Recognition mechanism and sequence optimization of organophosphorus pesticides aptamers for better monitoring contaminations in food

Aptamers as a kind of biological recognition element have shown great potential in monitoring and the rapid quantification of organophosphorus pesticides(OPPs). However, molecules of OPPs are structurally similar and original aptamers selected by systematic evolution of ligands by exponential enrichment are usually long-chain bases, which hamper the further application under OPPs-aptamer recognition. The aim of the research was to develop a new strategy to design oligonucleotide sequences for binding OPPs by combination of experimental and molecular modeling methods. 3D models of aptamers binding OPPs were constructed, and binding energy and the most probable binding site for the OPPs were then determined by molecular docking, and the binding sites were further confirmed by the results of 2-AP replaced experiments. Based on the docking results, a new aptamer for detection 4 representative OPPs with only 29 bases was designed by reasonable truncation and mutation of the reported aptamer(named S4-29). The interaction between this new aptamer and OPPs were analyzed by molecular docking, microscale thermophoresis, circular dichroism and fluorometric analysis. The results revealed that the new aptamer exhibit more superior recognition performance to OPPs, which can be promote the monitoring ability of OPPs contaminations in food.

Uncertainty Evaluation of Preparation of Organophosphorus Pesticide Mixed Standard Solution

The uncertainty of standard solution plays an important role in detection of pesticide residues. It may affect the accuracy of detection results. In this study, the 14 organophosphorus pesticides mixed standard solution was used as the material to analyze all the influencing factors for the preparation of mixed standard solution with uncertainty as the only judging index. The preparation uncertainty of mixed standard solution was calculated with the top-down calculation method. In the end, the expanded uncertainty was presented. The results showed that the preparation of mixed standard solution from stock solution with precise pipettes had a relatively low uncertainty.

Computational interaction analysis of organophosphorus pesticides with different metabolic proteins in humans

Pesticides have the potential to leave harmful effects on humans, animals, other living organisms, and the environment. Several human metabolic proteins inhibited after exposure to organophosphorus pesticides absorbed through the skin, inhalation, eyes and oral mucosa, are most important targets for this interaction study. The crystal structure of five different proteins, PDBIDs： 3LII, 3NXU, 4GTU, 2XJ1 and 1YXA in Homo sapiens （H. sapiens）, interact with organophosphorus pesticides at the molecular level. The 3-D structures were found to be of good quality and validated through PROCHECK, ERRAT and ProSA servers. The results show that the binding energy is maximum -45.21 relative units of cytochrome P450 protein with phosmet pesticide. In terms of H-bonding, methyl parathion and parathion with acetylcholinesterase protein, parathion, methylparathion and phosmet with protein kinase C show the highest interaction. We conclude that these organophosphorus pesticides are more toxic and inhibit enzymatic activity by interrupting the metabolic pathways in H. sapiens.

Organophosphorus Pesticide Extraction and Cleanup from Soils and Measurement Using GC-NPD

The objectives of this study were to optimize instrumental parameters and conditions for analysis of selected organop- hosphorus pesticides (OPPs) by gas chromatography (GC) with nitrogen-phosphorus detection (NPD) (GC-NPD); to select an appropriate solvent system; to conduct a comparison of sonication and shaking extractions; and to select an appropriate procedure for extracting organophosphorus pesticides from soils. Procedure I consisted of n-hexane or petroleum ether together with acetone used as solvents, while Procedure contained several solvents including acetone, methanol, dichloromethane, and n-hexane or petroleum ether. Experimental results indicated that a mixture of petroleum ether/acetone (2:1, v/v) could be used in place of n-hexane/acetone (2:1, v/v) as it was a less expensive solvent system. In addition, shaking under a water bath at 20 °C was more effective than sonication. Also, Procedure I was more effective, safer, and more timesaving than Procedure Procedure I was applied to three soil types of different organic matter content, with recoveries of the OPPs from the yellow-brown soils, which had a higher organic matter content, being lower than those from the yellow and red soils.

Carboxylic Esterase and Its Associations With Long-term Effects of Organophosphorus Pesticides

To examine a） the effect of organophosphorus pesticide exposure on activity of carboxylic esterases, namely butyrylcholinesterase （BChE）, carboxylesterase （CarbE） and paraoxonase （PonE）; and b） the association of polymorphisms of BChE and PonE with individual genetic susceptibility to organophosphorus pesticide exposure. Methods A cross-sectional study was conducted in 75 workers exposed to organophosphorus pesticides and 100 non-exposed controls. The serum activity of these enzymes was measured. Variant forms of BCHE-K, PON-192, and PON-55 were detected. A symptom score was developed as a proxy measure of clinical outcomes. Results Activities of both BChE and CarbE were lower in exposed workers （27.3±21.65 runol.hl.mL^-l and 235.6±104.03 nmol-min^-l.mL^-l） than in non-exposed workers （78.313±30.354 nmol.h^-l.mL^-1 and 362.681_＋194.997 nmol.min^-1.mL^-1）. The activity of PonE was not associated with exposure status. The AChE activity in the exposed workers with BCHE-K genotype UU （61 cases）, genotype UK （12 cases） and genotype KK （2 cases） was 105.05, 84.42 and 79.00 mmol-h^-1.mL^-1, respectively and the accumulative symptom scores were 3.74, 9.17, and 12.50 accordingly. The AChE activity in the exposed workers with PON-192 genotype BB （37）, genotype AB （27） and genotype AA （11） was 116.8, 91.2, and 72,3 mmol-h^-1.mL^-1, respectively and the symptom scores were 2.00, 6.74, and 9.73 accordingly. The AChE activity in those with PON-55 genotype LL （70） and genotype LM （5） was 102.4 and 82.8 mmol-h^-1.mL^-1 and the symptom scores were 4.53 and 9.20. The symptom score was the highest in individuals with abnormal homozygote for each of the three gene loci. Condusions Long-term exposure to organophosphorus pesticides can inhibit BChE and CarbE activity, but exerts no inhibitory effect on PonE activity. Different genotypes of BCHE-K, PON-192, and PON-55 may be related to the severity of adverse health effects of organophosphorus pesticide exposure. Implications of potentially higher susceptibility of workers with mutant homozygotes should be evaluated to reduce health risks.

Application of Current Hapten in the Production of Broad Specificity Antibodies Against Organophosphorus Pesticides

Diethylphosphono acetic acid （DPA） was used as a current hapten to generate broad specificity polycolonal antibodies against a group of organophosphorus pesticides. Six New Zealand white rabbits were immunized with immunogens synthesized by the active ester method （AEM） or 1-ethyl-3-（3-dimethylaminopropyl）-carbodimide method （EDC）. The titers of antisera reached 25 600 by AEM and 6 400 by EDC, respectively. Polyclonal antibodies raised against DPA were screened and selected for the competitive indirect enzyme-linked immunosorbent assay （CI-ELISA）. A CI-ELISA for DPA was developed with a detection limit of 3.536 ng mL^-1and an I50 value of 0.182 μg mL^-1. The assay specificity was evaluated by obtaining competitive curves for several structurally related compounds as competitors. The antiserum showed high affinities to chlorpyrifos, diazinon, omethoate, parathion-ethyl and profenofos with I50 of 0.12, 0.15, 0.21, 0.88, 0.97 and 2.5 μg mL^-1, respectively. The results indicate that the assay could be a screening tool for quantitation and semiquantitation determination of the above former five organophosphorus pesticides.

The degradation of organophosphorus pesticides by means of sodium percarbonate

The factors and mechanisms of oxidative degradation of three organophosphorus pesticides （dichlorvos, methamidophos and phoxim） were studied with sodium percarbonate （SPC） as a solid oxidant. The result showed that SPC has highly activity in degrading these organophosphous pesticides. The most efficient degradation of pesticides occurred under basic conditions and the degradation rates increased with time extension and high temperature. The degradation of organophosphorus pesticides was expected to get even better results at lower initial concentration. Furthermore, we analyzed the intermediate products by NMP, spectrometry. On the basis of the analytical result, the oxidative degradation mechanism was proposed for each organophosphous pesticide. It is significant to understand the environment chemistry of organophosphorus pesticides in environmental system.

The Influencing Factors of Disposable Acetylcholinesterase Biosensor for in Situ Detection of Organophosphorus Pesticide

An amperometric biosensor based on acetylcholinesterase (ACHE) is assembled by simple adsorption of the AChE on 7,7,8,8-tetracyanoquinodimethane (TCNQ) modified screen-printed electrodes.This biosensor is used to detect the inhibitory effect of organophosphorus pesticides on AChE activity.The detection of organophosphorus pesticides is done with acetylthiocboline chloride (ATCh) as substrate.In order to obtain the optimized response to substrate,the influencing factors of the biosensor are investigated,including temperature,pH,incubation time,substrate concentration and AChE concentration. The measurements were performed after inhibition by immersing the enzyme electrode into the parathion methyl solution. Under the optimized conditions,that is,500 U/ml AChE concentration,pH 7.2,10 min incubation time,2 mmol/L substrate concentration and temperature of 37℃,from 5×10^(-8) mol/L to 5×10^(-5) mol/L is close to linear (regression equation: y(%)=124.055+15.7991gx,R^2=0.99644),which corresponds to 8.7%～56.1% AChE inhibition.With the optimized conditions, the lowest detectable amount of parathion methyl is 13×10^(-9).

Study of multiresidue analytical method for organonitrogen and organophosphorus pesticides in soil and water

A gas chromatographic method without derivatization was developed for the residue analysis of 10 organonitrogen and 9 organophosphorus pesticides in soil and water. The samples were blended or shaken with acetone for extraction. The extracts were cleaned up by coagulation, then, re-extracted with three 50 ml portions of dichloromethane. The final residue was detected by gas chromatography equipped with NPD. All of the 19 pesticides were completely separated at a constant temperature. The method described above was applicable to the simultaneous determination of 10 organonitrogen and 9 organophosphorus pesticides .in soil and water with the satisfactory recovery (from 82.42% to 103.57%), coefficient of variance (from 0.17% to 12.57%) and limit of detection (from 0.0006 ppm to 0.058 ppm).

Disposable Amperometric Acetylcholinesterase Biosensor for the Detection of Organophosphorus Pesticides

A rapid,simple,disposable and inexpensive acetylcholinesterase (ACHE) amperometric biosensor for the detection of organophosphorus pesticides was developed by simple adsorption of the enzyme on screen-printed electrodes.The biosensor consisted of an Ag/AgCl reference electrode and a graphite working electrode.The mixture of graphite and the 7,7,8,8-tetracyanoquinodimethane (TCNQ) was printed on electrodes.The detection of organophosphorus pesticides was done with acetylthiocholine chloride (ATCh) as substrate.The biosensor was used to detect the inhibitory effect of organophosphorus pesticides on AChE activity.The 1μl of enzyme solution containing 0.1 U AChE and 1% bovine serum albumin (BSA) were simply dropped on the working electrode surface.The biosensor operated at a potential of 300 mV vs. Ag/AgCl in a pH 7.2 0.1 mol/L phosphate buffer and 0.1 mol/L KCl.We obtained a calibration plot of the percentage inhibition versus the logarithm of parathion methyl concentration following an incubation time of 10 mix in parathion methyl solution. The lowest detectable amount of parathion methyl was 0.026 ppm.The amperometric biosensor based on acetylcholinesterase was disposable and low cost (about 1 yuan RMB).

Study on the toxic effect of the mixture of organophosphorus pesticide on perinatal rats

Objective To observe the toxic effect of the mixture of organophosphorus pesticide (MOP) on maternal rats and on the growth and development of their offspring. Methods Totally 40 Sprague-Dawley pregnant rats were randomly divided into three MOP dose groups and one control to which their offspring would be assigned. The experimental dosage of MOP and distilled water were administered orally starting on gestation day 15 and continued for 35 days. The physical development indices and the learning ability of F1 rats were measured during lactation. The pathological changes of uterus and liver of F0 rats were observed after weaning, while the weight ratio of uterus and some viscera to body of the F1 were examined. Results There were obvious changes of uterus and liver in the high-dose group of F0. The body-weight accretion of the F1 in high-dose group was obviously lower than that in control group (P<0.05). Some of the MOP F1 rats development indices delayed significantly (P<0.05), the learning ability decreased obviously, and the time of setting up memory prolonged (P<0.05). The ratio of the uterus weight to body-weight in the F1 MOP groups was significantly higher than that in control group (P<0.05). Conclusion The experiment doses of MOP are proved to have significant reproductive toxicity on perinatal rats.

Recent Advances in Degradation of Organophosphorus Pesticides

With the growing demand for environmental protection and physical health,food safety is now receiving more and more attention all over the world.However,pesticides are indispensable in agricultural production.Therefore,how to efficiently degrade pesticides and remove their residues in foods has always been a hot research topic in recent decades.This paper not only summarizes the types,degradation mechanism and artificial degradation of organophosphorus pesticides,but also highlights the latest advances in chemical degradation,photocatalytic degradation and biodegradation.

Toxicity of organophosphorus pesticide sumithion on larval stages of stinging catfish Heteropneustes fossilis

Sumithion is widely used to control brittle in paddy fields and tiger bug in fish larval rearing ponds. The objective of this study was to elucidate the toxic effects of sumithion on larval stages of stinging catfish Heteropneustes fossilis. Larvae were exposed to two concentrations(150 and 250 μg/L) of sumithion with one control in three replicates of each. Larvae samples were collected at 20- and 24-h intervals followed by observation under a digital microscope. Exposures of stinging catfish larvae to sumithion produced deformities including irregular head shape, lordosis, yolk sac edema, body arcuation, tissue ulceration, etc. The mortality rates of larvae were significantly increased in response to increase in sumithion concentrations. Furthermore, around 30% of the total adult stinging catfish reared in sumithiontreated aquaculture ponds were found to be deformed permanently. These findings highlight that exposure of stinging catfish to sumithion at the critical and sensitive stages in their life cycle may significantly reduce the number of returning adults. Therefore, the use of sumithion for crop protection needs to be considered carefully and alternatives to sumithion should to be developed for controlling aquatic insects in aqua-ponds during larval rearing.

The advantages of penehyclidine hydrochloride over atropine in acute organophosphorus pesticide poisoning:A meta-analysis

Background:Penehyclidine hydrochloride(PHC)has been used for many years as an anticholinergic drug for the treatment of acute organophosphorus pesticide poisoning(AOPP).The purpose of this meta-analysis was to explore whether PHC has advantages over atropine in the use of anticholinergic drugs in AOPP.Methods:We searched Scopus,Embase,Cochrane,PubMed,ProQuest,Ovid,Web of Science,China Science and Technology Journal Database(VIP),Duxiu,Chinese Biomedical literature(CBM),WanFang,and Chinese National Knowledge Infrastructure(CNKI),from inception to March 2022.After all qualified randomized controlled trials(RCTs)were included,we conducted quality evaluation,data extraction,and statistical analysis.Statistics using risk ratios(RR),weighted mean difference(WMD),and standard mean difference(SMD).Results:Our meta-analysis included 20,797 subjects from 240 studies across 242 different hospitals in China.Compared with the atropine group,the PHC group showed decreased mortality rate(RR=0.20,95%confidence intervals[CI]:0.16–0.25,P<0.001),hospitalization time(WMD=-3.89,95%CI:-4.37 to-3.41,P<0.001),overall incidence rate of complications(RR=0.35,95%CI:0.28–0.43,P<0.001),overall incidence of adverse reactions(RR=0.19,95%CI:0.17–0.22,P<0.001),total symptom disappearance time(SMD=-2.13,95%CI:-2.35 to-1.90,P<0.001),time for cholinesterase activity to return to normal value 50–60%(SMD=-1.87,95%CI:-2.03 to-1.70,P<0.001),coma time(WMD=-5.57,95%CI:-7.20 to-3.95,P<0.001),and mechanical ventilation time(WMD=-2.16,95%CI:-2.79 to-1.53,P<0.001).Conclusion:PHC has several advantages over atropine as an anticholinergic drug in AOPP.

Visible light assisted enzyme-photocatalytic cascade degradation of organophosphorus pesticides

The worldwide application of organophosphorus pesticides(OPs)has promoted agricultural development,but their gradual accumulation in soil and water can seriously affect the central nervous system of humans and other mammals.Organophosphorus hydrolase(OPH)is an effective enzyme that can catalyze the degradation of the residual OPs.However,the degradation products such as p-nitrophenol(p-NP)is still toxic.Thus,it is of great significance to develop a multi-functional support that can be simultaneously used for the immobilization of OPH and the further degradation of p-NP.Herein,a visible light assisted enzyme-photocatalytic integrated catalyst was constructed by immobilizing OPH on hollow structured Au-TiO_(2)(named OPH@H-Au-TiO_(2))for the degradation of OPs.The obtained OPH@H-Au-TiO_(2)can degrade methyl parathion to p-NP by OPH and then degrade p-NP to hydroquinone with low toxicity by using H-Au-TiO_(2)under visible light.OPH molecules were immobilized on HAu-TiO_(2)through adsorption method to prepare OPH@H-Au-TiO_(2).After 2.5 h of reaction,methyl parathion is completely degraded,and about 82.64%of the generated p-NP is further degraded into hydroquinone.After reused for 4 times,the OPH@H-Au-TiO_(2)retains more than 80%of the initial degradation activity.This research presents a new insight in designing and constructing multi-functional biocatalyst,which greatly expands the application scenarios and industrial value of enzyme catalysis.

Role of hemoperfusion combined with hemodialysis in acute severe organophosphorus pesticide poisoning:a meta-analysis

Background:Acute severe organophosphorus pesticide poisoning is a common severe emergency in developing countries.Our meta-analysis aimed to clarify the efficacy of hemoperfusion combined with hemodialysis on acute severe organophosphorus pesticide poisoning.Methods:Several databases were searched.After formulating relevant inclusion and exclusion criteria,qualified studies were included,and the data were extracted.The outcome indicators were the success rate of rescue,the time of hospitalization,the incidence of com-plications,the time for the cholinesterase level to return to normal,the coma time,and the atropine dosage.The results were analyzed using risk ratios,weighted mean difference,standard mean difference,and 95%confidence interval.The Cochrane Collaboration tool was used to assess the risk of bias in all the included studies.In terms of statistical methods,we used RevMan software(version 5.3;The Nordic Cochrane Centre,The Cochrane Collaboration,Copenhagen,Denmark)and Stata(versions 14 and 16;StataCorp,College Station,TX,USA)for the data analysis.Results:We included 92 randomized controlled trials with 6899 participants.Compared with the conventional emergency treatment group(CET group),the hemoperfusion group(HP+CET group),and the hemodialysis group(HD+CET group),the use of hemoperfusion combined with hemodialysis based on CET(HP+HD+CET group)significantly increased the success rate of rescue and reduced hos-pitalization time,the incidence of complications,time for cholinesterase level to return to normal,coma time,and atropine dosage.Conclusion:Hemoperfusion combined with hemodialysis is effective in the treatment of acute severe organophosphorus pesticide poi-soning.In the future,more scientifically designed,large-scale,high-quality,multicenter randomized controlled trials are needed to validate this study further.

Comparison of Two Procedures for Extraction and Clean-up of Organophosphorus and Pyrethroid Pesticides in Sediment

Two procedures were compared for extraction and clean-up of 20organophosphorus and 19 pyrethroid pesticides in sediment to identify the more effective procedurefor groups of pesticides or individual compounds. In Procedure I, methanol/water and n-hexane wereused for extraction, and 1:10 (v/v) dichloromethane in n-hexane and acetone were used as eluents foreluting the analyte through the cartridge, with one evaporating steps on a rotary evaporator andtwo eluting steps on the cartridge. n-hexane/acetone (2:1, v/v) was used for extraction and elutionin Procedure II with one evaporating step on a rotary evaporator and one eluting step on thecartridge. All extractions were performed under an ultrasonic bath and gas chromatography and massspectrometry were utilized for measurements. Procedure II was developed as a rapid, timesaving, lesscostly and safer substitute for Procedure I which was an old method. Procedure II was moreeffective for almost all the organophosphorus pesticides tested and 11of the 19 pyrethroidpesticides, while Procedure I was more appropriate for analysis of 5 pyrethroid pesticides. However,recoveries of most pyrethroid pesticides were fairly low. Thus, further studies should focus onadjustment and formulation of solvents for more efficient extraction and clean-up of pyrethroidpesticides from sediment samples.

Determination of organophosphorus pesticide residues in vegetables by an enzyme inhibition method using α-naphthyl acetate esterase extracted from wheat flour

The widespread use of organophosphorus pesticides(OPs) poses a great threat to human health and has made the detection of OP residues in food an important task,especially in view of the fact that easy and rapid detection methods are needed.Because OPs have inhibitory effects on the activity of α-naphthyl acetate esterase(ANAE) in plants,in this work we evaluated the possibility of detecting OPs in vegetables with ANAE extracted from commercial flour.The limits of detection(LODs) obtained for methamidophos,dichlorvos,phoxim,dimethoate,and malathion in lettuce samples with crude ANAE were 0.17,0.11,0.11,0.96,and 1.70 mg/kg,respectively.Based on the maximum residue limits(MRLs) for OPs in food stipulated by Chinese laws which are 0.05,0.20,0.05,1.00,and 8.00 mg/kg for methamidophos,dichlorvos,phoxim,dimethoate,and malathion,respectively,the esterase inhibition method with crude ANAE had sufficient sensitivity to detect the residues of dichlorvos,dimethoate,and malathion in lettuce,but it could not be used to guarantee the safety of the same samples if methamidophos or phoxim residue was present.The sensitivity of the method was improved by the use of esterase purified by ammonium sulfate salting-out.The LODs obtained for methamidophos and phoxim with purified esterase were lower than the MRLs for these OPs in food.This is a very promising method for the detection of OP residues in vegetables using crude or purified esterase because of its cheapness,sensitivity,and convenience.

Enzyme Inhibition Rate Method for Rapid Detection of Organophosphorus and Carbamate Pesticides in Cowpea

[Objectives ] The paper was to explore enzyme inhibition rate method for rapid detection of organophosphorus and carbamate pesticides in cowpea. [ Methods ] Acetylcholinesterase （ACHE） was added to cowpea extract, to determine the inhibition rate of extract against enzyme. The influences of different sampiing methods and sampling parts on detection results were compared. [ Results] The positive rate of standard sampling was 18.18% higher than that of non-stand- ard sampling, and the positive rate of samples collected from cowpea tail was 16.67% higher than that collected from other parts. [ Condmions] Enzyme inhibi- tion rate method is suitable for rapid detection of organophosphorus and carbamate pesticides in cowpea.