Plasma Butyrylcholinesterase Research Articles

Protective effect of dietary supplements against streptozotocin-induced Alzheimer’s disease in mice

Introduction: Alzheimer’s disease (AD) is a neurodegenerative problem that is increased progressively due to the increment of aging worldwide. Phytochemicals play an important role in the protection from neurodegeneration. The present study aimed to evaluate the protective effect of two dietary supplements (DS) rich in betalains, anthocyanins, and omega-3 fatty acids against AD. Methods: Two dietary supplements (DS I and DS II) were prepared; the first one was a mixture of anthocyanin-rich extract of purple carrot and flaxseed oil (DS I), while the second was a mixture of betalains-rich extract of beetroot and flaxseed oil (DS II). The protective effects of both DS were evaluated in an AD model. AD was induced in mice by intracerebroventricular (ICV) injection of streptozotocin (STZ) (3 mg/kg). Biochemical changes in brain tissue and plasma were determined. Behavioral of mice was evaluated through Y–maze test, Morris water maze, and novel object recognition test. Changes in brain tissues were assessed through histopathological examination. In vitro antioxidant activities of DS I and DS II were evaluated. Also, the contents of total phenolics, anthocyanins, betalains, and fatty acids profile were assessed. Results: Both DS investigated in the present study showed significant improvement (P < 0.05) in acetylcholinesterase, antioxidant enzymes, tumor necrosis factor-α (TNF-α) and malondialdehyde (MDA)in brain tissue and butyrylcholinesterase in plasma in association with amelioration in the behavioral tests and histopathological changes of the brain tissue. Conclusion: Both DS showed protective effects against STZ induced AD in mice due to the presence of anthocyanins, betalains, and omega-3 fatty acids.

Multifarious Functions of Butyrylcholinesterase in Neuroblastoma: Impact of BCHE Deletion on the Neuroblastoma Growth In Vitro and In Vivo.

The physiological functions of butyrylcholinesterase (BChE) and its role in malignancy remain unexplained. Our studies in children newly diagnosed with neuroblastoma indicated that BChE expressions is proportional to MYCN amplification suggesting that pathogenesis of high-risk disease may be related to the persistent expression of abnormally high levels of tumor-associated BChE. BChE-deficient neuroblastoma cells (KO [knockout]) were produced from MYCN -amplified BE(2)-C cells (WT [wild-type]) by the CRISPR-Cas9 targeted disruption of the BCHE locus. KO cells have no detectable BChE activity. The compensatory acetylcholinesterase activity was not detected. The average population doubling time of KO cells is 47.0±2.4 hours, >2× longer than WT cells. Reduced proliferation rates of KO cells were accompanied by the loss of N-Myc protein and a significant deactivation of tyrosine kinase receptors associated with the aggressive neuroblastoma phenotype including Ros1, TrkB, and Ltk. Tumorigenicity of WT and KO cells in male mice was essentially identical. In contrast, KO xenografts in female mice were very small (0.37±0.10 g), ~3× smaller compared with WT xenografts (1.11±0.30 g). Unexpectedly, KO xenografts produced changes in plasma BChE similarly to WT tumors but lesser in magnitude. The disruption of BCHE locus in MYCN -amplified neuroblastoma cells decelerates proliferation and produces neuroblastoma cells that are less aggressive in female mice.

Identification and Study of Biomarkers from Novichok-Inhibited Butyrylcholinesterase in Human Plasma.

To identify biomarkers of ethyl (1-(diethylamino)ethylidene)phosphoramidofluoridate (A234)- or methyl (1-(diethylamino)ethylidene)phosphoramidofluoridate (A232)-inhibited butyrylcholinesterase (BChE), we investigated nonapeptide adducts containing the active site serine, which plays a key role in enzyme activity, using LC-MS/HRMS. Biomarkers were acquired as expected, and they exhibited a significant amount of fragment ions from the inhibiting agent itself, in contrast to the MS2 spectra of conventional nerve agents. These biomarkers had a higher abundance of [M+2H]2+ ions than [M+H]+ ions, making doubly charged ions more suitable for trace analysis.

Kinetics of human butyrylcholinesterase inhibition by 1,9-dimethyl-methylene blue

Alzheimer’s disease (AD) is an irreversible and progressive neurodegenerative disorder, characterized by β-amyloid plaques, neurofibrillary tangles and loss of cholinergic neurons. Butyrylcholinesterase (BChE) inhibition is one of the most critical strategy for the treatment of AD since BChE causes inactivation of neurotransmitter acetylcholine and has positive effects on promoting the formation of β-amyloid fibrils. Our previous studies showed that various phenothiazine-derived compounds such as thionine and toluidine blue O (TBO) cause a potent inhibition of human cholinesterases. TBO was also found to reduce amyloid precursor protein processing in-vitro and in-vivo models of AD. In this study, it was aimed to determine the inhibitory effect of 1,9-dimethyl-methylene blue (DMMB), a phenothiazine-derived compound, on human plasma BChE and explore its inhibitory mechanism. The inhibition of human BChE was assessed by the colorimetric method of Ellman using butyrylthiocholine as substrate and 0-0.375 μM of DMMB. The kinetic findings showed that DMMB acts as a linear mixed-type inhibitor of human BChE with Ki value of 23 ± 0.004 nM and α= 3.6 ± 1.6. It was concluded that DMMB, which is a potent inhibitor effective at nM level may be helpful in designing new cholinesterase inhibitors for the treatment of AD.

Design and synthesis of novel tacrine-indole hybrids as potential multitarget-directed ligands for the treatment of Alzheimer's disease.

The authors report on the synthesis and biological evaluation of new compounds whose structure combines tacrine and indole moieties. Tacrine-indole heterodimers were designed to inhibit cholinesterases and β-amyloid formation, and to cross the blood-brain barrier. The most potent new acetylcholinesterase inhibitors were compounds 3c and 4d (IC50=25 and 39nM, respectively). Compound 3c displayed considerably higher selectivity for acetylcholinesterase relative to human plasma butyrylcholinesterase in comparison to compound 4d (selectivity index: IC50 [butyrylcholinesterase]/IC50 [acetylcholinesterase] =3 and 0.6, respectively). Furthermore, compound 3c inhibited β-amyloid-dependent amyloid nucleation in the yeast-based prion nucleation assay and displayed no dsDNA destabilizing interactions with DNA. Compounds 3c and 4d displayed a high probability of crossing the blood-brain barrier. The results support the potential of 3c for future development as a dual-acting therapeutic agent in the prevention and/or treatment of Alzheimer's disease.

21-Day dermal exposure to aircraft engine oils: effects on esterase activities in brain and liver tissues, blood, plasma, and clinical chemistry parameters for Sprague Dawley rats

ABSTRACT This dermal study tested the potential toxicity of grade 3 (G3) and 4 (G4) organophosphate-containing aircraft engine oils in both new (G3-N, G4-N) and used states (G3-U, G4-U) to alter esterase activities in blood, brain and liver tissues, clinical chemistry parameters, and electrophysiology of hippocampal neurons. A 300 µl volume of undiluted oil was applied in Hill Top Chamber Systems®, then attached to fur-free test sites on backs of male and female Sprague Dawley rats for 6 hr/day, 5 days/week for 21 days. Recovery rats received similar treatments and kept for 14 days post-exposure to screen for reversibility, persistence, or delayed occurrence of toxicity. In brain, both versions of G3 and G4 significantly decreased (32–41%) female acetylcholinesterase (AChE) activity while in males only G3-N and G4-N reduced (33%) AChE activity. Oils did not markedly affect AChE in liver, regardless of gender. In whole blood, G3-U decreased female AChE (29%) which persisted during recovery (32%). G4-N significantly lowered (29%) butyrylcholinesterase (BChE) in male plasma, but this effect was resolved during recovery. For clinical chemistry indices, only globulin levels in female plasma significantly increased following G3-N or G4-N exposure. Preliminary electrophysiology data suggested that effects of both versions of G3 and G4 on hippocampal function may be gender dependent. Aircraft maintenance workers may be at risk if precautions are not taken to minimize long-term aircraft oil exposure.

Inhibition of cholinesterases by safranin O: Integration of inhibition kinetics with molecular docking simulations

In the present study, the inhibitory mechanisms and effects of a synthetic phenazine dye, safranin O (SO) on human plasma butyrylcholinesterase (BChE), human erythrocyte acetylcholinesterase (AChE) and recombinant BChE mutants were investigated. Kinetic studies showed the following information: SO leaded to linear competitive inhibition of human plasma BChE with Ki = 0.44 ± 0.085 μM; α = ∞. It acted as a hyperbolic noncompetitive inhibitor of human erythrocyte AChE with Ki = 0.69 ± 0.13; α = 1; β = 0.08 ± 0.02. On the other hand, the inhibitory effects of SO on two BChE mutants, where A328 was modified to either F or Y, revealed differences in terms of inhibitory patterns and Ki values, compared to the obtained results with recombinant wild type BChE. SO was found to act as a linear competitive inhibitor of A328F and A328Y BChE mutants. Compared to recombinant wild type BChE, A328Y and A328F BChE mutants caused a 4- and 10-fold decrease in Ki value for SO, respectively. These findings were supported by molecular modelling studies. In conclusion, SO is a potent inhibitor of human cholinesterases and may be useful in the design and development of new drugs for the treatment of AD.

A single dose of the organophosphate triazophos induces fear extinction deficits accompanied by hippocampal acetylcholinesterase inhibition

Acute organophosphate (OP) poisoning, particularly by suicide attempts, generates high mortality and morbidity. Few studies have systematically addressed the consequences of acute OP intoxication on cognition and memory of survivors. Preclinical evidence suggests that acute OP-induced effects are associated with inhibiting the brain acetylcholinesterase (AChE) enzyme. The OP triazophos has been used worldwide, although its effects on mnemonic processing are yet to be investigated. Based on the above, the present study investigated whether acute triazophos intoxication interferes with the expression and extinction of contextual fear memory in rats. Hippocampal and amygdalar AChE activity and plasma butyrylcholinesterase (BChE) were measured at the end of the experiment to confirm the cholinergic overstimulation. Independent cohorts of animals intoxicated with triazophos were evaluated in the novel object recognition (NOR) test, a less aversive associative memory task. At the dose of 15 mg/kg, triazophos administered immediately after contextual fear conditioning impaired the extinction but not the expression of freezing behavior. Triazophos poisoning induced no changes in the discrimination index in the NOR test. Triazophos inhibited the AChE activity in a time- and brain region-dependent manner. Our findings suggest that fear memory extinction deficits induced by acute triazophos intoxication are accompanied by hippocampal AChE inhibition. The deficient fear extinction associated with acute OP poisoning may represent a behavioral and biochemical phenotype helpful to study mechanisms of neurotoxicity and treatment approach of OP suicide survivors.

Monoclonal antibodies to fetal bovine serum acetylcholinesterase distinguish between acetylcholinesterases from ruminant and non-ruminant species

Two types of cholinesterases (ChEs) are present in mammalian blood and tissues: acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). While AChE regulates neurotransmission by hydrolyzing acetylcholine at the postsynaptic membranes and neuromuscular junctions, BChE in plasma has been suggested to be involved in detoxifying toxic compounds. This study was undertaken to establish the identity of circulating ChE activity in plasmas from domestic animals (bovine, ovine, caprine, porcine and equine) by assessing sensitivity to AChE-specific inhibitors (BW284c51 and edrophonium) and BChE-specific inhibitors (dibucaine, ethopropazine and Iso-OMPA) as well as binding to anti-FBS AChE monoclonal antibodies (MAbs). Based on the inhibition of ChE activity by ChE-specific inhibitors, it was determined that bovine, ovine and caprine plasma predominantly contain AChE, while porcine and equine plasma contain BChE. Three of the anti-FBS AChE MAbs, 4E5, 5E8 and 6H9, inhibited 85–98% of enzyme activity in bovine, ovine and caprine plasma, confirming that the esterase in these plasmas was AChE. These MAbs did not bind to purified recombinant human or mouse AChE, demonstrating that these MAbs were specific for AChEs from ruminant species. These MAbs did not inhibit the activity of purified human BChE, or ChE activity in porcine and equine plasma, confirming that the ChE in these plasmas was BChE. Taken together, these results demonstrate that anti-FBS AChE MAbs can serve as useful tools for distinguishing between AChEs from ruminant and non-ruminant species and BChEs.

Decreased Activity of Circulating Butyrylcholinesterase in Blood Is an Independent Prognostic Marker in Pancreatic Cancer Patients.

Introduction: The activity of butyrylcholinesterase (BChE) in blood reflects liver function and has recently been associated with systemic inflammatory response and tumor cachexia. As these conditions have been previously linked with pancreatic cancer (PC), the purpose of the present study was to evaluate the prognostic impact of plasma BChE in PC. Methods: Data from 574 consecutive PC patients, treated between 2004 and 2018 at a single academic center, was evaluated. The primary endpoint was cancer-specific survival (CSS), analyzed by Kaplan–Meier curve, and both univariate and multivariate Cox proportional models. Results: BChE activity negatively correlated with other liver parameters (bilirubin, gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and C-reactive protein (CRP)), and positively correlated with albumin levels, respectively (p < 0.01). In univariate analysis, a low plasma BChE activity was a factor of poor CSS (hazard ratio: 1.4, 95% confidence interval: 1.129–1.754, p = 0.002). In multivariate analysis, tumor stage, tumor grade, administration of chemotherapy, bilirubin levels and a low BChE activity (hazard ratio: 1.42, 95% confidence interval: 1.10–1.82; p = 0.006) were identified as independent prognostic factors. Conclusion: Decreased activity of BChE in blood plasma predicts shorter survival time in PC patients. Therefore, BChE might be helpful in additional stratification of patients into different prognostic risk groups.

Correlation of plasma butyrylcholinesterase concentration with Acethylcholinesterase H353N polymorphism in the inflammatory response of Chagas disease patients

Trypanosoma cruzi infection triggers an intense production of pro-inflammatory cytokines mediated by T helper 1 lymphocytes, inducing the anti-inflammatory reflex of acetylcholine (ACh). The ACh concentration modulation is associated to the two major esterases, the acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). AChE H353N protein polymorphism is related to low Chagas chronic disease prognostic. In order to evaluate the correlation of plasmatic BuChE concentration and the presence of AChE H353N polymorphism in Chagas disease patients and healthy individuals, we studied two groups of individuals, one of 61 Chagas disease patients and another of 74 healthy individuals. Plasma concentration of BuChE was measured by the chemiluminescent method and AChE H353N polymorphism was investigated by PCR-RFLP and sequencing of the respective encoding AChE gene fragment. The BuChE concentration was statistically higher in Chagas disease patients, with no AChE genotype significant influence. AChE genotypes YT*A/YT*A, YT*A/YT*B and YT*B/YT*B, respectively, were expressed in 53 (86.88%), 7 (11.46%) and one (1.64%) chagasic patients, and in 68 (91.89%), 6 (8.10%) and none healthy individuals. BuChE activity may represent an important marker for chronic Chagas disease inflammatory process and prognostic. Lower BuChE concentration correlated with AChE YT*B allele, although without statistical power.

Review of Biomarkers and Analytical Methods for Organophosphate Pesticides and Applicability to Nerve Agents.

Recent malicious use of chemical warfare agents (CWAs) is a reminder of their severity and ongoing threat. One of the main categories of CWAs is the organophosphate (OP) nerve agents. Presently, there is an urgent need to identify and evaluate OP nerve agent biomarkers that can facilitate identification of exposed individuals post-CWA incident. While exposures to OP nerve agents may be scenario-specific, the public is commonly exposed to OP compounds through the ubiquitous use of OP pesticides, which are chemically related to nerve agents. Therefore, a systematic literature review and methodological quality assessment were conducted for OP pesticide biomarker studies to serve as a baseline to assess if these approaches may be adapted to OP nerve agent exposures. We conducted a systematic literature review to identify biomarkers of OP pesticide exposures. English language studies of any design that reported primary data on biomarkers for exposures in nonhuman primates or adult human study participants were eligible for inclusion. Using standard criteria for assessing the completeness of reported analytical methods, the quality of study methods was critically evaluated. A total of 1,044 studies of biomarkers of OP pesticide exposure were identified, of which 75 articles satisfied the inclusion and exclusion criteria. These studies described 143 different analyte/sample matrix combinations: 99 host-based biomarkers, 28 metabolites, 12 pesticides, and 4 adducts. The most commonly reported biomarkers were dialkyl phosphate urinary metabolites (22 studies), blood acetylcholinesterase, and plasma butyrylcholinesterase (26 studies each). None of the assessed quality review criteria were fully addressed by all identified studies, with almost all criteria scoring less than 50%. Cholinesterase activity may have utility for identifying individuals with exposures surpassing a given threshold of OP nerve agent, but further investigation of how acetylcholinesterase and butyrylcholinesterase levels correlate with observed patient symptoms may be required to ensure accuracy of results. As CWAs and nerve agents are more readily used, more standardized reporting of biomarker measurements are needed to develop new approaches for OP nerve agent biomarkers.

Fish Oil and Primrose Oil Suppress the Progression of Alzheimer's Like Disease Induced by Aluminum in Rats.

The role of fish oil, primrose oil and their mixture in ameliorating the changes in Alzheimer's like model was evaluated in rats. Primrose oil and primrose/fish oil mixture fatty acids composition was assessed by gas chromatography. The rat experiment consisted of 5 groups; the first fed on balanced diet as control normal (CN); the other four groups treated with intraperitoneal aluminum lactate and consumed dyslipidemic diet; one group served as control Alzheimer's like disease (CA) while the other three groups (test groups) received daily oral dose from primrose oil, fish oil and primrose/fish oil mixture separately for 5 weeks. Results showed primrose oil and primrose/ fish oil mixture to contain gamma linolenic acid as 9.15 and 4.3% of total fatty acids, respectively. Eicosapentaenoic and docosahexaenoic were present as 10.9 and 6.5 %, respectively in the oil mixture. Dyslipidemia and increased erythrocyte sedimentation rate (ESR), plasma butyrylcholinesterase (BChE), brain malondialdehyde (MDA) and NO with decrease in plasma magnesium, brain catalase, reduced glutathione, body weight gain and brain weight were demonstrated in CA compared to CN. Brain histopathology and immuno-histochemistry showed neuronal degeneration and neurofibrillary tangles with elevated myeloperoxidase and nuclear factor-kappa B in CA compared to CN. The tested oils demonstrated neuro-protection reflected in the variable significant improvement of biochemical parameters, immuno-histochemistry and brain histopathology. Primrose/fish oil mixture was superior in reducing ESR, brain MDA, plasma activity of BChE and brain histopathological changes along with elevating plasma magnesium. Primrose/fish oil mixture and fish oil were more promising in improving plasma high density lipoprotein cholesterol (HDL-C) than primrose. Fish oil was the most efficient in improving plasma total cholesterol (T-C), low density lipoprotein cholesterol and T-C /HDL-C. Primrose/fish oil mixture and primrose oil were superior in elevating brain catalase compared to fish oil. Other parameters were equally improved by the different oil treatments. Primrose oil, fish oil and their mixture reduced the progression of Alzheimer's disease in rats with superiority to primrose/fish oil mixture.

Assessment of DNA damage and cholinesterase activity in soybean farmers in southern Brazil: High versus low pesticide exposure

The aim of this study was to evaluate the DNA damage in soybean growers during two agricultural periods of a crop season (high and low exposure) and a control group, as well as butyrylcholinesterase (BChE) activity during these exposure periods in order to estimate the degree of BChE inhibition for the exposed group. DNA damage in peripheral whole blood was evaluated by the comet assay and plasma BChE activity was accessed as a measure of exposure to cholinesterase inhibitors. None of the soybean growers reported using full Personal Protective Equipment (PPE). BChE was lower in high exposure period than in low exposure period and DNA damage index was significantly increased in the high exposure period than in the low exposure period. In addition, DNA damage in both exposure periods was higher than control group. No correlation was found between exposure time and DNA damage and BChE activity. However, negative correlation was observed between DNA damage in high and low exposure periods. The results indicate that soybean growers are exposed to cholinesterase inhibitors and to pesticides mixtures with genotoxic potential.

A pilot clinical study of the neuromuscular blocker rocuronium to reduce the duration of ventilation after organophosphorus insecticide poisoning

Background: A common manifestation of organophosphorus insecticide self-poisoning is prolonged respiratory failure due to neuromuscular junction dysfunction and likely nicotinic receptor overstimulation. We aimed at collecting preliminary data on whether addition of the competitive nicotinic antagonist rocuronium to standard early therapy might be clinically feasible and associated with reduced duration of ventilation.Methods: A pilot three-arm dose–response phase II trial was set up to compare bolus doses of rocuronium bromide titrated to produce initial >95% or 50% inhibition of neuromuscular function, measured using acceleromyography, plus standard treatment, versus standard treatment alone. After attaining inhibition, patients receiving bolus rocuronium then received rocuronium infusions for a maximum of 120 h. Primary outcome was duration of intubation; secondary outcome was case fatality. Plasma butyrylcholinesterase activity was measured throughout the inpatient stay. Blood was analysed to confirm the organophosphorus insecticide ingested.Results: Forty-five patients were randomised to receive: rocuronium to initially attain 95% inhibition (Roc>95, n = 15), rocuronium to initially attain 50% inhibition (Roc50, n = 14), or no rocuronium (control, n = 16). The most commonly ingested pesticide was profenofos (29/45, 64.4%). Butyrylcholinesterase activity remained severely inhibited for the duration of the study for most patients. Case fatality was 9/45 (20%) and similar across study arms: control 3/16 (18.8%), Roc50 4/14 (28.6%) and Roc>95 2/15 (13.3%) (p = .5842). When excluding patients who died, median [IQR] duration of intubation was significantly longer in the Roc50 (259.5 [176–385] h) and Roc>95 (226.8 [186–355] h) groups compared to controls (88.5 [47–160] h, p = .0162 and p = .0016, respectively).Conclusions: In this pilot dose–response study, we found no evidence that rocuronium in addition to standard therapy reduced the duration of intubation. It is possible that it worsened neuromuscular junction function. Further clinical research, including testing of shorter duration regimens, needs to be performed before nicotinic antagonists can be used in the clinical management of OP poisoning.

Plasma cholinesterase is associated with Chinese adolescent overweight or obesity and metabolic syndrome prediction.

Purpose: To determine the plasma concentrations of butyrylcholinesterase (BChE), also known as pseudocholinesterase, in different weight categories of adolescents, and to explore the possible association between plasma BChE and overweight (OW), obesity, and metabolic syndrome (MetS) in Chinese adolescents.Patients and methods: This cross-sectional study included 1,236 Chinese adolescents (194 obese [OB], 188 OW, 732 normal weight [NW], and 122 underweight [UW]). The biochemical variables and anthropometric variables of the study participants were evaluated. Plasma BChE level was measured by DGKC method.Results: OB was associated with a higher prevalence of upper strata plasma BChE levels when compared with the BChE levels in UW, NW, and OW group. A logistic regression analysis showed that plasma BChE was positively associated with the OB group when compared with the NW group. Boys in the OW group, but not the OB group, had a significantly higher prevalence of upper stratum of BChE levels. Plasma triglyceride, total cholesterol, low-density lipoprotein-cholesterol, and ApoB levels were positively associated with the upper stratum of BChE levels when compared with lower stratum. MetS and most of its components were more prevalent among subjects with upper stratum rather than lower stratum BChE levels. Receiver operating characteristic curves for plasma BChE in subjects with MetS indicated that the AUC was 0.80 (95%CI:0.70–0.90,P<0.001) and 0.89 (95%CI:0.82–0.95,P<0.001) in girls and boys, respectively. After adjusting for age, the multivariate-adjusted odds ratio for MetS in the upper stratum of BChE levels was 8.73 (95%CI: 3.49–21.84) in the boys cohorts and also in the girls cohorts (OR=1.71, 95%CI: 1.35–21.70).Conclusion: This study confirmed an association between BChE levels and weight status in Chinese adolescents, and demonstrated that the upper strata of plasma BChE levels were associated with being OW, and even more highly associated with obesity. Plasma BChE levels were positively associated with MetS and its components and could be useful for identifying adolescents with MetS.

Oxidative and histopathological alterations after sub-acute exposure of diisopropyl phosphorofluoridate in mice: Beneficial effect of N‑acetylcysteine

AimsProtective efficacy of N‑acetylcysteine (NAC) was assessed against sub-acute diisopropyl phosphorofluoridate (DFP) poisoning in mice. Main methodsMice were allocated into nine groups of six each: vehicle control; DFP (0.125 LD50 ≈ 0.483 mg/kg bwt, s.c.); DFP + Atropine (ATR, 10 mg/kg bwt, i.p., 0 min); DFP + Pralidoxime (2-PAM, 30 mg/kg bwt, i.m., 0 min); DFP + NAC (150 mg/kg bwt, i.p., −60 min); DFP + ATR + NAC; DFP + 2-PAM + NAC; DFP + ATR + 2-PAM; and DFP + ATR + 2-PAM + NAC. Animals received various treatments for 21 d daily. Plasma butyrylcholinesterase (BChE) was measured after 7, 14 and 21 d of exposure. Brain acetylcholinesterase (AChE) and reduced glutathione (GSH), malondialdehyde (MDA), glutathione peroxidase (GPx), glutathione reductase (GR), catalase (CAT), and superoxide dismutase (SOD) were measured (brain, liver and kidney) after 21 d of exposure. Histopathology, immunohistochemistry, and Western blot for inducible nitric oxide synthase (iNOS) and c-fos were also performed. Key findingsDFP significantly reduced BChE and AChE levels. Diminished GSH, CAT, SOD (brain and liver), GPx, GR, and elevated MDA (Brain) levels were also observed. DFP caused notable histopathology (brain, liver and kidney) and over expression of iNOS, and c-fos proteins (brain). NAC enhanced the protective efficacy of ATR and 2-PAM in most parameters, without any appreciable protection in iNOS and c-fos expression. SignificanceNAC as an adjunct with ATR and 2-PAM, exhibited marked beneficial effects against sub-acute DFP poisoning, indicating its possible implications in the management of OP poisoning.

Assessment of genotoxicity and cholinesterase activity among women workers occupationally exposed to pesticides in tea garden

To assess the effects of exposure to pesticides in tea garden women workers, the present study evaluated the activities of erythrocyte acetylcholinesterase (AChE) and plasma butyrylcholinesterase (BChE) as biomarkers of pesticide exposure and DNA damage by comet assay as a biomarker of genotoxicity in peripheral blood lymphocytes. We examined 143 women subjects including 77 women workers exposed to pesticide in tea garden (exposed) and 66 women subject with no history of occupational exposure to pesticides (non-exposed). The two groups had similar mean ages, BMI and personal habits. The results revealed that exposed group had lower activity of AChE (2.86 ± 0.75 vs. 3.93 ± 0.87 U/ml) BChE (2.02 ± 0.74 vs. 2.60 ± 0.74 U/ml, p < 0.001) than those of non-exposed group. The mean comet parameters such as tail length (TL), % DNA in tail, tail moment (TM) and olive tail moment (OTM) used to measure DNA damage were significantly elevated in exposed workers compared to non-exposed (p < 0.001). There was no variation observed in Cholinesterase (ChEs) activity and Comet assay parameter based on age, personal habits, years of exposure and use of personal protective equipments (PPEs) except BChE activity. The multiple regression analysis showed no association of confounding factors on end points measured except slight negative association of BMI with tail length. Therefore this study suggests that the workers in tea garden might have exposed to pesticide through their occupation on a regular interval. Although this study could able to associate it with their occupational exposure, the role of other DNA damaging factors cannot be ruled out. Our study finding indicate the necessity of periodic monitoring of these biomarkers along with imparting education and training to minimise the risk.

Biochemical Analysis and Association of Butyrylcholinesterase SNPs rs3495 and rs1803274 with Substance Abuse Disorder.

Addiction is a complex mental and behavioral disorder that changes the neurochemistry and physiology of the brain. Genetics also plays a significant role in the pathophysiology of addiction. Butyrylcholinesterase (BChE), a cholinergic enzyme, has been implicated in the metabolism of various drugs, including cocaine, and an association between single-nucleotide polymorphisms (SNPs) of the butyrylcholinesterase gene (BCHE) and neuronal disorders has been reported. We report here the first investigation to be conducted on the status of BChE activity and the potential association of two BCHE gene SNPs, rs3495 (c.*189G > A) and rs1803274 (c.1699G>A, p.Ala567Thr, K-variant), with addiction vulnerability in heroin, hashish and polydrug users. Seventy-five individuals with an addiction to heroin, hashish and/or polydrug use were recruited to this study. BChE levels in the plasma were determined by Ellman's principle. SNPs were genotyped by standard procedures, followed by Sanger sequencing. Plasma BChE levels were found to be significantly higher (p ≤ 0.05) in addicts (mean ± standard error of the mean 0.031 ± 0.004μmol/L/min; 95% confidence interval [CI] 0.024-0.038) than in non-addicts (controls) (0.014 ± 0.001μmol/L/min; 95% CI 0.012-0.017). Statistical significant differences were also observed between the addicted cohorts. A statistically significant association for both SNPs (rs3495 and rs1803274) was not observed in addicted subjects tested in the dominant, recessive and allele genetic models, but trends of variations of the rs3495 risk G allele were noted. The authors conclude that BChE plays significant roles in addiction pathophysiology as increased BChE activity in blood samples obtained from the cohorts with addiction was evident. Further studies in this direction may provide novel approaches for the treatment of addiction, but studies with a larger sample size and different ethnic groups are warranted for broader conclusions to be drawn.

Assay optimisation and age-related baseline variation in biochemical markers in Lesser Black-backed gulls

Free-ranging animals are often used as bioindicators of both short- and long-term changes in ecosystem health, mainly to detect the presence and effects of contaminants. Birds, and gulls in particular, have been used as bioindicators over a broad range of marine and terrestrial ecosystems. In this study, we standardise the conditions for the use of a suite of biochemical markers in non-destructive matrices of Lesser Black-backed Gull (Larus fuscus) to facilitate future biomonitoring of marine and terrestrial contaminants. We characterized cholinesterase (ChE) in plasma and optimized assay conditions for ChE activity as a marker of neurotoxic damage. Moreover, we quantified variation in activity of ChE, lactate dehydrogenase (LDH), glutathione-S-transferase (GST) and catalase (CAT) as well as variation ranges of lipid peroxidation (LPO), in free-ranging adults and captive chicks. The main ChE form present in plasma of both adults and chicks was butyrylcholinesterase (BChE) followed by acetylcholinesterase (AChE), whose relative proportion in plasma tended to decrease with increased chick age. LPO levels and GST activity in blood cells (BCs) decreased significantly with increasing chick age, while BChE and LDH activity in plasma were not age-dependent. CAT in BCs tended to decline non-significantly in older chicks. Results of this study underscore the importance of standardising assay conditions and assessing intrinsic baseline variation in biochemical markers, before biochemical quantification. Data presented here provide a foundation for future use of BChE and LDH activity in plasma, as well as oxidative stress markers (LPO, CAT and GST) in BCs, to monitor environmental stress effects in Lesser Black-backed gulls.