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Applied and Environmental Microbiology Oct 2016Organophosphate pesticides used in agriculture can pose health risks to humans and wildlife. We hypothesized that dietary supplementation with Lactobacillus, a genus of...
UNLABELLED
Organophosphate pesticides used in agriculture can pose health risks to humans and wildlife. We hypothesized that dietary supplementation with Lactobacillus, a genus of commensal bacteria, would reduce absorption and toxicity of consumed organophosphate pesticides (parathion and chlorpyrifos [CP]). Several Lactobacillus species were screened for toleration of 100 ppm of CP or parathion in MRS broth based on 24-h growth curves. Certain Lactobacillus strains were unable to reach stationary-phase culture maxima and displayed an abnormal culture morphology in response to pesticide. Further characterization of commonly used, pesticide-tolerant and pesticide-susceptible, probiotic Lactobacillus rhamnosus strain GG (LGG) and L. rhamnosus strain GR-1 (LGR-1), respectively, revealed that both strains could significantly sequester organophosphate pesticides from solution after 24-h coincubations. This effect was independent of metabolic activity, as L. rhamnosus GG did not hydrolyze CP and no difference in organophosphate sequestration was observed between live and heat-killed strains. Furthermore, LGR-1 and LGG reduced the absorption of 100 μM parathion or CP in a Caco-2 Transwell model of the small intestine epithelium. To determine the effect of sequestration on acute toxicity, newly eclosed Drosophila melanogaster flies were exposed to food containing 10 μM CP with or without supplementation with live LGG. Supplementation with LGG simultaneously, but not with administration of CP 3 days prior (prophylactically), mitigated CP-induced mortality. In summary, the results suggest that L. rhamnosus may be useful for reducing toxic organophosphate pesticide exposure via passive binding. These findings could be transferable to clinical and livestock applications due to affordability and practical ability to supplement products with food-grade bacteria.
IMPORTANCE
The consequences of environmental pesticide pollution due to widespread usage in agriculture and soil leaching are becoming a major societal concern. Although the long-term effects of low-dose pesticide exposure for humans and wildlife remain largely unknown, logic suggests that these chemicals are not aligned with ecosystem health. This observation is most strongly supported by the agricultural losses associated with honeybee population declines, known as colony collapse disorder, in which pesticide usage is a likely trigger. Lactobacilli are bacteria used as beneficial microorganisms in fermented foods and have shown potentials to sequester and degrade environmental toxins. This study demonstrated that commonly used probiotic strains of lactobacilli could sequester, but not metabolize, organophosphate pesticides (parathion and chlorpyrifos). This Lactobacillus-mediated sequestration was associated with decreased intestinal absorption and insect toxicity in appropriate models. These findings hold promise for supplementing human, livestock, or apiary foods with probiotic microorganisms to reduce organophosphate pesticide exposure.
Topics: Animals; Caco-2 Cells; Chlorpyrifos; Drosophila melanogaster; Female; Humans; Lacticaseibacillus rhamnosus; Male; Organophosphates; Pesticides; Probiotics
PubMed: 27520820
DOI: 10.1128/AEM.01510-16 -
New Biotechnology Dec 2022In Escherichia coli, acyl carrier protein (ACP) is posttranslationally converted into its active holo-ACP form via covalent linkage of 4'-phosphopantetheine (4'-PP) to...
In Escherichia coli, acyl carrier protein (ACP) is posttranslationally converted into its active holo-ACP form via covalent linkage of 4'-phosphopantetheine (4'-PP) to residue serine-36. We found that the long flexible 4'-PP arm could react chemoselectively with the iodoacetyl group introduced on solid supports with high efficiency under mild conditions. Based on this finding, we developed site-selective immobilisation of proteins via the active holo-ACP fusion tag, independently of the physicochemical properties of the protein of interest. Furthermore, the molecular ratios of co-immobilised proteins can be manipulated because the tethering process is predominantly directed by the molar concentrations of diverse holo-ACP fusions during co-immobilisation. Conveniently tuning the molecular ratios of co-immobilised proteins allows their cooperation, leading to a highly productive multi-protein co-immobilisation system. Kinetic studies of enzymes demonstrated that α-amylase (Amy) and methyl parathion hydrolase (MPH) immobilised via active tag holo-ACP had higher catalytic efficiency (k/Km) in comparison with their corresponding counterparts immobilised via the sulfhydryl groups (-SH) of these proteins. The immobilised holo-ACP-Amy also presented higher thermostability compared with free Amy. The enhanced α-amylase thermostability upon immobilisation via holo-ACP renders it more suitable for industrial application.
Topics: Kinetics; Pantetheine; Acyl Carrier Protein; Escherichia coli; alpha-Amylases; Immobilized Proteins
PubMed: 36307012
DOI: 10.1016/j.nbt.2022.10.004 -
Biomolecules May 2024Olesoxime, a cholesterol derivative with an oxime group, possesses the ability to cross the blood-brain barrier, and has demonstrated excellent safety and tolerability...
Olesoxime, a cholesterol derivative with an oxime group, possesses the ability to cross the blood-brain barrier, and has demonstrated excellent safety and tolerability properties in clinical research. These characteristics indicate it may serve as a centrally active ligand of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), whose disruption of activity with organophosphate compounds (OP) leads to uncontrolled excitation and potentially life-threatening symptoms. To evaluate olesoxime as a binding ligand and reactivator of human AChE and BChE, we conducted kinetic studies with the active metabolite of insecticide parathion, paraoxon, and the warfare nerve agents sarin, cyclosarin, tabun, and VX. Our results showed that both enzymes possessed a binding affinity for olesoxime in the mid-micromolar range, higher than the antidotes in use (i.e., 2-PAM, HI-6, etc.). While olesoxime showed a weak ability to reactivate AChE, cyclosarin-inhibited BChE was reactivated with an overall reactivation rate constant comparable to that of standard oxime HI-6. Moreover, in combination with the oxime 2-PAM, the reactivation maximum increased by 10-30% for cyclosarin- and sarin-inhibited BChE. Molecular modeling revealed productive interactions between olesoxime and BChE, highlighting olesoxime as a potentially BChE-targeted therapy. Moreover, it might be added to OP poisoning treatment to increase the efficacy of BChE reactivation, and its cholesterol scaffold could provide a basis for the development of novel oxime antidotes.
Topics: Humans; Butyrylcholinesterase; Acetylcholinesterase; Ligands; Oximes; Cholinesterase Reactivators; Cholinesterase Inhibitors; Cholestenones; Kinetics; Sarin; GPI-Linked Proteins; Antidotes; Cholesterol; Organophosphorus Compounds
PubMed: 38785995
DOI: 10.3390/biom14050588 -
International Journal of Environmental... Aug 2022Pesticide-related mental health issues in Thailand, an upper-middle-income country, are not well known. This study aimed to investigate the association between the...
Pesticide-related mental health issues in Thailand, an upper-middle-income country, are not well known. This study aimed to investigate the association between the history of occupational exposure to pesticides and the mental health of Thai farmers. A cross-sectional study was carried out in the areas around Chiang Mai, a large city in Northern Thailand, between June 2020 and January 2021. A total of 6974 farmers from six districts were interviewed to determine whether they regularly experienced symptoms related to mental health by the Self-Reporting Questionnaire (SRQ-20) as well as their lifetime history of agricultural pesticide exposure from 31 active ingredients and five functional categories: insecticides, herbicides, fungicides, rodenticides, and molluscicides. The cut-off of 6 was used to evaluate probable mental disorder. Most of the farmers under investigation were men (53.8%), with a mean age of 55.2 (11.7) years, and were involved mainly in the planting of rice, fruit, and vegetables. About 86.7% reported having used pesticides on their crops at some point in their lives-mostly glyphosate, paraquat, 2,4-D, methomyl, and carbofuran. All functional groups, as well as pesticide classes like organochlorines, organophosphates, and carbamates, were significantly associated with a higher risk of probable mental disorder based on exposure duration, frequency, personal protective equipment usage, and hygienic behavior. In a model with multiple pesticides, there was an association between mental disorder and exposure to endosulfan (AOR = 2.27, 95%CI = 1.26-4.08) and methyl parathion (AOR = 2.26, 95%CI = 1.26-4.06). Having previously reported pesticide poisoning symptoms was related to mental disorder (AOR = 7.97, 95%CI = 5.16-12.31), the findings provided evidence of pesticide exposure posing a risk to farmers' mental health, particularly long-term and high-intensity exposure.
Topics: Agriculture; Cross-Sectional Studies; Farmers; Female; Health Status; Humans; Male; Middle Aged; Occupational Exposure; Pesticides; Thailand
PubMed: 35955007
DOI: 10.3390/ijerph19159654 -
Neurotoxicology Mar 2021Due to the increasing number of drugs and untested environmental compounds introduced into commercial use, there is recognition for a need to develop reliable and... (Comparative Study)
Comparative Study
Due to the increasing number of drugs and untested environmental compounds introduced into commercial use, there is recognition for a need to develop reliable and efficient screening methods to identify compounds that may adversely impact the nervous system. One process that has been implicated in neurodevelopment is neurite outgrowth; the disruption of which can result in adverse outcomes that persist later in life. Here, we developed a green fluorescent protein (GFP) labeled neurite outgrowth assay in a high-content, high-throughput format using induced pluripotent stem cell (iPSC) derived human spinal motor neurons and cortical glutamatergic neurons. The assay was optimized for use in a 1536-well plate format. Then, we used this assay to screen a set of 84 unique compounds that have previously been screened in other neurite outgrowth assays. This library consists of known developmental neurotoxicants, environmental compounds with unknown toxicity, and negative controls. Neurons were cultured for 40 h and then treated with compounds at 11 concentrations ranging from 1.56 nM to 92 μM for 24 and 48 h. Effects of compounds on neurite outgrowth were evaluated by quantifying total neurite length, number of segments, and maximum neurite length per cell. Among the 84 tested compounds, neurite outgrowth in cortical neurons and motor neurons were selectively inhibited by 36 and 31 compounds, respectively. Colchicine, rotenone, and methyl mercuric (II) chloride inhibited neurite outgrowth in both cortical and motor neurons. It is interesting to note that some compounds like parathion and bisphenol AF had inhibitory effects on neurite outgrowth specifically in the cortical neurons, while other compounds, such as 2,2',4,4'-tetrabromodiphenyl ether and caffeine, inhibited neurite outgrowth in motor neurons. The data gathered from these studies show that GFP-labeled iPSC-derived human neurons are a promising tool for identifying and prioritizing compounds with developmental neurotoxicity potential for further hazard characterization.
Topics: Cell Line; Genes, Reporter; Green Fluorescent Proteins; High-Throughput Screening Assays; Humans; Induced Pluripotent Stem Cells; Neural Stem Cells; Neuronal Outgrowth; Neurons; Risk Assessment; Toxicity Tests
PubMed: 33508353
DOI: 10.1016/j.neuro.2021.01.003 -
PloS One 2019We investigated a potential use of a 3D tetraculture brain microphysiological system (BMPS) for neurotoxic chemical agent screening. This platform consists of neuronal...
We investigated a potential use of a 3D tetraculture brain microphysiological system (BMPS) for neurotoxic chemical agent screening. This platform consists of neuronal tissue with extracellular matrix (ECM)-embedded neuroblastoma cells, microglia, and astrocytes, and vascular tissue with dynamic flow and membrane-free culture of the endothelial layer. We tested the broader applicability of this model, focusing on organophosphates (OPs) Malathion (MT), Parathion (PT), and Chlorpyrifos (CPF), and chemicals that interact with GABA and/or opioid receptor systems, including Muscimol (MUS), Dextromethorphan (DXM), and Ethanol (EtOH). We validated the BMPS platform by measuring the neurotoxic effects on barrier integrity, acetylcholinesterase (AChE) inhibition, viability, and residual OP concentration. The results show that OPs penetrated the model blood brain barrier (BBB) and inhibited AChE activity. DXM, MUS, and EtOH also penetrated the BBB and induced moderate toxicity. The results correlate well with available in vivo data. In addition, simulation results from an in silico physiologically-based pharmacokinetic/pharmacodynamic (PBPK/PD) model that we generated show good agreement with in vivo and in vitro data. In conclusion, this paper demonstrates the potential utility of a membrane-free tetraculture BMPS that can recapitulate brain complexity as a cost-effective alternative to animal models.
Topics: Acetylcholinesterase; Animals; Brain; Cell Line; Cell Survival; Imaging, Three-Dimensional; Mice; Microfluidics; Neurotoxins; Organophosphates; Receptors, GABA; Receptors, Opioid; Time Factors; Toxicity Tests
PubMed: 31703066
DOI: 10.1371/journal.pone.0224657 -
Journal of Toxicology 2014In recent years, the use of organophosphorus pesticides has been extensively increased and these compounds signify a major class of agricultural pesticides today. We...
Antigenotoxic Effect of Curcumin and Carvacrol against Parathion Induced DNA Damage in Cultured Human Peripheral Blood Lymphocytes and Its Relation to GSTM1 and GSTT1 Polymorphism.
In recent years, the use of organophosphorus pesticides has been extensively increased and these compounds signify a major class of agricultural pesticides today. We studied antigenotoxic potential of curcumin and carvacrol against the parathion induced DNA damage in cultured peripheral blood lymphocytes using sister chromatid exchanges as a biomarker of genotoxicity. Heparinised fresh blood from healthy individuals was treated with 2.5 μg/mL concentration of parathion in presence of curcumin and carvacrol in order to observe the antigenotoxic potential of both curcumin and carvacrol. Significant reduction (P < 0.05) was observed in the frequencies of SCEs in presence of 10 μg/mL and 15 μg/mL concentrations of curcumin as compared to parathion exposed sample. Similarly carvacrol had significant (P < 0.05) antigenotoxic effect at the concentrations of 2.5 μg/mL and 5.0 μg/mL against the parathion. We also studied the effect of GSTT1 and GSTM1 on genotoxicity of parathion and antigenotoxic potential of curcumin and carvacrol. We did not observe any significant effect (P > 0.05) of GSTT1 and GSTM1 polymorphism on genotoxicity of parathion and antigenotoxic potential of curcumin and carvacrol.
PubMed: 25328519
DOI: 10.1155/2014/404236 -
Aquatic Toxicology (Amsterdam,... Jan 2011Rainbow trout often serve as a surrogate species evaluating xenobiotic toxicity in cold-water species including other salmonids of the same genus, which are listed as...
Microsomal biotransformation of chlorpyrifos, parathion and fenthion in rainbow trout (Oncorhynchus mykiss) and coho salmon (Oncorhynchus kisutch): mechanistic insights into interspecific differences in toxicity.
Rainbow trout often serve as a surrogate species evaluating xenobiotic toxicity in cold-water species including other salmonids of the same genus, which are listed as threatened or endangered. Biotransformation tends to show species-specific patterns that influence susceptibility to xenobiotic toxicity, particularly organophosphate insecticides (OPs). To evaluate the contribution of biotransformation in the mechanism of toxicity of three organophosphate (phosphorothionate) insecticides, (chlorpyrifos, parathion and fenthion), microsomal bioactivation and detoxification pathways were measured in gills, liver and olfactory tissues in juvenile rainbow trout (Oncorhynchus mykiss) and compared to juvenile coho salmon (Oncorhynchus kisutch). Consistent with species differences in acute toxicity, significantly higher chlorpyrifos bioactivation was found in liver microsomes of rainbow trout (up to 2-fold) when compared with coho salmon. Although bioactivation to the oxon was observed, the catalytic efficiency towards chlorpyrifos dearylation (detoxification) was significantly higher in liver for both species (1.82 and 0.79 for trout and salmon, respectively) when compared to desulfuration (bioactivation). Bioactivation of parathion to paraoxon was significantly higher (up to 2.2-fold) than detoxification to p-nitrophenol in all tissues of both species with rates of conversion in rainbow trout, again significantly higher than coho salmon. Production of fenoxon and fenthion sulfoxides from fenthion was detected only in liver and gills of both species with activities in rainbow trout significantly higher than coho salmon. NADPH-dependent cleavage of fenthion was observed in all tissues, and was the only activity detected in olfactory tissues. These results indicate rainbow trout are more sensitive than coho salmon to the acute toxicity of OP pesticides because trout have higher catalytic rates of oxon formation. Thus, rainbow trout may serve as a conservative surrogate species for the evaluation of OP pesticides in coho salmon.
Topics: Analysis of Variance; Animals; Biotransformation; Chlorpyrifos; Chromatography, High Pressure Liquid; Fenthion; Gills; Insecticides; Microsomes, Liver; Olfactory Mucosa; Oncorhynchus kisutch; Oncorhynchus mykiss; Parathion; Species Specificity; Toxicity Tests
PubMed: 20947181
DOI: 10.1016/j.aquatox.2010.09.002 -
Heliyon Sep 2023Organophosphorus Pesticides () are among the extensively used pesticides throughout the world to boost agricultural production. However, persistent residues of these... (Review)
Review
Organophosphorus Pesticides () are among the extensively used pesticides throughout the world to boost agricultural production. However, persistent residues of these toxic pesticides in various vegetables, fruits, and drinking water poses detrimental health effects. Consequently, the rapid monitoring of these harmful chemicals through simple and cost-effective methods has become crucial. In such an instance, electrochemical methods offer simple, rapid, sensitive, reproducible, and affordable detection pathways. To overcome the limitations associated with electrochemical enzymatic sensors, non-enzymatic sensors have emerged as promising and simpler alternatives. The non-enzymatic sensors have demonstrated superior activity, reaching detection limit up to femto (10) molar concentration in recent years, leveraging higher selectivity obtained through the molecularly imprinted polymers, synergistic effects between carbonaceous nanomaterials and metals, metal oxide alloys, and other alternative approaches. Herein, this review paper provides an overview of the recent advancements in the development of non-enzymatic electrochemical sensors for the detection of commonly used OPPs, such as Chlorpyrifos (), Diazinon (), Malathion (), Methyl parathion () and Fenthion (). The design method of the electrodes, electrode functioning mechanism, and their analytical performance metrics, such as limit of detection, sensitivity, selectivity, and linearity range, were reviewed and compared. Furthermore, the existing challenges within this rapidly growing field were discussed along with their potential solutions which will facilitate the fabrication of advanced and sustainable non-enzymatic sensors in the future.
PubMed: 37662791
DOI: 10.1016/j.heliyon.2023.e19299 -
Frontiers in Public Health 2020Organophosphates (OP) are a major agrochemical. The application of OP pesticides is expected to increase multifold in the coming decades. The etiology of diabetic...
Organophosphates (OP) are a major agrochemical. The application of OP pesticides is expected to increase multifold in the coming decades. The etiology of diabetic diseases is attributed to multiple factors including OP pesticide exposure. The present study investigates pancreatic dysregulation with respect to exocrine enzymes and diabesity in groups of Pakistani and Cameroonian people exposed to a mixture of OP pesticides. Nine hundred and four OP exposed individuals were enrolled for this cross-sectional study after due consent and approval from an ethical review committee. Pesticides' residues were measured by GC-MS spectrometry. Cholinergic enzymes were measured by Elman's method. Serum glucose, insulin, serum amylase, lipase, and triglyceride were measured by spectrophotometry and ELISA; HOMA-IR was determined in OP exposed and non-exposed participants. Stata 15 and R 3.2.0 software were used for statistical analysis of the data. Malathion, chlorpyrifos, and parathion residues were evident in plasma samples. RBC-acetylcholinesterase was significantly depressed in OP exposed groups. In both population samples, investigated pancreatic functions were found to be statistically significantly more dysregulated than non-exposed. OP exposure indicated risk of diabetes and insulin, glycaemia, adiponectin, triglycerides, and TNF-α dysregulations. The study concludes that both OP exposed population groups exhibited a mixture of OP residues and pancreatic dysregulation, although the effect was more pronounced in the Cameroonian population. In addition, serum lipase has a positive correlation with OP exposure and diabetes and may be suggested as an alternate/additional diagnostic marker for diabesity under OP exposure. However, screening of other environmental co-factors with OP for pancreatic dysregulation is suggested.
Topics: Chlorpyrifos; Cross-Sectional Studies; Humans; Insecticides; Organophosphorus Compounds; Pesticides
PubMed: 33194944
DOI: 10.3389/fpubh.2020.534902