extraction from blood followed by restriction fragment length polymorphism (RFLP) for molecular analysis. Cholinergic enzymes were significantly decreased in OP-exposed groups. Catalase and SOD were decreased and MDA and FRAP were increased in OP-exposed groups compared to unexposed groups in both groups. GSH was decreased only in Pakistani OPs-exposed group. Molecular analysis of ACHE, BCHE, Catalase, GSTP1, and GSTM1 SNPs revealed a tentative association with their phenotypic expression that is level of antioxidant and cholinergic enzymes. The study concludes that chronic OPs exposure induces oxidative stress which is associated with the related SNP polymorphism. The toxicogenetics of understudied SNPs were examined for the first time to our understanding. The findings may lead to a newer area of investigation on OPs induced health issues and toxicogenetics.

Nano-interface driven electrochemical sensor for pesticides detection based on the acetylcholinesterase enzyme inhibition

Forensic Science Laboratories usually receive numerous cases of suicidal, accidental, and homicidal poisoning most often involving organophosphorus (OP) pesticides. The toxicity of the OP pesticides arises due to their ability to inhibit the activity of acetylcholinesterase (AChE), a cholinergic enzyme that is essential for the proper functioning of the central nervous system. Conventional techniques which are currently in use for pesticide detection are time-consuming, need upskilled technicians as well as suffer from low sensitivity. Therefore, the more rapid and sensitive electrochemical biosensors based on the principle of AChE enzyme inhibition have emerged out to be a simple and promising alternative to the conventional techniques. Since, most of the time, the poison isolated from biological material in poisoning cases is in nM quantities, an attempt has been made for the development of biosensor with enhanced sensitivity in the nM range using reduced graphene oxide (rGO) and zinc oxide nanoflowers (ZnONFs). The rGO and ZnONFs were synthesized chemically and deposited electrochemically on the Au electrode. AChE was immobilized onto this prepared nano-interface (ZnONFs/rGO/Au) through chitosan and glutaraldehyde cross-linking. The fabricated sensor was characterized step by step with cyclic voltammogram and electrochemical impedance spectroscopy. This advanced nanomaterials based techniques has been explored for detecting pesticides in visceral samples. The limit of detection (LOD) for the present sensor was 0.01 nM for OP pesticides.

Shaddock (Citrus maxima) peels extract restores cognitive function, cholinergic and purinergic enzyme systems in scopolamine-induced amnesic rats

This study investigated the effect of shaddock peels extract on cognitive function in scopolamine-induced amnesic rats. Wistar rats were pretreated with shaddock peels extract (50 and 100 mg/kg) and donepezil (5 mg/kg) for fourteen days via oral administration. Memory impairment was induced at the end of the treatment period via a single intraperitoneal administration of scopolamine (3 mg/kg). Thereafter, the animals were subjected to behavioral studies (Morris water maze and Y-maze tests). Finally, the rats were sacrificed and the hippocampus of the rat's brain was isolated for biochemical analyses. The results showed a significant decrease in memory and cognitive function as revealed by Morris water maze and Y-maze tests in scopolamine-induced rats which were reversed by shaddock peels extract. Also, there was a significant decrease in the activity of adenosine monophosphohydrolase (AMPase) with a simultaneous increase in activities of adenosine deaminase (ADA), adenosine triphosphate diphosphohydrolase (ATPdase), acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in scopolamine-induced rats when compared with the control. Besides, a significant increase in malondialdehyde (MDA) and reactive oxygen species (ROS) levels were observed in scopolamine-induced rats. However, donepezil or shaddock peels extract (50 and 100 mg/kg) caused a significant inhibitory effect on AChE, and ADA activities when compared to scopolamine-induced rats. Rats treated with shaddock peels extract also showed a significant reduction in MDA and ROS levels compared to scopolamine-induced rats. Therefore, our findings showed that the cognitive-enhancing effects of shaddock peels extract could be due to antioxidant activities and modulation of some enzymes linked with cognitive dysfunction.

Naringin attenuates Bisphenol-A mediated neurotoxicity in hypertensive rats by abrogation of cerebral nucleotide depletion, oxidative damage and neuroinflammation

We examined whether active fruit naringin can reduce the risk of BPA-mediated neurotoxicity in L-NAME induced hypertensive rats and whether the modulation could be linked to improvement of brain NO signaling. Male albino rats were randomly distributed into eight (n = 7) groups. Group I was control animals, Group II was orally-treated with L-NAME, Group III was orally treated with 100 mg/kg BPA, Group IV was orally-treated with L-NAME +100 mg/kg BPA. Group V was orally-administered with L-NAME +80 mg/kg NAR. Group VI was orally-administered with 100 mg/kg BPA +80 mg/kg NAR. Group VII was orally-administered with L-NAME+100 mg/kg BPA +80 mg/kg NAR. Lastly, group VIII was orally-treated with 80 mg/kg NAR. The treatment lasted for 14 days. Sub-acute exposure to L-NAME and BPA induced hypertension and mediated-neuroinflammation at CA-2 and CA-4 of hippocampus cells. It was evident by increase in PDE-51 and enzymes of ATP hydrolysis (ATPase, ADPase and AMPase) with corresponding upsurge in cholinergic (AChE and BuChE), dopaminergic (MAO-A) and adenosinergic (ADA) enzymes as well as movement disorder. The hypertensive-mediated neurotoxicity was related to alteration of NO signaling and higher release of pro-inflammatory cytokines (TNF-α and IL-1β), apoptotic proteins (P53 and caspace-9) and facilitated entry of T-lymphocytes (CD43+) into CNS through blood brain barrier potentiated by antigen presenting cells. Hence, these features of BPA-mediated neurotoxicity in L-NAME induced hypertensive rats were prohibited by co-administration of NAR through production of neuro-inflammatory mediators, stabilizing neurotransmitter enzymes, normalizing NO signaling and improving brain histology.

Enteromorpha prolifera Extract Improves Memory in Scopolamine-Treated Mice via Downregulating Amyloid-β Expression and Upregulating BDNF/TrkB Pathway.

Enteromorpha prolifera, a green alga, has long been used in food diets as well as traditional remedies in East Asia. Our preliminary study demonstrated that an ethyl acetate extract of Enteromorpha prolifera (EAEP) exhibited the strongest antioxidant activity compared to ethanol or water extracts. Nonetheless, there has been no report on the effect of EAEP on memory impairment due to oxidative damage. This study investigated whether EAEP could attenuate memory deficits in an oxidative stress-induced mouse model. EAEP was orally administered (50 or 100 mg/kg body weight (b.w.)) to mice and then scopolamine was administered. The oral administration of EAEP at 100 mg/kg b.w. significantly restored memory impairments induced by scopolamine, as evaluated by the Morris water maze test, and the passive avoidance test. Further, EAEP upregulated the protein expression of BDNF, p-CREB, p-TrkB, and p-Akt. Moreover, EAEP downregulated the expression of amyloid-β, tau, and APP. The regulation of cholinergic marker enzyme activities and the protection of neuronal cells from oxidative stress-induced cell death in the brain of mice via the downregulation of amyloid-β and the upregulation of the BDNF/TrkB pathway by EAEP suggest its potential as a pharmaceutical candidate to prevent neurodegenerative diseases.

Possible Correlation between Cholinergic System Alterations and Neuro/Inflammation in Multiple Sclerosis.

Multiple sclerosis (MS) is an autoimmune and demyelinating disease of the central nervous system. Although the etiology of MS is still unknown, both genetic and environmental factors contribute to the pathogenesis of the disease. Acetylcholine participates in the modulation of central and peripheral inflammation. The cells of the immune system, as well as microglia, astrocytes and oligodendrocytes express cholinergic markers and receptors of muscarinic and nicotinic type. The role played by acetylcholine in MS has been recently investigated. In the present review, we summarize the evidence indicating the cholinergic dysfunction in serum and cerebrospinal fluid of relapsing–remitting (RR)-MS patients and in the brains of the MS animal model experimental autoimmune encephalomyelitis (EAE). The correlation between the increased activity of the cholinergic hydrolyzing enzymes acetylcholinesterase and butyrylcholinesterase, the reduced levels of acetylcholine and the increase of pro-inflammatory cytokines production were recently described in immune cells of MS patients. Moreover, the genetic polymorphisms for both hydrolyzing enzymes and the possible correlation with the altered levels of their enzymatic activity have been also reported. Finally, the changes in cholinergic markers expression in the central nervous system of EAE mice in peak and chronic phases suggest the involvement of the acetylcholine also in neuro-inflammatory processes.

Expression profiling of precuneus layer III cathepsin D-immunopositive pyramidal neurons in mild cognitive impairment and Alzheimer's disease: Evidence for neuronal signaling vulnerability.

The precuneus (PreC; Brodmann area 7), a key hub within the default mode network (DMN) displays amyloid and tau-containing neurofibrillary tangle (NFT) pathology during the onset of Alzheimer's disease (AD). PreC layer III projection neurons contain lysosomal hydrolase cathepsin D (CatD), a marker of neurons vulnerable to NFT pathology. Here we applied single population laser capture microdissection coupled with custom-designed microarray profiling to determine the genetic signature of PreC CatD-positive-layer III neurons accrued from postmortem tissue obtained from the Rush Religious Orders Study (RROS) cases with a premortem clinical diagnosis of no cognitive impairment (NCI), mild cognitive impairment (MCI) and AD. Expression profiling revealed significant differential expression of key transcripts in MCI and AD compared to NCI that underlie signaling defects, including dysregulation of genes within the endosomal-lysosomal and autophagy pathways, cytoskeletal elements, AD-related genes, ionotropic and metabotropic glutamate receptors, cholinergic enzymes and receptors, markers of monoamine neurotransmission as well as steroid-related transcripts. Pervasive defects in both MCI and AD were found in select transcripts within these key gene ontology categories, underscoring the vulnerability of these corticocortical projection neurons during the onset and progression of dementia. Select PreC dysregulated genes detected via custom-designed microarray analysis were validated using qPCR. In summary, expression profiling of PreC CatD -positive layer III neurons revealed significant dysregulation of a mosaic of genes in MCI and AD that were not previously appreciated in terms of their indication of systems-wide signaling defects in a key hub of the DMN.

Enteromorpha prolifera Extract Protects against Scopolamine‐induced Memory Impairment in Mice

Enteromorpha prolifera (EP) contains various nutrients and phytochemicals that have been reported to play role in antioxidant activity, hypolipidemic and immunomodulatory. However, the neuroprotective effect of EP has not been reported. The present study examined whether EP ethylacetate extract(EPE) improves memory impairment induced by scopolamine. EPE (50 or 100 mg/kg) was administered orally to mice for 14 days, and then scopolamine (2 mg/kg, i.p.) was administered together with EPE for another 14 days. Memory performance was evaluated using the Morris water maze test and passive avoidance test. Also, brain acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) activities, biomarkers of oxidative stress and the loss of neuronal cells in hippocampus was evaluated by histological examinations. Administration of EPE significantly restored memory impairments induced by scopolamine, as demonstrated by the Morris water maze test (escape latency and number of crossing platform area), and in the passive avoidance test. Treatment with EPE inhibited the AChE activity and increased the ChAT activity in the brain of memory impaired mice induced by scopolamine. Additionally, the administration of EPE significantly prevented the increase in lipid peroxidation and the decrease in GSH levels in brain of mice treated with scopolamine. Also, the EPE treatment restored the activities of antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR) to control level. Furthermore, scopolamine‐induced oxidative damage of neurons in hippocampal CA1 and CA3 regions were prevented by EPE treatment. It is suggested that EPE may be a useful for mitigating memory improvement via the regulation of cholinergic marker enzyme activities and the suppression of oxidative damage of neurons in the brain of mice treated with scopolamine.

Vanillin and vanillic acid modulate antioxidant defense system via amelioration of metabolic complications linked to Fe2+-induced brain tissues damage.

The therapeutic effect of phenolics on neurodegenerative diseases has been attributed to their potent antioxidant properties. In the present study, the neuroprotective activities of vanillin and vanillic acid were investigated in Fe2+- induced oxidative toxicity in brain tissues by investigating their therapeutic effects on oxidative imbalance, cholinergic and nucleotide-hydrolyzing enzymes activities, dysregulated metabolic pathways. Their cytotoxicity was investigated in hippocampal neuronal cell lines (HT22). The reduced glutathione level, SOD and catalase activities wereameliorated in tissues treated with the phenolics, with concomitant depletion of malondialdehyde and nitric oxide levels. They inhibited acetylcholinesterase and butyrylcholinesterase activities, while concomitantly elevated ATPase activity. Treatment with vanillin led to restoration of oxidative-depleted metabolites and reactivation of the pentose phosphate and purine metabolism pathways, with concomitant activation of pathways for histidine and selenoamino metabolisms. While vanillic acid restored and reactivated oxidative-depleted metabolites and pathways but did not activate any additional pathway. Both phenolics portrayed good bindingaffinity for catalase, with vanillic acid havingthe higher binding energy of -7.0kcal/mol. Both phenolics were not cytotoxic on HT22 cells, and their toxicity class were predicted to be 4. Only vanillin was predicted to be permeable across the blood brain barrier (BBB). These results insinuate that vanillin and vanillic acid confer a neuroprotective effect on oxidative brain damage, when vanillin being the most potent.

Fucoidan serves a neuroprotective effect in an Alzheimer's disease model.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder that causes memory and cognitive deficits. The present study was carried out to evaluate the protective effects of fucoidan in monocrotophos induced AD in Drosophila melanogaster. In silico studies showed that fucoidan exhibited binding energy of -9.3 kcal with proteins. Consistent with this, fucoidan, in a dose and time-dependent fashion, had inhibitory activity against cholinergic and monoamine-metabolized enzymes in vitro. Fucoidan inhibited the increase in total mRNA and protein in monocrotophos fed flies and prevented changes in biochemicals, neurochemicals and latency time of locomotor, learning and memory induced by monocrotophos. Together, the findings show that fucoidan serves a neuroprotective effect in Alzheimer's disease model in D. melanogaster.

Tetrahydroquinoline-Isoxazole/Isoxazoline Hybrid Compounds as Potential Cholinesterases Inhibitors: Synthesis, Enzyme Inhibition Assays, and Molecular Modeling Studies.

A series of 44 hybrid compounds that included in their structure tetrahydroquinoline (THQ) and isoxazole/isoxazoline moieties were synthesized through the 1,3-dipolar cycloaddition reaction (1,3-DC) from the corresponding N-allyl/propargyl THQs, previously obtained via cationic Povarov reaction. In vitro cholinergic enzymes inhibition potential of all compounds was tested. Enzyme inhibition assays showed that some hybrids exhibited significant potency to inhibit acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Especially, the hybrid compound 5n presented the more effective inhibition against AChE (4.24 µM) with an acceptable selectivity index versus BChE (SI: 5.19), while compound 6aa exhibited the greatest inhibition activity on BChE (3.97 µM) and a significant selectivity index against AChE (SI: 0.04). Kinetic studies were carried out for compounds with greater inhibitory activity of cholinesterases. Structure–activity relationships of the molecular hybrids were analyzed, through computational models using a molecular cross-docking algorithm and Molecular Mechanics/Generalized Born Surface Area (MM/GBSA) binding free energy approach, which indicated a good correlation between the experimental inhibition values and the predicted free binding energy.

Impacts of Escherichia coli infection in young breeder chicks on the animal behavior and cerebral activity of purinergic and cholinergic enzymes involved in the regulation of molecules with neurotransmitter and neuromodulator function

The objective of this study was to evaluate if infection by Escherichia coli in juvenile breeder chicks alters the activity of enzymes involved in neurotransmission and cerebral immunomodulation, including acetylcholinesterase (AChE), nucleoside triphosphate diphosphohydrolase (NTPDase), 5′-nucleotidase (5′NT) and adenosine deaminase (ADA), as well as their effects on the pathogenesis of the disease. We divided 20 growing breeder chicks into two groups (n = 10 per group). One group was experimentally infected with 1 mL of culture medium containing 1 × 108 CFU of E. coli intraperitoneally. The other was the negative control. On the tenth day after infection, the animals were euthanized and brain samples were collected. Macroscopically, pericarditis and hepatic congestion were observed in the birds, but without histopathological lesions in the encephalon although the bacterium was present in the cerebral cortex of all animals in the infected group (i.e., they were PCR-positive). The activity of AChE, NTPDase, 5′-NT and ADA were evaluated in the cerebral homogenates of the birds after 10 days of infection. AChE activity in the cerebral cortex was lower in the infected group than in the control; there was an increase in the activity of NTPDase, 5′-nucleotidase and ADA, possibly indicating greater hydrolysis of ATP (P < 0.001), ADP (P < 0.01) and AMP (P < 0.01), followed by increased adenosine deamination (P < 0.001). Despite these changes, no apparently diseased animals were observed throughout the experimental period. Therefore, such changes in enzymatic activity may affect the functioning of the central nervous system because these enzymes are responsible for extracellular regulation of molecules that act on neurotransmission and immunomodulation such as acetylcholine, ATP and adenosine.

Thyme (Thymus vulgaris) leaf extract modulates purinergic and cholinergic enzyme activities in the brain homogenate of 5-fluorouracil administered rats

This study investigated the effect of Thyme (Thymus vulgaris) leaf aqueous extract (TE) on purine and acetylcholine hydrolyzing enzyme activities and antioxidant status in the brain of 5-fluorouracil (5-FLU) administered rats. Sixty adult Wistar rats (210–225 g) were divided into Ten groups (n = 6). Group 1: received normal saline orally (NC), group 2, 3, 4 and 5 rats were respectively received 100, 200 and 400 mg/kg of TE and Vitamin C (25 mg/kg) orally; group 6 rats received normal saline orally and 150 mg/kg of 5-FLU via i.p., on the last day; rats in group 7, 8, 9 and 10 were orally pretreated with 100, 200 and 400 mg/kg of TE and Vit C (25 mg/kg) respectively, and administered with 150 mg/kg of 5-FLU i.p., on the last day. The purine [adenosine 5'-tri- and di-phosphatases (ATPase and ADPase), adenosine deaminase (ADA)] nucleotide and acetylcholine hydrolyzing [acetylcholinesterase (AChE) and butrylcholinesterase (BChE)] enzymes as well as arginase activities and antioxidant status were assessed. The result revealed that untreated 5-FLU rats had increased enzyme activities, which was reduced in the normal rats treated TE compared to NC. Interestingly, the TE-pretreated rats administered with 5-FLU had reduced enzyme activities compared with untreated 5-FLU rats. The antioxidant status was also normalized in the 5-FLU administered rats pretreated with TE Thus, the neuroprotective mechanisms of TE could be linked to attenuation of neuroactive enzymes and antioxidant status.