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To investigate the associations between isocarbophos and isofenphos with impaired fasting glucose (IFG) and type 2 diabetes mellitus (T2DM), and to assess the mediation roles of inflammation cells. There were 2701 participants in the case-control study, including 896 patients with T2DM, 900 patients with IFG, 905 subjects with NGT. Plasma isocarbophos and isofenphos concentrations were measured using gas chromatography and triple quadrupole tandem mass spectrometry. Generalized linear models were used to calculate the relationships between plasma isofenphos and isocarbophos levels with inflammatory factor levels and T2DM. Inflammatory cell was used as mediators to estimate the mediating effects on the above associations. Isocarbophos and isofenphos were positively related with T2DM after adjusting for other factors. The odds ratio (95% confidence interval) (OR (95%CI)) for T2DM was 1.041 (1.015, 1.068) and for IFG was 1.066 (1.009, 1.127) per unit rise in ln-isocarbophos. The prevalence of T2DM increased by 6.4% for every 1 unit more of ln-isofenphos (OR (95% CI): 1.064 (1.041, 1.087)). Additionally, a 100% rise in ln-isocarbophos was linked to 3.3% higher ln-HOMA2IR and a 0.029 mmol/L higher glycosylated hemoglobin (HbA1c) (95% CI: 0.007, 0.051). While a 100% rise in ln-isofenphos was linked to increase in ln-HOMA2 and ln-HOMA2IR of 5.8% and 3.4%, respectively. Furthermore, white blood cell (WBC) and neutrophilic (NE) were found to be mediators in the relationship between isocarbophos and T2DM, and the corresponding proportions were 17.12% and 17.67%, respectively. Isofenphos and isocarbophos are associated with IFG and T2DM in the rural Chinese population, WBC and NE have a significant role in this relationship., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Organophosphate pesticides (OPs) are widely utilized in agricultural production, and the residues threaten public health and environmental safety due to their toxicity. Herein, a novel and simple DNA aptamer-based sensor has been fabricated for the rapid, visual, and quantitative detection of profenofos and isocarbophos. The proposed DNA aptamers with a G-quadruplex spatial structure could be recognized by SYBR Green I (SG-I), resulting in strong green fluorescence emitted by SG-I. The DNA aptamers exhibit a higher specific binding ability to target OP molecules through aromatic ring stacking, disrupting the interaction between SG-I and DNA aptamers to induce green fluorescence quenching. Meanwhile, the fluorescence wavelength of G-quadruplex fluorescence emission peaks changes, accompanied by an obvious fluorescence variation from green to blue. SG-I-modified aptasensor without any additive reference fluorescence units for use in multicolor fluorescence assay for selective monitoring of OPs was first developed. The developed aptasensor provides a favorable linear range from 0 to 200 nM, with a low detection limit of 2.48 and 3.01 nM for profenofos and isocarbophos, respectively. Moreover, it offers high selectivity and stability in real sample detection with high recoveries. Then, a self-designed portable smartphone sensing platform was successfully used for quantitative result outputs, demonstrating experience in designing a neotype sensing strategy for point-of-care pesticide monitoring.

A series of uncharged salicylaldehyde oximes were synthesized and evaluated for the reactivation of organophosphorus (OP) nerve agents simulants Diethylchlorophosphonate (DCP) & Diethylcyanophosphonate (DCNP) and pesticides (paraoxon & malaoxon) inhibited electric eel Acetylcholinesterase ( AChE). The computational software Swiss ADME and molinspiration were used to unfold the probability of drug-likeness properties of the oximes derivatives. Substituted aromatic oximes with diethylamino or bromo group with free hydroxyl group ortho to oxime moiety were found efficient to regenerate the enzymatic activity in in-vitro AChE assay. The alkylation of the ortho hydroxyl group of derivatives led to the loss of reactivation potential. The derivatives with a hydroxyl group and without oxime group and vice versa did not show significant reactivation potency against tested OP toxicants. Further, we also evaluated the reactivation potential of these selected molecules on the rat brain homogenate against different OPs inhibited ChE and found maximum reactivation potency of oxime 2e . The in-vitro results were further validated by molecular docking and dynamic studies which showed that the hydroxyl group interacted with serine amino acids in the catalytic anionic site of AChE enzyme and was stable up to 200 ns consequently providing proper orientation to oxime moiety for reactivating the OP inhibited enzyme. It has thus been proved by the structure-activity relationship of oximes derivatives that hydroxyl group ortho to oxime is essential for reactivating OP inhibited electric eel AChE. Amongst the twenty-one oximes derivatives, 2e was found to be most active in regenerating the paraoxon, malaoxon, DCP and DCNP inhibited AChE enzyme.

The traditional preparation of MXeneTi 3 C 2 is complicated. Two-dimensional MXeneTi 3 C 2 nanosheets were prepared via in-situ generation of HF etching Ti 3 AlC 2 under the microwave irradiation. Gold nanocluster doped nanosheets sol (MXene@Au) with excellent stability and strong catalysis was synthesized with no product residue reducer CO. MXene@Au nanosol can catalyze mandelic acid-HAuCl 4 to produce gold nanoparticles, which can be traced via resonance Rayleigh scattering (RRS) and surface-enhanced Raman scattering (SERS). Coupled the dual-mode nanocatalytic indicator reaction with aptamer reaction, a high sensitivity, selectivity, facile SERS/RRS dual-mode biosensnoring assay platform has been constructed for isocarbophos (ICP). The linear range of SERS was 1.0 × 10 -3 -2.5 × 10 -2  nmol/L, with a detection of limit 4.5 × 10 -5  nmol/L (S/N = 3). It has been used to detect ICP in water samples, with a good recovery (95.5-104%) and a good relative standard deviation (5.2-9.6%). This new nanocatalytic amplification biosensoring strategy can also assay other organic pesticides including malathion, glyphosate, profenofos and carbendazim. In addition, the nanocatalytic mechanism was investigated., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Bis-propargylcalix[4]arene-stabilized silver nanoparticles (BPCA-Ag NPs), as a chemosensor for detecting an isocarbophos (ICP) pesticide in an aqueous medium, are reported in this work. The nanoparticles were characterized by UV-visible spectroscopy, dynamic light scattering, zeta potential and high-resolution transmission electron microscopy techniques. It was observed that the BPCA-Ag NPs had a high selectivity for isocarbophos with a detection limit of 1.0 × 10 -6  M. According to the result of this research, the BPCA-Ag NPs were found to be useful for the colorimetric detection of isocarbophos in an aqueous medium. It provides a new method for in situ detection of isocarbophos using host-guest interaction., (© 2022. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.)

A novel label-free and enzyme-free detection strategy has been developed for the electrochemical biosensor detection of isocarbophos (ICP) using UiO-66-NH 2 and aptamer as the signal transducers. In this work, the ICP aptamers were attached to UiO-66-NH 2 through physical mixing and chemical combination methods. In the presence of ICP, the aptamers could undergo conformational change and bind to them, which prevent the electron transfer to the surface of electrode. By comparing the two conjunction approaches of aptasensors, these proposed strategies could selectively and sensitively detect ICP with a detection limit of 6 ng mL -1 (20.74 nM) and 0.9 ng mL -1 (3.11 nM). Furthermore, we have also demonstrated the capability of this strategy in the detection of ICP in real samples from vegetable and fruit extract, indicating the potential application of this strategy in food safety issues., (Copyright © 2021 Elsevier B.V. All rights reserved.)

We have previously reported that the long-term exposure of Isocarbophos, a kind of organophosphorus compounds, induces vascular dementia (VD) in rats. Studies have also shown that organophosphorus compounds have adverse effects on offsprings. Vitamin B6 is a coenzyme mainly involved in the regulation of metabolism and has been demonstrated to ameliorate VD. Sphingosine-1-phosphate (S1P), a biologically active lipid, plays a vital role in the cardiovascular system. However, whether S1P is involved in the therapeutic effects of Vitamin B6 on posterior cerebral artery injury has yet to be further answered. In the present study, we aimed to explore the potential influence of Vitamin B6 on Isocarbophos-induced posterior cerebral artery injury in offspring rats and the role of the S1P receptor in this process. We found that Vitamin B6 significantly improves the vasoconstriction function of the posterior cerebral artery in rats induced by Isocarbophos by the blood gas analysis and endothelium-dependent relaxation function assay. We further demonstrated that Vitamin B6 alleviates the Isocarbophos-induced elevation of ICAM-1, VCAM-1, IL-1, and IL-6 by using the enzyme-linked immunosorbent assay kits. By performing immunofluorescence and the western blot assay, we revealed that Vitamin B6 prevents the down-regulation of S1P in posterior cerebral artery injury. It is worth noting that Fingolimod, the S1P inhibitor, significantly inhibits the Vitamin B6-induced up-regulation of S1P in posterior cerebral artery injury. Collectively, our data indicate that Vitamin B6 may be a novel drug for the treatment of posterior cerebral artery injury and that S1P may be a drug target for its treatment., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Locust plagues are still worldwide problems. Selecting active enantiomers from current chiral insecticides is necessary for controlling locusts and mitigating the pesticide pollution in agricultural lands. Herein, two enantiomers of isocarbophos (ICP) were separated and the enantioselectivity in insecticidal activity against the pest Locusta migratoria manilensis (L. migratoria) and mechanisms were investigated. The significant difference of LD 50 between (+)-ICP (0.609 mg/kg bw) and (-)-ICP (79.412 mg/kg bw) demonstrated that (+)-ICP was a more effective enantiomer. The enantioselectivity in insecticidal activity of ICP enantiomers could be attributed to the selective affinity to acetylcholinesterase (AChE). Results of in vivo and in vitro assays suggested that AChE was more sensitive to (+)-ICP. In addition, molecular docking showed that the -CDOKER energies of (+)-ICP and (-)-ICP were 25.6652 and 24.4169, respectively, which suggested a stronger affinity between (+)-ICP and AChE. Significant selectivity also occurred in detoxifying enzymes activities (carboxylesterases (CarEs) and glutathione S-transferases (GSTs)) and related gene expressions. Suppression of detoxifying enzymes activities with (+)-ICP treatment suggested that (-)-ICP may induce the detoxifying enzyme-mediated ICP resistance. A more comprehensive understanding of the enantioselectivity of ICP is necessary for improving regulation and risk assessment of ICP., (Copyright © 2021. Published by Elsevier Inc.)

The chiral pesticide enantiomers often show selective efficacy and non-target toxicity. In this study, the enantioselective degradation characteristics of the chiral organophosphorus insecticide isocarbophos (ICP) by Cupriavidus nantongensis X1 T were investigated systematically. Strain X1 T preferentially degraded the ICP R isomer (R-ICP) over the S isomer (S-ICP). The degradation rate constant of R-ICP was 42-fold greater than S-ICP, while the former is less bioactive against pest insects but more toxic to humans than the latter. The concentration ratio of S-ICP to R-ICP determines whether S-ICP can be degraded by strain X1 T . S-ICP started to degrade only when the ratio (C S-ICP /C R-ICP ) was greater than 62. Divalent metal cations could improve the degradation ability of strain X1 T . The detected metabolites that were identified suggested a novel hydrolysis pathway, while the hydrolytic metabolites were less toxic to fish and green algae than those from P-O bond breakage. The crude enzyme degraded both R-ICP and S-ICP in a similar rate, indicating that enantioselective degradation was due to the transportation of strain X1 T . The strain X1 T also enantioselectively degraded the chiral organophosphorus insecticides isofenphos-methyl and profenofos. The enantioselective degradation characteristics of strain X1 T make it suitable for remediation of chiral organophosphorus insecticide contaminated soil and water., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Malathion is a widely used chiral phosphorus insecticide, which has a more toxic chiral metabolite malaoxon. In this work, the enantiomers of malathion and malaoxon were separated by high-performance liquid chromatography-mass/mass (HPLC-MS/MS) with chiral columns using acetonitrile/water or methanol/water as mobile phase, and the chromatographic conditions were optimized. Based on the chiral separation, the chiral residue analysis methods for the enantiomers in soil, fruit, and vegetables were set up. Two pairs of the enantiomers were better separated on CHIRALPAK IC chiral column, and baseline simultaneous separations of malathion and malaoxon enantiomers were achieved with acetonitrile/water (40/60, v/v) as mobile phase at a flow rate of 0.5 mL/min. The elution orders were -/+ for both malathion and malaoxon measured by an optical rotation detector. The chiral residue analysis in soil, fruit, and vegetables was validated by linearity, recovery, precision, limit of detection (LOD), and limit of quantification (LOQ). The LODs and LOQs for the enantiomers of malathion were 1 μg/kg and 3-5 μg/kg and 0.08 μg/kg and 0.20-0.25 μg/kg for malaoxon enantiomers. Good linear calibration curves for each enantiomer in the matrices were obtained within the concentration range of 0.02-12 mg/L. The mean recoveries of the enantiomers of malathion and malaoxon ranged from 82.26% to 109.04%, with RSDs of 0.71-8.63%.The results confirmed that this method was capable of simultaneously determining the residue of malathion and malaoxon in food and environmental matrix on an enantiomeric level., (© 2020 Wiley Periodicals LLC.)

Background: The oriental fruit fly, Bactrocera dorsalis (Hendel), is a widespread agricultural pest that has evolved resistance to many commonly used insecticides including malathion. Glutathione S-transferases (GSTs) are multifunctional enzymes that metabolize insecticides directly or indirectly. The specific mechanism used by GSTs to confer malathion resistance in B. dorsalis is unclear., Results: BdGSTd9 was identified from B. dorsalis and was expressed at twice the level in a malathion-resistant strain (MR) than in a susceptible strain (MS). By using RNAi of BdGSTd9, the toxicity of malathion against MR was increased. Protein modelling and docking of BdGSTd9 with malathion and malaoxon indicated key amino acid residues for direct binding in the active site. In vitro assays with engineered Sf9 cells overexpressing BdGSTd9 demonstrated lower cytotoxicity of malathion. High performance liquid chromatography (HPLC) analysis indicated that malathion could be broken down significantly by BdGSTd9, and it also could deplete the malathion metabolite malaoxon, which possesses a higher toxicity to B. dorsalis. Taken together, the BdGSTd9 of B. dorsalis could not only deplete malathion, but also react with malaoxon and therefore enhance malathion resistance., Conclusion: BdGSTd9 is a component of malathion resistance in B. dorsalis. It acts by depleting both malathion and malaoxon. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

An aptamer-based colorimetric-phosphorescent assay was developed for the detection of isocarbophos. The colorimetric assay relied on the aggregation of gold nanoparticles (AuNPs) caused by the competitive binding of aptamer between isocarbophos and AuNPs in the presence of a high salt concentration. The further addition of persistent luminescence nanorods (PLNRs) into the system showed the phosphorescence sensitively proportional to the concentration of isocarbophos, due to the inner filter effect between PLNRs and AuNPs. The assay showed good linearity within 50-500 μg/L and 5-160 μg/L, and limit of detection of 7.1 μg/L and 0.54 μg/L in colorimetry and phosphorescence mode, respectively. The feasibility of this approach for food analysis was demonstrated with the sensitive and selective determination of isocarbophos residues in vegetables., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A novel colorimetric sensing platform based on the peroxidase activity of hemin regulated by oligonucleotide and pesticide was reported for the ultrasensitive and selective detection of isocarbophos. Oligonucleotides can accumulate on the surface of hemin in acid condition and temporarily inhibit its catalytic activity, which results in the loss of one electron of TMB molecule and produce the blue products. With the addition of isocarbophos, the pesticide molecules can interact with oligonucleotides to form some complexes, which relieve the inhibition of ssDNA to hemin and further enhance its catalytic activity. Thus, the TMB molecules are further oxidized to lose another electron and produce the yellow product in a few minutes, which has the characteristic absorption peak at 450 nm. The color change of the sensing system is related to the amount of isocarbophos, so this method can quickly discriminate whether the target pesticide exceeds the maximal residue limit just by naked eyes. To improve the performance of sensing platform, some important parameters like buffer condition and ssDNA have been investigated, and the peroxidase activity of hemin was further studied to verify the catalytic mechanism. The proposed sensing platform has a detection limit as low as 0.6 μg/L and displays good selectivity against other competitive pesticides. Moreover, the developed sensing platform also exhibits favorable accuracy and stability, indicating that it has potential applications in the detection of pesticide residues in agricultural products. Graphical abstract A novel colorimetric sensing platform based on oligonucleotides and pesticide regulation; the peroxidase catalytic activity of hemin was firstly reported for the ultrasensitive and selective detection of isocarbophos pesticide.

The enantioselective bioactivity, toxicity, and environmental behaviors of isocarbophos (ICP) were investigated. The order of the bioactivity and toxicity was S-(+) ≥ rac > R-(-), and the difference of R-(-) and S-(+) was up to 232 times. The usage of S-(+)-ICP may efficiently reduce the usage amount of rac-ICP by 35% under the same effect, and the toxicity was not increased. Based on the toxic unit analysis, the additive effect and synergistic effect of ICP enantiomers were found in the four nontarget organisms, and R-(-)-ICP might cooperate the side-effects of S-(+)-ICP. The accumulation of rac-ICP in earthworms was enantioselective with an enantioenrichment of R-(-)-ICP, so the usage of racemic ICP might increase the exposure risk of R-(-)-ICP to earthworms. From the comprehensive results, the production of enantiomer enriched S-(+)-ICP might increase bioactivity and reduce environmental pollution, while the toxicity of S-(+)-ICP to other nontarget organisms needs to be further assessed.

The enantioselective bioactivity and acute toxicity to target and non-target model species, and environmental fate of isocarbophos (ICP) in rice cultivation were investigated systematically. Bioactivity and toxicity of S-(+)-ICP was significantly greater than R-(-)-ICP, and the difference was 2.9-101 times. Based on the toxic unit analysis, the toxic interaction of ICP enantiomers for target pests was synergistic effect, while for non-target fish was concentration addition or antagonistic effect. Rac-ICP displayed equivalent bioactivity to S-(+)-ICP under the equal dosage, but the toxicity of rac-ICP to the tested fishes reduced at least 2 times. Rac-ICP is more suitable than optically pure S-(+)-ICP for rice cultivation based on the toxicity and bioactivity results. In environmental behavior experiments, the main metabolite of ICP, isocarbophos oxon (ICPO) was detected in rice plants, water, rice and rice hull samples. S-(+)-ICP and S-(+)-ICPO were more persistent than the R-form in these matrices. The comprehensive data of ICP enantiomers in rice cultivation will improve environmental and ecological risk assessment, and using racemate may be more safe and reasonable in rice cultivation system., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Isofenphos-methyl (IFP) is a very active and persistent chiral insecticide. However, IFP has lower activity against acetylcholinesterases (AChEs). Previously, it was confirmed that phosphorothioate organophosphorus pesticides with N-alkyl (POPN) require activation by oxidative desulfuration and N-dealkylation. In this work, we demonstrated that IFP could be metabolized in human liver microsomes to isofenphos-methyl oxon (IFPO, 52.7%), isocarbophos (ICP, 14.2%) and isocarbophos oxon (ICPO, 11.2%). It was found that (R)-IFP was preferentially degraded compared to the (S)-enantiomer, and the enantiomeric fraction (EF) value reached 0.61 at 60 min. However, (S)-enantiomers of the three metabolites, were degraded preferentially, and the EF values ranged from 0.34 to 0.45. Cytochrome P450 (CYP) isoforms CYP3A4, CYP2E1, and CYP1A2 and carboxylesterase enzyme have an essential role in the enantioselective metabolism of IFP; but, the enzymes that participate in the degradation of IFP metabolites are different. The AChE inhibition bioassay indicated that ICPO is the only effective inhibitor of AChE. The covalent molecular docking has proposed that the metabolites of IFP and its analogs after N-dealkylation and oxidative desulfuration will possess the highest inhibitory activity against AChE. This study is the first to demonstrate that ICPO can be regarded as a potential biomarker for the biomonitoring of IFP and ICP exposure in humans., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Commercial malathion is a racemic mixture that contains two enantiomers, and malathion has adverse effects on mammals. However, whether these two enantiomers have different effects on animals remains unclear. In this study, we tested the effect of racemate, enantiomers, and metabolite of malathion on the metabolomics profile of HepG2 cells. HepG2 cells showed distinct metabolic profiles when treated with rac-malathion, malaoxon, R-(+)-malathion, and S-(-)-malathion, and these differences were attributed to pathways in amino acid metabolism, oxidative stress, and inflammatory response. In addition, malathion treatment caused changes in amino acid levels, antioxidant activity, and expression of inflammatory genes in HepG2 cells. S-(-)-Malathion exhibited stronger metabolic perturbation than its enantiomer and racemate, consistent with the high level of cytotoxicity of S-(-)malathion. R-(+)-Malathion treatment caused significant oxidative stress in HepG2 cells but induced a weaker disturbance in the amino acid metabolism and a pro-inflammatory response compared to S-(-)-malathion and rac-malathion. Malaoxon caused more significant perturbation on antioxidase and a stronger antiapoptosis effect than its parent malathion. Our results provide insight into the risk assessment of malathion enantiomers and metabolites. We also demonstrate that a metabolomics approach can identify the discrepancy of the toxic effects and underlying mechanisms for enantiomers and metabolites of chiral pesticides.

Although isocarbophos is a widely used insecticide, its toxicity to aquatic organisms has not been well characterized. In this study, zebrafish were exposed to isocarbophos at concentrations of 50 µg L -1 and 200 µg L -1 to assess its bioaccumulation, metabolic disruption, and oxidative stress. Metabolomics analysis based on 1 H NMR spectroscopy showed that 50 µg L -1 and 200 µg L -1 isocarbophos exposure induced increases in leucine, isoleucine, valine, and alanine compared to levels in the control. Lactate, creatine, and taurine were reduced in the 50 µg L -1 isocarbophos exposure group, and only lactate decreased in response to 200 µg L -1 isocarbophos. After zebrafish were exposed to 50 and 200 µg L -1 isocarbophos for 28 days, the activities of antioxidant enzymes (SOD, CAT, and GPx) and GSH contents decreased significantly in the liver. This result indicates that there was significant oxidative stress in the liver. Furthermore, changes in metabolite profiles significantly covaried with changes in several oxidative stress endpoints based on partial least squares regression. These results will contribute to the environmental risk assessment of isocarbophos and clarify the mechanism underlying its toxicity in zebrafish., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Malathion is an organophosphorus pesticide widely used in agricultural crops, despite its toxicity. In addition, malaoxon occurs by oxidation of malathion being more toxic. The toxic effects of malathion and malaoxon in humans include hepatoxicity, breast cancer, genetic damage and endocrine disruption. The aim of this study involved assessing the effect of malathion commercial grade on Chroococcus sp., and its potential as an alternative to the removal of this pesticide and its transformation product such as malaoxon. We evaluated the effect of malathion at different concentrations (1, 25, 50, 75 and 100 ppm) on the biomass of the cyanobacteria Chroococcus sp. grown in medium BG-11; also, we analyse its ability to degrade both malathion and malaoxon into a temperature of 28 ± 2 °C and at pH 6. The results showed that 50 ppm of malathion the cyanobacteria Chroococcus sp. reached the highest removal efficiency of malathion and malaoxon (69 and 65%, respectively); also, the growth rate of Chroococcus sp. increased without inhibiting the production of chlorophyll "a", this can be explained by the hormesis phenomenon. Therefore, we consider that the cyanobacteria Chroococcus sp. may be a good candidate for bioremediation of aquatic systems contaminated with organophosphorus pesticides such as malathion and its transformation product such as malaoxon.

Background: We have previously reported that the long-term exposure of organophosphorus induces vascular dementia (VD) in rats. As a coenzyme, vitamin B6 is mainly involved in the regulation of metabolisms. Whether vitamin B6 improves VD remains unknown., Methods: The model of VD was induced by feeding rats with isocarbophos (0.5 mg/kg per two day, 12 weeks). The blood flow of the posterior cerebral artery (PCA) in rat was assessed by transcranial Doppler (TCD). The learning and memory were evaluated by the Morris Water Maze (MWM) test., Results: Administration of vitamin B6 increased the blood flow in the right and left posterior cerebral arteries and improved the functions of learning and memory in isocarbophos-treated rats. Vitamin B6 increased the protein levels of N-methyl-D-aspartate receptor (NMDAR) 2B, postsynaptic densities (PSDs) protein 95, and calmodulin-dependent protein kinase II (CaMK-II) in the hippocampus, which were decreased by isocarbophos in rats. Morphological analysis by light microscope and electronic microscope indicated disruptions of the hippocampus caused by isocarbophos were normalized by vitamin B6. Importantly, the antagonist of NMDAR signaling by eliprodil abolished these beneficial effects produced by vitamin B6 on PCA blood flow, learning, memory, and hippocampus structure in rats, as well as the protein expression of NMDAR 2B, PSDs protein 95, and CaMK-II in the hippocampus., Conclusion: Vitamin B6 activates NMDAR signaling to prevent isocarbophos-induced VD in rats.

Organophosphorus compounds (OP) represent a class of insecticides that are used most globally. The neurotoxic effects are attributed mainly to acetylcholinesterase (AChE) enzyme inhibition, which is responsible for cholinergic manifestations in individuals acutely exposed to OP. However, AChE inhibition alone cannot account for the wide range of symptoms that were reported following OP exposures. In agreement with this, evidence shows that non-cholinergic events may be mechanistically linked to OP-induced neurotoxicity. The aim of this study was to investigate the potential occurrence of oxidative stress as a critical step in the toxicity induced by the OP malaoxon(MAL) using primary cultures of mouse cortical neurons, as well as to distinguish MAL-induced oxidative stress and cell toxicity from an action on AChE blockade. Primary cultures of mouse cortical neurons were treated with MAL (0.01; 0.1; 1; 10; or 100 µM) at varying time points (1, 3, 6, 24, 48, or 144 hr) and the following biochemical parameters determined including cell viability, AChE activity, and superoxide production. MAL significantly reduced cell viability in a concentration- and time-dependent manner. Of note, 1 µM MAL significantly inhibited (approximately 75%) AChE activity after 48 hr incubation. Pralidoxime (PRAL) (600 µM), a classical AChE reactivator, significantly protected against MAL-induced AChE blockade; however, PRAL did not affect MAL-mediated fall in cellular viability, indicating that AChE inhibition is not necessarily correlated with insecticide-induced decrease in cell survival. MAL-induced diminished cell viability was preceded by a significant increase in superoxide anion production. The antioxidant agent ascorbic acid (AA) (200 µM), which significantly protected against MAL-induced superoxide anion production, did not alter MAL-induced AChE inhibition and significantly prevented insecticide-mediated fall in cell survival. Data show that increased superoxide anion production is an event that precedes MAL-induced cell toxicity in primary cultures of mouse cortical neurons. Based on the preventative effects of AA against MAL-mediated superoxide anion production and reduced cell viability, evidence indicates that oxidative stress represents an important step mediating MAL-induced toxicity in neurons and that AChE inhibition is not necessarily correlated with lowered cell survival noted in insecticide-exposed cells.

The behavior and fate of 3 pesticides (triadimefon, malathion, and dichlorvos) and the main metabolites (triadimenol and malaoxon) during barley storage or beer processing were assessed using a pilot-plant equipment. The residues of all products were determined using liquid chromatography coupled with tandem mass spectrometry. Field investigation of the dissipation rate kinetics for triadimefon and malathion during storage indicated that their half-life was twice as high when 5 times the recommended dosage was used. Milling had little effect on the removal of dichlorvos and malathion residues, whereas these were substantially removed when the spent grains were filtered after mashing. The calculated processing factors were all <1, indicating the residual ratios of dichlorvos and malathion were reduced during the entire process. In conclusion, storage and processing considerably reduced pesticide residue levels in barley and beer; however, greater focus needs to be paid to the toxicity of their metabolites in commercial by-products., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Vascular dementia, being the most severe form of vascular cognitive impairment (VCI), is caused by cerebrovascular disease. Whether organophosphorus causes VCI remains unknown. Isocarbophos (0.5 mg/kg per 2 days) was intragastrically administrated to rats for 16 weeks. The structure and function of cerebral arteries were assayed. The learning and memory were evaluated by serial tests of step-down, step-through and morris water maze. Long-term administration of isocarbophos reduced the hippocampal acetylcholinesterase (AChE) activity and acetylcholine (ACh) content but did not alter the plasma AChE activity, and significantly damaged the functions of learning and memory. Moreover, isocarbophos remarkably induced endothelial dysfunction in the middle cerebral artery and the expressions of ICAM-1 and VCAM-1 in the posterior cerebral artery. Morphological analysis by light microscopy and electron microscopy indicated disruptions of the hippocampus and vascular wall in the cerebral arteries from isocarbophos-treated rats. Treatment of isocarbophos injured primary neuronal and astroglial cells isolated from rats. Correlation analysis demonstrated that there was a high correlation between vascular function of cerebral artery and hippocampal AChE activity or ACh content in rats. In conclusion, chronic administration of isocarbophos induces impairments of memory and learning, which is possibly related to cerebral vascular dysfunction., (© 2016 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

New acetylcholinesterase (AChE) biosensor based on unsubstituted pillar[5]arene (P[5]A) as electron mediator was developed and successfully used for highly sensitive detection of organophosphate and carbamate pesticides. The AChE from electric eel was immobilized by carbodiimide binding on carbon black (CB) placed on glassy carbon electrode. The working potential of 200mV was obtained in chronoamperometric mode with the measurement time of 180 s providing best inter-biosensors precision of the results. The AChE biosensor developed made it possible to detect 1×10(-11)-1×10(-6) M of malaoxon, 1×10(-8)-7×10(-6) M of methyl-paraoxon, 1×10(-10)-2×10(-6) M of carbofuran and 7×10(-9)-1×10(-5) M of aldicarb with 10 min incubation. The limits of detection were 4×10(-12), 5×10(-9), 2×10(-11) and 6×10(-10) M, respectively. The AChE biosensor was tested in the analysis of pesticide residuals in spiked samples of peanut and beetroot. The protecting effect of P[5]A derivative bearing quaternary ammonia groups on malaoxon inhibition was shown., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Organophosphorus (OP) pesticides are known to induce pulmonary toxicity in both humans and experimental animals. To elucidate the mechanism of OP-induced cytotoxicity, we examined the effects of parathion and malathion and their respective metabolites, paraoxon and malaoxon, on primary cultured human large and small airway cells. Exposure to paraoxon and malaoxon produced a dose-dependent increase in cytotoxicity following a 24-hour exposure, while treatment with parathion or malathion produced no effects at clinically relevant concentrations. Exposure to paraoxon-induced caspase activation, but malaoxon failed to induce this response. Since caspases have a major role in the regulation of apoptosis and cell death, we evaluated OP-induced cell death in the presence of a caspase inhibitor. Pharmacological caspase inhibition protected against paraoxon-induced cell death but not malaoxon-induced cell death. These data suggest that caspase activation is a key signaling element in paraoxon-induced cell death, but not malaoxon-induced cellular death in the pulmonary epithelium., (© The Author(s) 2015.)

The benthic fauna is of great importance to assess the environmental fate of contaminations in aquatic ecosystem. In this study, tubificids were exposed to both laboratory-contaminated aqueous phases and spiked sediment to study the bioaccumulation of isocarbophos (ICP). Two types of spiked sediments were used in the spiked sediment experiment. During the exposure period, an enantioselective bioaccumulation was found in spiked water treatment, with concentrations of the (-)-ICP higher than that of the (+)-ICP, but no enantioselectivity was detected in the spiked sediment treatments. However, different bioaccumulation patterns were observed in the two spiked sediment treatments. Results showed that for spiked forest field sediment (FF sediment) incubation, bioaccumulation was governed by the concentrations in soil. Whereas ICP was bioaccumulated dominantly from overlying water in spiked Chagan Lake sediment (CG sediment) test. The dissipation rates were proved different in the two sediments and ICP dissipated much faster in CG sediment than that in FF sediment. Significant difference in ICP's half-life was also observed between worm-present and worm-free treatments in FF sediment. The detections of concentrations in overlying water indicated that much more ICP diffused to aquatic phase with the present of tubificids., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Exposure to pesticides is suspected to cause human health problems. Our study aimed to evaluate preventive effects of caffeic acid (3,4-dihydroxycinnamic acid) in the hypothalamus against malathion-induced neuropeptides gene expression alterations. Malathion at 100 mg/kg was administered intragastrically to rats alone or in combination with caffeic acid at 100 mg/kg during 4 weeks. A molecular expression of hypothalamic neuropeptides and plasmatic cholinesterase activity was investigated. Furthermore, we used in silico analysis, known as computational docking, to highlight the nature of acetylcholinesterase-malathion/caffeic acid interactions. Our findings showed differences in the responses and indicate that caffeic acid reversed malathion-induced decrease in corticotropin-releasing hormone mRNA but not brain-derived neurotrophic factor which presented an increased tendency. We suggest that caffeic acid can interact with acetylcholinesterase as the primary target of organophosphorus compounds. Results predict that caffeic acid can block partly the acetylcholinesterase gorge entrance via π-π stacking interaction with Tyr 124 and Trp 286 residues of the peripheral site leading to its stricture. Under this condition, we suggested that acetylcholine trafficking toward the catalytic site is ameliorated compared to malaoxon according to their sizes.

New acetylcholinesterase (AChE) sensor based on Ag nanoparticles decorated with macrocyclic ligand has been developed and successfully used for highly sensitive detection of organophosphate and carbamate pesticides. AChE was immobilized by carbodiimide binding on carbon black (CB) layer deposited on a glassy carbon electrode. The addition of Ag nanoparticles decreased the working potential of the biosensor from 350 to 50 mV. The AChE sensor made it possible to detect 0.4 nM-0.2 μM of malaoxon, 0.2 nM-0.2 μM of paraoxon, 0.2 nM-2.0 μM of carbofuran and 10 nM-0.20 μM of aldicarb (limits of detection 0.1, 0.05, 0.1 and 10 nM, respectively) with 10 min incubation. The AChE sensor was tested for the detection of residual amounts of pesticides in spiked samples of peanut and grape juice. The protecting effect of new macrocyclic compounds bearing quaternary ammonia fragments was shown on the example of malaoxon inhibition., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Organophosphorus pesticides are the most widely used pesticides in modern agricultural systems to ensure good harvests. Isocarbophos (ICP), with a potent acetylcholinesterase inhibitory effect is widely utilized to control a variety of leaf-eating and soil insects. However, the characteristics of the bioactivation and detoxification of ICP in humans remain unclear. In this study, the oxidative metabolism, esterase hydrolysis, and chiral inversion of ICP in human liver microsomes (HLMs) were investigated with the aid of a stereoselective LC/MS/MS method. The depletion of ICP in HLMs was faster in the absence of carboxylesterase inhibitor (BNPP) than in the presence of NADPH and BNPP, with t1/2 of 5.2 and 90 min, respectively. Carboxylesterase was found to be responsible for the hydrolysis of ICP, the major metabolic pathway. CYP3A4, CYP1A2, CYP2D6, CYP2C9, and CYP2C19 were all involved in the secondary metabolism pathway of desulfuration of ICP. Flavin-containing monooxygenase (FMO) did not contribute to the clearance of ICP. The hydrolysis and desulfuration of (±)ICP, (+)ICP, and (-)ICP in HLMs follow Michaelis-Menten kinetics. Individual enantiomers of ICP and its oxidative desulfuration metabolite isocarbophos oxon (ICPO) were found to be inhibitors of acetylcholinesterases at different extents. For example, (±)ICPO is more potent than ICP (IC50 0.031μM vs. 192μM), whereas (+)ICPO is more potent than (-)ICPO (IC50 0.017μM vs. 1.55μM). Given the finding of rapid hydrolysis of ICP and low abundance of oxidative metabolites presence in human liver, the current study highlights that human liver has a greater capacity for detoxification of ICP., (© The Author 2014. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

A simple one-step solvothermal method was applied for the preparation of magnetite/reduced graphene oxide (MRGO), and the synthetic nanocomposites with a magnetic particle size of ∼8nm were used as an adsorbent for magnetic solid phase extraction of isocarbophos (ICP) in different sample matrices prior to gas chromatography (GC) detection. The identity of the nanomaterial was confirmed using Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy and transmission electron microscopy. It was shown that Fe3O4 nanoparticles with a uniform size were homogeneously anchored on RGO nanosheets. Increased oxidation degrees of graphite oxide, big particle sizes and large loading amounts of Fe3O4 on the surface of RGO led to a decrease of adsorption capacity of MRGO to ICP. The adsorption behavior of this adsorbent was better fitted by the pseudo-second-order kinetic model. Several parameters affecting the extraction efficiency were investigated and optimized, including adsorbent dosage, extraction time, ionic strength and desorption conditions. And then, a rapid and effective method based on MRGO combined with GC was developed for the determination of ICP in aqueous samples. A linear range from 0.05 to 50ngmL(-1) was obtained with a high correlation coefficient (R(2)) of 0.9995, and the limit of detection was found to be 0.0044ngmL(-1). This method was successfully applied to the analysis of ICP in five kinds of samples, including apple, rice, lake water, cowpea and cabbage. The recoveries in different sample matrices were in the range from 81.00% to 108.51% with relative standard deviations less than 9.72%. It can be concluded that the proposed analytical method is highly-efficient, sensitive, precise, accurate and practicable., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Aphis gossypii (Glover) has been found to possess multiple mutations in the acetylcholinesterase (AChE) gene (Ace) that might involve target site insensitivity. In vitro functional expression of AChEs reveals that the resistant Ace1 (Ace1R) and Ace2 (Ace2R) were significantly less inhibited by eserine, omethoate, and malaoxon than the susceptible Ace1 (Ace1S) and Ace2 (Ace2S). Furthermore, in both the mutant and susceptible AChEs, Ace2 was significantly less sensitive to eserine, omethoate, and malaoxon than Ace1. These results suggested that both the mutant Ace1 and Ace2 were responsible for omethoate resistance, while the mutant Ace2 played a major role in insecticide resistance. The DNA copy number and transcription level of Ace2 were 1.52- and 1.88-fold higher in the ORR strain than in the OSS strain. Furthermore, the DNA copy number and transcription level of Ace2 were significantly higher than that of Ace1 in either OSS or ORR strains, demonstrating the involvement of Ace2 gene duplication in resistance. Thus, the authors conclude that omethoate resistance in cotton aphids appears to have evolved through a combination of multiple mutations and extensive Ace2R gene duplication., (Copyright © 2012 Wiley Periodicals, Inc.)

Context: Numerous studies demonstrated a limited efficacy of clinically used oximes in case of poisoning by various organophosphorus compounds. A broad spectrum oxime antidote covering all organophosphorus nerve agents and pesticides is still missing and effective (bio-)scavengers have not yet been marketed., Objective: The interactions of the available and clinically approved hydroxyethyl starch, dextran and lipid emulsions with organophosphorus nerve agents and pesticides were investigated in order to provide an in vitro base for the evaluation of these compounds in human organophosphorus poisoning., Materials and Methods: The degradation kinetics of organophosphorus compounds by the glucose derivatives and lipid emulsions were investigated with an acetylcholinesterase inhibition assay., Results: The incubation of organophosphorus compounds with TRIS-Ca(2+) buffer resulted in a time-dependent degradation of the nerve agents with half-lives of 42 min for cyclosarin, 49 min for sarin, 99 min for tabun, 107 min for soman 19 h for malaoxon and 54 h for VX. In contrast, incubation with all tested compounds resulted in a stabilisation of the organophosphorus compounds., Discussion: Our results suggest that binding of lipophilic organophosphorus compounds could result in a reduced spontaneous and enzyme-induced degradation of the toxic compounds., Conclusion: High dose lipid emulsions and glucose derivatives stabilised organophosphorus compounds in vitro.

Objective: Effects of neurophathologic changes and expression of Glu and 60 nNOS were observed in acute isocarbophos and phoxim poisoning in mice., Methods: KM male mice were randomly divided into three groups, which were control, non-treated and Huperzine A (HupA)-treated groups. The control group was given tween-80. Nontreated group was given isocarbophos (14.7 mg/kg) or phoxim (1702 mg/kg). HupA-treated group was given HupA 2h before phoxim or isocarbophos. Twenty-four hours after exposure, the whole brain was removed and adjacent coronal sections was obtained. One part of sections were stained with toluidine blue. The part of sections were used to assessed the expression of Glu and nNOS in the cortex and hippocampal of brain by immunohistochemistry., Results: Compared to control group, non-treated group was observed nissal body nembers reduced and dyeing light. The animals of HupA protective group were observed nissal body nembers reduced, but the lesional degree was lighter obviously than non-treated group. The statistically reduced of the expression of Glu (P<0.01), the elevation of nNOS (P<0.01), after Isocarbophos intoxication were observed. Compared to non-treated group, the significant elevation of Glu (P<0.01) and reduced of nNOS (P<0.01) was observed on HupA-treated groups. Whereas for phoxim treatment, no changes were observed., Conclusion: HupA have protective effect against glutamatergic systems disorder caused by Isocarbophos poisoning. Administration of HupA have no effects of the neurotransmitter changes induces by acute poisoning of phoxim. It is different for the toxic effect mechanism of the two organophosphate.

Malathion is an organophosphorous (OP) insecticide widely used for crop protection. Its degradates, malathiondiacid (MDA), malathion monoacid (MMA), dimethylphosphate (DMP), dimethylthiophosphate (DMTP) and dimethyldithiophosphate (DMDTP) are formed in strawberries and other produce. These same chemical biomarkers are measured in urine in human studies as quantitative measures of exposure. The excretion of malathion and its common biomarkers including MDA, MMA, DMP, DMTP and DMDTP at equal molar doses (73 μmol/kg b.w.) was studied following oral dosing of female Holtzmann rats (240-300 g). Following MDA administration, 36.3±5.4% was recovered as MDA, 0.05±0.02% as DMP, 5.5±0.3% as DMTP, 3.8±2.9% as DMDTP (mole percent), and totally 45.6±7.0% of administered dose in urine after 120 h (over 94% in the first 24h). Following DMTP administration, 8.3±7.7% was recovered as DMP, 46.6±16.5% as DMTP, and totally 55.0±10.3% of administered dose in urine after 120 h (over 92% in the first 24h). Similar results were obtained with other malathion biomarkers. Preformed biomarkers of malathion and other OP insecticides when ingested in produce are readily absorbed and excreted. Low level human dietary and non-occupational urine biomonitoring studies will be confounded by preformed pesticide biomarkers used to infer potential human pesticide exposure. This has profound implications for epidemiology studies where subject's biomarker excretion is used as a surrogate for OP exposures that cannot be related to a particular insecticide residue., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Chiral separation mechanism determination and absolute configuration assignment are fundamental to the development of chiral stationary phases (CSPs) and the evaluation of both the enantioselective bioactivity and fate of chiral compounds. This work investigated the process of chiral separation and the assignment of the absolute configurations of malathion, malaoxon, and isomalathion using chiral high performance liquid chromatography (HPLC) coupled with chiral detector-binding energy computations. Hydrogen bonding was found to be a very important factor in the chiral separation of isomalathion on Chiralpak AD, although it did not exhibit a significant effect on the chiral separation of malathion and malaoxon on Chiralcel OJ. Based on the sign of a chiral detector, the relationships between the cotton effect, optical dispersion and absolute configuration were established for individual enantiomers of malathion, malaoxon, and isomalathion. The elution orders of the enantiomers of malathion and malaoxon on Chiralcel OJ and the stereoisomers of isomalathion on Chiralpak AD predicted by binding energy computations were found to coincide precisely with those observed in the chiral separation experiments. The result suggests that binding energy computations can be used to assign the absolute configuration of the enantiomers of chiral compounds eluted on CSPs., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A sensitive flow injection-chemiluminescence (FI-CL) method for the determination of isocarbophos (ICP) residue on tangerines and oranges was proposed. It was found that the CL intensity from luminol-albumin CL reaction could be obviously quenched in the presence of ICP and the decrease in CL intensity was proportional to the logarithm of ICP concentrations ranging from 1.0 to 1000 pmol L(-1), giving the limit of detection of 0.3 pmol L(-1) (3σ). The proposed procedure was successfully applied to the determination of ICP residue on tangerines and oranges with recoveries varying from 92.0 to 111.0% and RSDs less than 5.0%. The possible CL mechanism of luminol-albumin-ICP reaction was discussed, and ICP to albumin's binding constant (K(D)=1.00 × 10(6) L mol(-1)) and the number of binding sites (n=1.00) were given by the homemade FI-CL model., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

An enantioselective method is presented for the determination of isocarbophos in soil by liquid chromatography coupled with tandem mass spectrometry. The pesticide residues in soil samples were extracted with acetonitrile, and complete enantioseparation was obtained on an amylose tris(3,5-dimethylphenylcarbamate) chiral column using acetonitrile/2 mM ammonium acetate solution containing 0.1% formic acid (60:40, v/v) as the mobile phase. The absolute configuration of isocarbophos enantiomers was determined by the combination of experimental and calculated electronic circular dichroism spectra. The method was utilized to investigate the degradation of isocarbophos in soils (Changchun, Hangzhou, and Zhengzhou) under sterilized or native conditions. Isocarbophos enantiomers were configurationally stable in the selected soils, and the pesticide degradation was not enantioselective in the sterilized condition. The degradation behavior of rac-isocarbophos was different under native conditions, with no enantioselectivity in the Changchun soil and with the S-(+)-isocarbophos enriched in the Hangzhou and Zhengzhou soils.

Among known pesticide groups, organophosphates (OPs) have grasped attention due to their hazardous nature and their applications as pesticides and chemical weapons. This work presents the development of cost-effective column based biosensor for detoxification of OPs in water and milk. Enzyme phosphotriesterase (PTE) was immobilized on an activated Sepharose 4B via covalent coupling using an Omnifit glass column. Three different OPs, ethyl paraoxon (EPOx), malaoxon (MAO) and chlorpyriphos-oxon (CPO) were spiked in water and milk to test the detoxification of OPs. Mixtures of these pesticides were also tested to check the cumulative detoxification in the real samples. The efficiency of detoxification was evaluated using a highly sensitive acetylcholinesterase (AChE) B394 biosensor based flow system. The column conditions were optimized for the detoxification studied. The method was shown to be promising when we tested real milk samples spiked with OPs. Detoxification obtained in milk was up to 86% whereas in water, 100% detoxification was obtained., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Background: Bruchid beetles, Callosobruchus species, are serious pests of economically important grain legumes; their activity in stores is often controlled by the use of synthetic insecticides. Esterases are known to be involved in insecticide resistance in insects. However, there is a dearth of information on esterase activity in the genus Callosobruchus. In this study, the effect of species, geographical strain and food type on the variation in acetylcholinesterase (AChE) activity and its inhibition by malaoxon (malathion metabolite) was investigated using an in vitro spectrophotometric method., Result: AChE activity varied significantly among species and strains and also among legume type used for rearing them. Generally, irrespective of species, strain or food type, the higher the AChE activity of a population, the higher is its inhibition by malaoxon. C. chinensis had the highest AChE activity of the species studied, and in the presence of malaoxon it had the lowest remaining AChE activity, while C. rhodesianus retained the highest activity., Conclusion: A first-hand knowledge of AChE activity in regional Callosobruchus in line with the prevailing food types should be of utmost importance to grain legume breeders, researchers on plant materials for bruchid control and pesticide manufacturer/applicators for a robust integrated management of these bruchids., (Copyright © 2012 Society of Chemical Industry.)

An inexpensive and rapid bioassay using adult red flour beetles was developed for use in assessing the decontamination of environments containing organophosphates and related chemicals. A decontamination protocol was developed which demonstrated that 2 to 3 applications of 5% bleach solution were required to obtain nearly complete decontamination of malathion. The bioassay was also used to screen common household cleaners as potential decontaminating agents, but only 5% bleach was effective at improving survival of insects on steel plates treated with 25% malathion. A toxic degradation product (malaoxon) was detected using gas chromatography/mass spectrophotometry; this toxin affected the decontamination efficacy and resulted in continued toxicity to the beetles until subsequent decontaminations. The bioassay provides evidence to support the use of red flour beetles as a sensitive, less expensive method for determining safety levels of environments contaminated with malathion and other toxins, and may have application in the study of chemical warfare agents.

Isocarbophos (ICP) is a phosphorous pesticide with high toxicity. It has been detected in several kinds of food and therefore can enter human body. In this paper, spectroscopic approaches including three-dimensional fluorescence (3D-FL) spectroscopy, UV-visible absorption spectroscopy and circular dichroism (CD) spectroscopy were employed to explore the binding of ICP to bovine serum albumin (BSA) at simulated physiological conditions. It was found that the fluorescence quenching of BSA was caused by the formation of ICP-BSA complex at ground state and belonged to static quenching mechanism. The binding constants, the number of binding sites, enthalpy change (ΔH(θ)), Gibbs free energy change (ΔG(θ)) and entropy change (ΔS(θ)) were calculated at four different temperatures according to Scatchard model and thermodynamic equations. To identify the binding location, fluorescence probe techniques were used. The results showed that warfarin, an acknowledged site marker for BSA, could be partially replaced by ICP when ICP was added to warfarin-BSA systems, which demonstrated that ICP primarily bound on Sudlow's site I in domain IIA of BSA molecule. The distance r (3.06 nm) between donor (Trp-212) and acceptor (ICP) was obtained based on Förster's non-radiation fluorescence resonance energy transfer (FRET) theory. Furthermore, the CD spectral results indicated that the secondary structure of BSA was changed in presence of ICP. The study is helpful to evaluating the toxicology of ICP and understanding its effects on the function of protein during the blood transportation process., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Chiral compounds usually behave enantioselectively in phyto-biochemical processes. Isocarbophos (ICP) is a chiral pesticide that is widely used. To evaluate the toxicological response of ICP and its enantiomers to Scenedesmus obliquus, algal growth, total chlorophyll, total soluble protein, and the superoxide anion radicals (O(2)(•-)) were investigated. The microalgae were treated with ICP and its enantiomers at 0.01-10 mg/l for 96 h. The growth of S. obliquus was stimulated at low levels of ICP and its enantiomers (0.01-1 mg/l), but all were inhibited at high concentrations (10 mg/l). The total soluble protein content and total chlorophyll content of the tested green alga S. obliquus gradually increased, depending on the growth of algal cells in the medium. Meanwhile, the content of O(2)(•-) was decreased. Interestingly, the cell number and content of the chlorophylls and protein decreased with increasing levels of concentration, whereas O(2)(•-) increased. Our results indicated that enantioselectivity was observed in the dose-response of ICP and its enantiomers in S. obliquus. The high O(2)(•-) level might lead to the death of S. obliquus. The stimulation of growth suggests a regulatory mechanism that is related to the capability of the algae to adapt to the O(2)(•-)., (Copyright © 2012 Wiley Periodicals, Inc.)

This study examined the effects of malathion and lead on transient receptor potential canonical channel TRPC1/TRPC4 channels in rat brain endothelial cells as a mechanism to explain previously noted blood-brain barrier (BBB) permeability induced by these compounds. Lead, malathion, malaoxon and combinations of these were assessed for protein levels and gene expression of TRPC1/C4 at 2, 4, 8, 16, and 24 hours after exposure. Changes in intracellular free calcium dynamics were also assessed. Compounds increased TRPC1 and TRPC4 protein levels as well as gene expression within 4 hours after exposure. Basal levels of intracellular free calcium were also elevated. Increases in gene and protein expression may be associated with an increase in the numbers of TRP channels, and the increases in intracellular calcium may be associated with activation of such channels. Therefore, upregulation and activation of the TRPC1/TRPC4 may be a mechanism by which these neurotoxicants affect BBB permeability.

An axenic bacterial strain, Lysinibacillus sp. KB1, was isolated from malathion-contaminated soil. It tolerated malathion up to 0.15 % and, under aerobic conditions, utilized it as sole carbon source. 20 % malathion and 47 % malaoxon were degraded out of the initially provided malathion. Two metabolites, mal-monocarboxylic acid and mal-dicarboxylic acid, were detected within 7 days at 30 °C. Esterase activity of the strain was 240 ± 2.5 U/ml after 7 days of growth. Sterilized soil mixed with malathion showed rapid degradation of malathion when inoculated with strain KB1 as compared to the uninoculated soil.

This work presents the development of bioassays and biosensors for the detection of insecticides widely used in the treatment of olive trees. The systems are based on the covalent immobilisation of acetylcholinesterase on magnetic microbeads using either colorimetry or amperometry as detection technique. The magnetic beads were immobilised on screen-printed electrodes or microtitration plates and tested using standard solutions and real samples. The developed devices showed good analytical performances with limits of detection much lower than the maximum residue limit tolerated by international regulations, as well as a good reproducibility and stability.

Pyridostigmine bromide (PB) has been used to protect soldiers from the toxic effects of soman, a chemical warfare agent. Recent research shows that pyridostigmine bromide protects a significant percentage of acetylcholinesterase in isolated human intercostal muscle. Findings presented here indicate that red blood cell acetylcholinesterase is similarly protected by pyridostigmine bromide from the action of diisopropyl fluorophosphate and several organophosphate pesticides including chlorpyrifos-oxon, diazinon-oxon, and paraoxon, but not malaoxon, using the bovine red blood cell as a subject. These findings suggest that pretreatment with PB may protect growers, farmworkers, first responders, and the public, in general, from the effects of selected pesticides., (Copyright © 2011 Wiley Periodicals, Inc.)

Purpose: In this study, we determined normal serum butyrylcholinesterase (BChE) and carboxylesterase (CbE) activities in Tupinambis merianae in order to obtain reference values for organophosphorus pesticide monitoring., Methods: Forty-two T. merianae individuals were grouped by sex and size to identify potential differences in their enzyme levels to allow for proper representation of normal values for females, males, juveniles, and hatchlings. Mean CbE was determined using two model substrates: alpha-naphtylacetate (α-NA) and p-nitrophenyl valerate (4-NPV). BChE and CbE sensitivity to malaoxon (Mx) was also evaluated as well as the possibility of BChE reactivation with pyridine-2-aldoxime methochloride (2-PAM)., Results: Mean adult females' BChE was significantly higher than adult males, juveniles, and hatchlings. No significant differences were found between groups regarding CbE. CbE (4-NPV) activity showed slightly negative correlation with lizard snout-vent length, while BChE and CbE (α-NA) showed no correlation with body size. Apparent IC(50) values for BChE and CbE (α-NA) suggested different sensitivities among groups. CbE (4-NPV) could not be inhibited. All Mx-inhibited groups treated with 2-PAM in a final concentration of 2.8 mM showed clear signs of reactivation., Conclusions: In conclusion, the results demonstrate that (1) plasma esterase activity did not vary with age and sex, except for BChE activity, and (2) because biological and environmental variables could be confounding factors in the response of plasma cholinesterases, complementary biomarkers like CbE inhibition and oxime-induced reactivation of esterases are strongly recommended.

Acetylcholinesterase (AChE), encoded by the Ace gene, is the primary target of organophosphates (OPs) and carbamates (CBs) in insects. Ace mutations have been identified in OP and CB resistant strains of Musca domestica. In this study, the Ace gene was partially amplified and sequenced at amino acid positions 260, 342, and 407 to determine the frequencies of these mutations in housefly samples collected from farms and garbage disposal sites of 16 provinces in the Aegean and Mediterranean regions of Turkey. In addition, the percent remaining AChE activities in these samples were assayed by using three OPs (malaoxon, paraoxon, and dichlorvos) and one CB (carbaryl) compound as inhibitors. In all the analyzed samples, 13 different combinations at the three amino acid positions were identified and the L/V260-A/G342-F/Y407 combination was found in the highest frequency. No susceptible individual was detected. The highest mean percent remaining AChE activities were detected in the individuals having the L260-A/G342-F/Y407 genotype when malaoxon and paraoxon were used as inhibitors and in the individuals with the L260-A342-F/Y407 combination when dichlorvos and carbaryl were used as inhibitors. The obtained data were heterogeneous and there was no exact correlation between the molecular genetic background and the resistance phenotypes of the flies. The findings of this study at the molecular and biochemical levels indicate the presence of significant control problems in the field., (© 2011 The Society for Vector Ecology.)

A simple and reliable technique has been developed for the construction of an amperometric acetylcholinesterase biosensor based on screen-printed carbon electrodes. For the first time, one-step modification using single-walled carbon nanotubes and Co phtalocyanine has been proposed to decrease the working potential and to increase the signal of thiocholine oxidation. The biosensor developed made it possible to detect 5-50 ppb of paraoxon and 2-50 ppb of malaoxon with detection limits of 3 and 2 ppb, respectively (incubation 15 min). The biosensor showed high reproducibility when measurements of the substrate and inhibitor were performed (R.S.D. about 1% and 2.5%, respectively). The reliability of the inhibition measurements was confirmed by testing spiked samples of sparkling and tape waters., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The simultaneous use of the repellent DEET, pyridostigmine, and organophosphorus pesticides has been assumed as a potential cause for the Gulf War Illness and combinations have been tested in different animal models. However, human in vitro data on interactions of DEET with other compounds are scarce and provoked the present in vitro study scrutinizing the interactions of DEET, pyridostigmine and pesticides with human acetylcholinesterase (hAChE) and butyrylcholinesterase (hBChE). DEET showed to be a weak and reversible inhibitor of hAChE and hBChE. The IC(50) of DEET was calculated to be 21.7mM DEET for hAChE and 3.2mM DEET for hBChE. The determination of the inhibition kinetics of pyridostigmine, malaoxon and chlorpyrifos oxon with hAChE in the presence of 5mM DEET resulted in a moderate reduction of the inhibition rate constant k(i). The decarbamoylation velocity of pyridostigmine-inhibited hAChE was not affected by DEET. In conclusion, the in vitro investigation of interactions between human cholinesterases, DEET, pyridostigmine, malaoxon and chlorpyrifos oxon showed a weak inhibition of hAChE and hBChE by DEET. The inhibitory potency of the tested cholinesterase inhibitors was not enhanced by DEET and it did not affect the regeneration velocity of pyridostigmine-inhibited AChE. Hence, this in vitro study does not give any evidence of a synergistic effect of the tested compounds on human cholinesterases., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)