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Frontiers in Pharmacology 2023Organophosphate-based chemical agents (OP), including nerve agents and certain pesticides such as paraoxon, are potent acetylcholinesterase inhibitors that cause severe...
Organophosphate-based chemical agents (OP), including nerve agents and certain pesticides such as paraoxon, are potent acetylcholinesterase inhibitors that cause severe convulsions and seizures, leading to permanent central nervous system (CNS) damage if not treated promptly. The current treatment regimen for OP poisoning is intramuscular injection of atropine sulfate with an oxime such as pralidoxime (2-PAM) to mitigate cholinergic over-activation of the somatic musculature and autonomic nervous system. This treatment does not provide protection against CNS cholinergic overactivation and therefore convulsions require additional medication. Benzodiazepines are the currently accepted treatment for OP-induced convulsions, but the convulsions become refractory to these GABA agonists and repeated dosing has diminishing effectiveness. As such, adjunct anticonvulsant treatments are needed to provide improved protection against recurrent and prolonged convulsions and the associated excitotoxic CNS damage that results from them. Previously we have shown that brief, 4-min administration of 3%-5% isoflurane in 100% oxygen has profound anticonvulsant and CNS protective effects when administered 30 min after a lethal dose of paraoxon. In this report we provide an extended time course of the effectiveness of 5% isoflurane delivered for 5 min, ranging from 60 to 180 min after a lethal dose of paraoxon in rats. We observed substantial effectiveness in preventing neuronal loss as shown by Fluoro-Jade B staining when isoflurane was administered 1 h after paraoxon, with diminishing effectiveness at 90, 120 and 180 min. magnetic resonance imaging (MRI) derived T2 and mean diffusivity (MD) values showed that 5-min isoflurane administration at a concentration of 5% prevents brain edema and tissue damage when administered 1 h after a lethal dose of paraoxon. We also observed reduced astrogliosis as shown by GFAP immunohistochemistry. Studies with continuous EEG monitoring are ongoing to demonstrate effectiveness in animal models of soman poisoning.
PubMed: 38230376
DOI: 10.3389/fphar.2023.1293280 -
Biochemia Medica Feb 2024Paraoxonase 1 (PON1) is the enzyme that removes carcinogenic radicals from lipids. The aim of the study was to investigate the differences in PON1 activity and oxidation...
INTRODUCTION
Paraoxonase 1 (PON1) is the enzyme that removes carcinogenic radicals from lipids. The aim of the study was to investigate the differences in PON1 activity and oxidation stress parameters between patients with cervical intraepithelial neoplasia (CIN) and healthy controls.
MATERIALS AND METHODS
The study included 65 women with CIN and 109 healthy women. Lipid parameters were determined on Cobas Integra 400 plus (Roche, Mannheim, Germany). Tiols and reduced glutathione (GSH) were determined spectrophotometric using Eliman reagent. Activity of PON1 was assessed with two substrates, paraoxon and phenylacetate by spectrophotometric method. Malondialdehyde (MDA) was determined by high performance liquid chromatography (Shimadzu Corporation, Kyoto, Japan). Mann-Whitney-test, t-test, χ2-test, correlation and logistic regression was used in statistical analysis. P < 0.05 was considered statistically significant.
RESULTS
The basal (P = 0.929) and NaCl-stimulated (P = 0.985) PON1 activity and activities standardised on the concentration of high-density lipoprotein (HDL; P = 0.076; P = 0.065, respectively) and apolipoprotein AI (apo AI; P = 0.444; P = 0.499, respectively) as well as PON1 phenotypes (P = 0.842) did not differ significantly between the groups. The PON1 arylesterase activity (53±19 kU/L vs. 77±17 kU/L; P < 0.001) and HDL-standardized activity (37 (28-44) kU/mmol . 43 (37-50) kU/mmol; P < 0.001) and apoAI (29±11 kU/g . 44±11 kU/g; P < 0.001) was significantly reduced in the CIN group. The concentration of the thiol groups was similar (P = 0.519), of MDA was lower (0.39 (0.27-0.55) µmol/L . 0.76 (0.57-1.15) µmol/L; P < 0.001) and of GSH was higher (112.0 (66.0-129.6) µg/mL . 53.4 (34.8-134.4) µg/mL; P < 0.001) in the CIN group.
CONCLUSION
Reduced PON1 arylesterase activity, lower MDA and higher GSH concentration were observed in CIN patients.
Topics: Humans; Female; Aryldialkylphosphatase; Carboxylic Ester Hydrolases; Oxidative Stress; Uterine Cervical Dysplasia
PubMed: 38125616
DOI: 10.11613/BM.2024.010701 -
RSC Advances Oct 2023The use of organophosphate (OPs) pesticides is widespread in agriculture and horticulture, but these chemicals can be lethal to humans, causing fatalities and deaths...
The use of organophosphate (OPs) pesticides is widespread in agriculture and horticulture, but these chemicals can be lethal to humans, causing fatalities and deaths each year. The inhibition of acetylcholinesterase (AChE) by OPs leads to the overstimulation of cholinergic receptors, ultimately resulting in respiratory arrest, seizures, and death. Although 2-pralidoxime (2-PAM) is the FDA-approved drug for treating OP poisoning, there is difficulty in blood-brain barrier permeation. To address this issue, we designed and evaluated a series of 2-PAM analogs by substituting electron-donating groups on the and/or positions of the pyridinium core using techniques. Our PCM-ONIOM2 (MP2/6-31G*:PM7//B3LYP/6-31G*:UFF) binding energy results demonstrated that 13 compounds exhibited higher binding energy than 2-PAM. The analog with phenyl and methyl groups substituted on the and positions, respectively, showed the most favorable binding characteristics, with aromatic residues in the active site (Y124, W286, F297, W338, and Y341) and the catalytic residue S203 covalently bonding with paraoxon. The results of DS-MD simulation revealed a highly favorable apical conformation of the potent analog, which has the potential to enhance reactivation of AChE. Importantly, newly designed compound demonstrated appropriate drug-likeness properties and blood-brain barrier penetration. These results provide a rational guide for developing new antidotes to treat organophosphate insecticide toxicity.
PubMed: 37928857
DOI: 10.1039/d3ra03087c -
The Journal of Pharmacology and... Jan 2024Organophosphate (OP) compounds are highly toxic and include pesticides and chemical warfare nerve agents. OP exposure inhibits the acetylcholinesterase enzyme, causing...
Organophosphate (OP) compounds are highly toxic and include pesticides and chemical warfare nerve agents. OP exposure inhibits the acetylcholinesterase enzyme, causing cholinergic overstimulation that can evolve into status epilepticus (SE) and produce lethality. Furthermore, OP-induced SE survival is associated with mood and memory dysfunction and spontaneous recurrent seizures (SRS). In male Sprague-Dawley rats, we assessed hippocampal pathology and chronic SRS following SE induced by administration of OP agents paraoxon (2 mg/kg, s.c.), diisopropyl fluorophosphate (4 mg/kg, s.c.), or O-isopropyl methylphosphonofluoridate (GB; sarin) (2 mg/kg, s.c.), immediately followed by atropine and 2-PAM. At 1-hour post-OP-induced SE onset, midazolam was administered to control SE. Approximately 6 months after OP-induced SE, SRS were evaluated using video and electroencephalography monitoring. Histopathology was conducted using hematoxylin and eosin (H&E), while silver sulfide (Timm) staining was used to assess mossy fiber sprouting (MFS). Across all the OP agents, over 60% of rats that survived OP-induced SE developed chronic SRS. H&E staining revealed a significant hippocampal neuronal loss, while Timm staining revealed extensive MFS within the inner molecular region of the dentate gyrus. This study demonstrates that OP-induced SE is associated with hippocampal neuronal loss, extensive MFS, and the development of SRS, all hallmarks of chronic epilepsy. SIGNIFICANCE STATEMENT: Models of organophosphate (OP)-induced SE offer a unique resource to identify molecular mechanisms contributing to neuropathology and the development of chronic OP morbidities. These models could allow the screening of targeted therapeutics for efficacious treatment strategies for OP toxicities.
Topics: Rats; Male; Animals; Rats, Sprague-Dawley; Mossy Fibers, Hippocampal; Organophosphates; Acetylcholinesterase; Status Epilepticus; Epilepsy; Seizures; Disease Models, Animal
PubMed: 37643794
DOI: 10.1124/jpet.123.001739 -
Bioorganic & Medicinal Chemistry Letters Nov 2023This study aimed to explore non-pyridinium oxime acetylcholinesterase (AChE) reactivators that could hold the potential to overcome the limitations of the currently...
This study aimed to explore non-pyridinium oxime acetylcholinesterase (AChE) reactivators that could hold the potential to overcome the limitations of the currently available compounds used in the clinic to treat the neurologic manifestations induced by intoxication with organophosphorus agents. Fifteen compounds with various non-pyridinium oxime moieties were evaluated for AChE activity at different concentrations, including aldoximes, ketoximes, and α-ketoaldoximes. The therapeutic potential of the oxime compounds was evaluated by assessing their ability to reactivate AChE inhibited by paraoxon. Among the tested compounds, α-Ketoaldoxime derivative 13 showed the highest reactivation (%) reaching 67 % and 60 % AChE reactivation when evaluated against OP-inhibited electric eel AChE at concentrations of 1,000 and 100 μM, respectively. Compound 13 showed a comparable reactivation ability of AChE (60 %) compared to that of pralidoxime (56 %) at concentrations of 100 μM. Molecular docking simulation of the most active compounds 12 and 13 was conducted to predict the binding mode of the reactivation of electric eel AChE. As a result, a non-pyridinium oxime moiety 13, is a potential reactivator of OP-inhibited AChE and is taken as a lead compound for the development of novel AChE reactivators with enhanced capacity to freely cross the blood-brain barrier.
Topics: Oximes; Paraoxon; Acetylcholinesterase; Cholinesterase Reactivators; Cholinesterase Inhibitors; Molecular Docking Simulation; Pyridinium Compounds; Acetamides; Organophosphorus Compounds
PubMed: 37838342
DOI: 10.1016/j.bmcl.2023.129504 -
Micromachines Aug 2023The ongoing advancement in the synthesis of new nanomaterials has accelerated the rapid development of non-enzymatic pesticide sensors based on electrochemical...
Development and Characterization of Nano-Ink from Silicon Carbide/Multi-Walled Carbon Nanotubes/Synthesized Silver Nanoparticles for Non-Enzymatic Paraoxon Residuals Detection.
The ongoing advancement in the synthesis of new nanomaterials has accelerated the rapid development of non-enzymatic pesticide sensors based on electrochemical platforms. This study aims to develop and characterize Nano-ink for applying organophosphorus pesticides using paraoxon residue detection. Multi-walled carbon nanotubes, silicon carbide, and silver nanoparticles were used to create Nano-ink using a green synthesis process in 1:1:0, 1:1:0.5, and 1:1:1 ratios, respectively. These composites were combined with chitosan of varying molecular weights, which served as a stabilizing glue to keep the Nano-ink employed in a functioning electrode stable. By using X-ray powder diffraction, Raman spectroscopy, energy dispersive X-ray spectroscopy, and a field emission scanning electron microscope, researchers were able to examine the crystallinity, element composition, and surface morphology of Nano-ink. The performance of the proposed imprinted working electrode Nano-ink was investigated using cyclic voltammetry and differential pulse voltammetry techniques. The Cyclic voltammogram of Ag NPs/chitosan (medium, 50 mg) illustrated high current responses and favorable conditions of the Nano-ink modified electrode. Under the optimized conditions, the reduction currents of paraoxon using the DPV techniques demonstrated a linear reaction ranging between 0.001 and 1.0 µg/mL (R = 0.9959) with a limit of detection of 0.0038 µg/mL and a limit of quantitation of 0.011 µg/mL. It was concluded that the fabricated Nano-ink showed good electrochemical activity for non-enzymatic paraoxon sensing.
PubMed: 37630149
DOI: 10.3390/mi14081613 -
RSC Advances Sep 2023The present study aims to design and synthesise novel uncharged aldoximes and explore their reactivation abilities, structures, descriptors, and mechanisms of action, as...
The present study aims to design and synthesise novel uncharged aldoximes and explore their reactivation abilities, structures, descriptors, and mechanisms of action, as well as assessing the interactions and stabilities in the active site of paraoxon-inhibited acetylcholinesterase enzyme using computational studies and assay. The comprehensive computational studies including quantum chemical, molecular dynamics simulations and molecular docking were conducted on paraoxon-inhibited human acetylcholinesterase to investigate the reactivation ability of the novel aldoximes and compare them with pralidoxime as a reactivator model molecule.
PubMed: 37780731
DOI: 10.1039/d3ra05658a -
Biosensors & Bioelectronics: X Dec 2023Organophosphate (OP) pesticides remain a worldwide health concern due to their acute or chronic poisoning and widespread use in agriculture around the world. There is a...
Organophosphate (OP) pesticides remain a worldwide health concern due to their acute or chronic poisoning and widespread use in agriculture around the world. There is a need for robust and field-deployable tools for onsite detection of OP pesticides in food and water. Herein, we present an integrated smartphone/resistive biosensor for simple, rapid, reagentless, and sensitive monitoring of OP pesticides in food and environmental water. The biosensor leverages the hydrolytic activity of acetylcholinesterase (AChE) to its substrate, acetylcholine (ACh), and unique transport properties of polyaniline nanofibers (PAnNFs) of chitosan/AChE/PAnNF/carbon nanotube (CNT) nanocomposite film on a gold interdigitated electrode. The principle of the sensor relies on OP inhibiting AChE, thus, reducing the rate of ACh hydrolysis and consequently decreasing the rate of protons doping the PAnNFs. Such resulted decrease in conductance of PAnNF can be used to quantify OP pesticides in a sample. A mobile app for the biosensor was developed for analyzing measurement data and displaying and sharing testing results. Under optimal conditions, the biosensor demonstrated a wide linear range (1 ppt-100 ppb) with a low detection limit (0.304 ppt) and high reproducibility (RSD <5%) for Paraoxon-Methyl (PM), a model analyte. Furthermore, the biosensor was successfully applied for analyzing PM spiked food/water samples with an average recovery rate of 98.3% and provided comparable results with liquid chromatography-mass spectrometry. As such, the nanosensing platform provides a promising tool for onsite rapid and sensitive detection of OP pesticides in food and environmental water.
PubMed: 38124900
DOI: 10.1016/j.biosx.2023.100402 -
GeoHealth Apr 2024The Multi-Threat Medical Countermeasure (MTMC) technique is crucial for developing common biochemical signaling pathways, molecular mediators, and cellular processes....
The Multi-Threat Medical Countermeasure (MTMC) technique is crucial for developing common biochemical signaling pathways, molecular mediators, and cellular processes. This study revealed that the Nod-like receptor 3 (NLRP3) inflammasome pathway may be a significant contributor to the cytotoxicity induced by various organophosphorus pesticides (OPPs). The study demonstrated that exposure to six different types of OPPs (paraoxon, dichlorvos, fenthion, dipterex, dibrom, and dimethoate) led to significant cytotoxicity in BV2 cells, which was accompanied by increased expression of NLRP3 inflammasome complexes (NLRP3, ASC, Caspase-1) and downstream inflammatory cytokines (IL-1β, IL-18), in which the order of cytotoxicity was dichlorvos > dipterex > dibrom > paraoxon > fenthion > dimethoate, based on the IC values of 274, 410, 551, 585, 2,158, and 1,527,566 μM, respectively. The findings suggest that targeting the NLRP3 inflammasome pathway could be a potential approach for developing broad-spectrum antitoxic drugs to combat multi-OPPs-induced toxicity. Moreover, inhibition of NLRP3 efficiently protected the cells against cytotoxicity induced by these six OPPs, and the expression of NLRP3, ASC, Caspase-1, IL-1β, and IL-18 decreased accordingly. The order of NLRP3 affinity for OPPs was dimethoate > paraoxon > dichlorvos > dibrom > (fenthion and dipterex) based on values of 89.8, 325, 1,460, and 2,690 μM, respectively. Furthermore, the common molecular mechanism of NLRP3-OPPs was clarified by the presence of toxicity effector groups (benzene ring, nitrogen/oxygen-containing functional group); =O, -O-, or =S (active) groups; and combination residues (Gly271, Asp272). This finding provided valuable insights into exploring the common mechanisms of multiple threats and developing effective therapeutic strategies to prevent OPPs poisoning.
PubMed: 38638206
DOI: 10.1029/2023GH000888 -
International Journal of Nanomedicine 2024Organophosphates are among the deadliest of known chemicals based on their ability to inactivate acetylcholinesterase in neuromuscular junctions and synapses of the...
A Pralidoxime Nanocomplex Formulation Targeting Transferrin Receptors for Reactivation of Brain Acetylcholinesterase After Exposure of Mice to an Anticholinesterase Organophosphate.
INTRODUCTION
Organophosphates are among the deadliest of known chemicals based on their ability to inactivate acetylcholinesterase in neuromuscular junctions and synapses of the central and peripheral nervous systems. The consequent accumulation of acetylcholine can produce severe acute toxicities and death. Oxime antidotes act by reactivating acetylcholinesterase with the only such reactivator approved for use in the United States being 2-pyridine aldoxime methyl chloride (., pralidoxime or 2-PAM). However, this compound does not cross the blood-brain barrier readily and so is limited in its ability to reactivate acetylcholinesterase in the brain.
METHODS
We have developed a novel formulation of 2-PAM by encapsulating it within a nanocomplex designed to cross the blood-brain barrier via transferrin receptor-mediated transcytosis. This nanocomplex (termed scL-2PAM) has been subjected to head-to-head comparisons with unencapsulated 2-PAM in mice exposed to paraoxon, an organophosphate with anticholinesterase activity.
RESULTS AND DISCUSSION
In mice exposed to a sublethal dose of paraoxon, scL-2PAM reduced the extent and duration of cholinergic symptoms more effectively than did unencapsulated 2-PAM. The scL-2PAM formulation was also more effective than unencapsulated 2-PAM in rescuing mice from death after exposure to otherwise-lethal levels of paraoxon. Improved survival rates in paraoxon-exposed mice were accompanied by a higher degree of reactivation of brain acetylcholinesterase.
CONCLUSION
Our data indicate that scL-2PAM is superior to the currently used form of 2-PAM in terms of both mitigating paraoxon toxicity in mice and reactivating acetylcholinesterase in their brains.
Topics: Animals; Mice; Acetylcholinesterase; Brain; Cholinesterase Inhibitors; Cholinesterase Reactivators; Organophosphates; Oximes; Paraoxon; Pralidoxime Compounds
PubMed: 38229703
DOI: 10.2147/IJN.S443498