Toxic effects of organophosphate pesticide monocrotophos in aquatic organisms: A review of challenges, regulations and future perspectives.

In recent times, usage of pesticide, herbicides and synthetic fertilizers in farming lands has made the environment worse. The pesticide residues and toxic byproducts from agricultural lands were found to contaminate the aquatic ecosystem. The misuse of synthetic pesticide not only affects the environment, but also affects the health status of aquatic organisms. The organophosphate pesticide pollutants are emerging contaminants, which threatens the terrestrial and aquatic ecosystem. Monocrotophos (MCP) is an organophosphate insecticide, utilized on crops including rice, maize, sugarcane, cotton, soybeans, groundnuts and vegetables. MCP is hydrophilic in nature and their solubilizing properties reduce the soil sorption which leads to groundwater contamination. The half-life period of MCP is 17-96 and the half-life period of technical grade MCP is 2500 days if held stable at 38 °C in a container. MCP causes mild to severe confusion, anxiety, hyper-salivation, convulsion and respiratory distress in mammals as well as aquatic animals. The MCP induced toxicity including survival rate, behavioural changes, reproductive toxicity and genotoxicity in different aquatic species have been discussed in this review. Furthermore, the ultimate aim of this review is to highlight the international regulations, future perspectives and challenges involved in using the MCP.

Biodegradation of monocrotophos by Brucella intermedia Msd2 isolated from cotton plant.

Widespread and inadequate use of Monocrotophos has led to several environmental issues. Biodegradation is an ecofriendly method used for detoxification of toxic monocrotophos. In the present study, Msd2 bacterial strain was isolated from the cotton plant growing in contaminated sites of Sahiwal, Pakistan. Msd2 is capable of utilizing the monocrotophos (MCP) organophosphate pesticide as its sole carbon source for growth. Msd2 was identified as Brucella intermedia on the basis of morphology, biochemical characterization and 16S rRNA sequencing. B. intermedia showed tolerance of MCP up to 100 ppm. The presence of opd candidate gene for pesticide degradation, gives credence to B. intermedia as an effective bacterium to degrade MCP. Screening of the B. intermedia strain Msd2 for plant growth promoting activities revealed its ability to produce ammonia, exopolysaccharides, catalase, amylase and ACC-deaminase, and phosphorus, zinc and potassium solubilization. The optimization of the growth parameters (temperatures, shaking rpm, and pH level) of the MCP-degrading isolate was carried out in minimal salt broth supplemented with MCP. The optimal pH, temperature, and rpm for Msd2 growth were observed as pH 6, 35 °C, and 120 rpm, respectively. Based on optimization results, batch degradation experiment was performed. Biodegradation of MCP by B. intermedia was monitored using HPLC and recorded 78% degradation of MCP at 100 ppm concentration within 7 days of incubation. Degradation of MCP by Msd2 followed the first order reaction kinetics. Plant growth promoting and multi-stress tolerance ability of Msd2 was confirmed by molecular analysis. It is concluded that Brucella intermedia strain Msd2 could be beneficial as potential biological agent for an effective bioremediation for polluted environments.

Organophosphorus Pesticides Promote Protein Cross-Linking.

Exposure to organophosphorus pesticides (OP) can have chronic adverse effects that are independent of inhibition of acetylcholinesterase, the classic target for acute OP toxicity. In pure proteins, the organophosphorus pesticide chlorpyrifos oxon induces a cross-link between lysine and glutamate (or aspartate) with loss of water. Tubulin is particularly sensitive to OP-induced cross-linking. Our goal was to explore OP-induced cross-linking in a complex protein sample, MAP-rich tubulin from Sus scrofa and to test 8 OP for their capacity to promote isopeptide cross-linking. We treated 100 µg of MAP-rich tubulin with 100 µM chlorpyrifos, chlorpyrifos oxon, methamidophos, paraoxon, diazinon, diazoxon, monocrotophos, or dichlorvos. Each sample was separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with Coomassie blue. Five gel slices (at about 30, 50, 150, and 300 kDa, and the top of the separating gel) were removed from the lanes for each of the eight OP samples and from untreated control lanes. These gel slices were subjected to in-gel trypsin digestion. MSMS fragmentation spectra of the tryptic peptides were examined for isopeptide cross-links. Sixteen spectra yielded convincing evidence for isopeptide cross-linked peptides. Ten were from the chlorpyrifos oxon reaction, 1 from dichlorvos, 1 from paraoxon, 1 from diazinon, and 3 from diazoxon. It was concluded that catalysis of protein cross-linking is a general property of organophosphorus pesticides and pesticide metabolites. Data are available via ProteomeXchange with identifier PXD034529.

Alterations in reproductive hormone levels among farm women and their children occupationally exposed to organophosphate pesticides.

Exposure to organophosphorus pesticides may lead to reproductive hormone dysfunction. Even among children of pubertal age, the exposure may disrupt growth, development, and maturation. The present study was conducted to assess the alterations in the reproductive hormone levels, among farm women (24-45 years, n = 129) and their children (9-12 years, n = 66 and 13-15 years, n = 63) and compare them with age and gender-matched control group [women (n = 134) and their children (9-12 years, n = 69 and 13-15 years, n = 65)] belonging to villages of Ranga Reddy District, Telangana, India. Blood pesticide residues and reproductive hormone (follicle-stimulating hormone-FSH, luteinizing hormone-LH, estradiol, and testosterone) levels were analyzed. The detected pesticide residues (ng/mL) were chlorpyrifos, diazinon, malathion, and monocrotophos among the farm women, while the farm children of 9-12 years age groups were detected with residues of chlorpyrifos, diazinon, malathion, monocrotophos, and phosalone. The farm children of 13-15 years age group were detected with residues of chlorpyrifos, diazinon, malathion, monocrotophos, and phosalone. However, no residues were detected among the samples of women and children of control groups. Significantly lower levels of FSH (in follicular phase) were observed among the farm women than the control group. Significant alterations in FSH and LH levels of farm women were observed with a significant correlation between the chlorpyrifos residue levels and estradiol hormone. While no such significant change in hormone levels was observed among the farm children of both age groups of both genders. Though the present study showed pesticide-induced alterations in hormone levels among the farm women, research is needed to elucidate the critical windows during which exposure may adversely affect the reproductive system in children at the pubertal stage and women at reproductive age and subsequently their progeny's health at a later stage of life.

N-acetylcysteine ameliorates monocrotophos exposure-induced mitochondrial dysfunctions in rat liver.

Background: Monocrotophos (MCP) is an organophosphate pesticide with well-known toxicity in mammals. Exposure of MCP is associated with altered molecular physiology at sub-cellular levels. This study investigated the efficacy of N-acetylcysteine (NAC) against MCP exposure mediated mitochondrial dysfunctions in hepatic tissue of rats.Methods: Male Wistar rats were given NAC (200 mg/kg b.wt), MCP (0.9 mg/kg b.wt) and NAC together with MCP, intragastrically for 28 consecutive days. Mitochondrial complexes activities were evaluated using biochemical analysis. mRNA expression of mitochondrial complexes subunits, PGC-1α and its downstream regulators were analyzed using polymerase chain reaction.Results: Exposure of MCP (0.9 mg/kg b.wt, intragastrically, 28 d) decreased mitochondrial complexes activities and gene expression of complexes subunits. The expression of PGC-1α, NRF-1, NRF-2, and Tfam was also reduced significantly. The administration of NAC (200 mg/kg b.wt, intragastrically, 28 d) significantly increased mitochondrial complexes activities and gene expression of complexes subunits. Additionally, NAC also maintained mitochondrial functions, and enhanced the gene expression of PGC-1α and its downstream regulators.Conclusion: The results of this study indicate that NAC prevents hepatic mitochondrial dysfunctions and maintains PGC-1α signaling. In conclusion, NAC might be speculated as a therapeutic agent for mitochondrial dysfunctions following toxic exposures.

Effects of in vitro exposure of perfluorooctanoic acid and monocrotophos on astroglia SVG p12 cells.

Glia cells provide supportive functions to the central nervous system and can be compromised by environmental contaminants. The primary objective of this study was to characterize the effects of in vitro exposure to perfluorooctanoic acid, a persistent environmental contaminant and/or monocrotophos (MCP), a neurotoxic organophosphate that is rapidly metabolized, to astroglia SVG p12 cells. The endpoints evaluated include cell viability, intracellular glutamate levels as a marker of astrocyte homeostasis function, differential gene expression for selected proteins, which include inflammatory markers (tachykinin), astrocytosis (nestin), S100B, and metabolism enzymes (CYP1A1). The results from cell viability revealed significant differences from the controls at some of the concentrations tested. Also, intracellular glutamate levels were elevated at the 10-µM concentration for perfluorooctanoic acid (PFOA) as well as the 10-µM PFOA/5-µM MCP concentration. Gene expression results at 80-µM PFOA concentration revealed a significant increase in the expression of S100B, tachykinin and CYP1A1. A combination of 10-µM PFOA/20-µM MCP caused a significant decrease in the expression of tachykinin. Gene expression for MCP exposures produced a decrease at the 20-µM MCP concentration. Immunofluorescence results indicated an increase in nestin protein expression for the 20-µM concentration of MCP, which contradicted the gene expression at the same concentration tested. The results indicate that toxicity to glia cells can compromise critical glia functions and could be implicated in neurodegenerative diseases.

Encapsulation of water soluble pesticides for extended delivery of pesticides without contaminating water bodies.

The main objective of this study is to develop polymeric encapsulated formulation for the water soluble broad-spectrum pesticides. Pesticides contaminate the environment in different ways but foremost hazards are linked with the contamination of water bodies. Water soluble pesticides are the major deleterious agents and go off into ground water and different water bodies through leaching or surface runoff from the applied places. Besides this some of the water soluble pesticides are broad-spectrum, but proper methods and techniques are not available for their effective and safe usage, all broad-spectrum pesticide are disappearing from the pesticide lists every year. Hence, the present study is based on development of encapsulated formulation for water soluble broad-spectrum pesticide i.e. Monocrotophos. In this study, water soluble pesticide was encapsulated in polyvinyl alcohol (PVA) polymer along with surfactants and cross linker. The developed microspheres were analyzed in HPLC for calculating loading capacity and encapsulation efficacy, these were calculated 0.75 and 90% respectively. The FT-IR data results confirmed that the monocrotophos successfully encapsulated in the PVA polymer with respective bands. The degradation studies show that in encapsulated formulation monocrotophos degradation was found only 10% after 94 hrs. Optical micrographs in aqeous solution indicate spherical shapes with size in the rage of 7-8 µm of encapsulated formulation. XRD data further crystalline nature of polymeric encapsulated formulation. The study may provide a new corridor to save the broad-spectrum water soluble pesticides which are on the verge to be banned.

DNA damage and biochemical responses in estuarine bivalve Donax incarnatus (Gmelin, 1791) exposed to sub-lethal concentrations of an organophosphate pesticide monocrotophos.

Monocrotophos (MCP) is a highly toxic and broad-spectrum pesticide extensively used for agricultural and household purposes. The present study was aimed to evaluate the genotoxicity and alterations in the biochemical and physiological conditions induced by monocrotophos in a non-target organism, an estuarine bivalve, Donax incarnatus. The bivalves were exposed to three sub-lethal concentrations (6.8, 13.7, and 27.45 ppm) of MCP for a period of 72 h. DNA damage was assessed using the comet assay. Oxidative stress was analyzed using catalase, glutathione peroxidase, and superoxide dismutase. Neurotoxicity was evaluated using the acetylcholinesterase assay (AChE) and the physiological condition was assessed using the condition index (CI). A significant concentration-dependent increase of DNA damage was observed as well as a decline in the activities of the antioxidant enzymes. However, a decrease in DNA damage was observed with advancing time. A significant decrease of AChE activity and CI was observed in the bivalves exposed to MCP. Positive correlations were also observed between DNA damage and the antioxidant enzymes whereas negative correlations were observed between AChE and the antioxidant enzymes indicating MCP toxicity mediated by oxidative stress.

Impact of pesticide monocrotophos on microbial populations and histology of intestine in the Indian earthworm Lampito mauritii (Kinberg).

Soil contamination has enlarged over the decades due to intensive use of pesticides and chemical fertilizers in agronomy. Earthworms are significant organisms in the soil community. Earthworms are the major role in soil fertility in most ecological system and the production of biogenic structures. Moreover, earthworm gut mucus enhances the beneficial soil microorganism potential biological activities. They are used as model organisms for assessing the ecological risks of chemicals. Enrichment of essential nutrients in soil through earthworm is a cost-effective and eco-friendly approach. In India, the organophosphorus pesticide monocrotophos is commonly used to control agricultural pests. Hence, it is important to study the effect of monocrotophos on the gut microbiota in Lampito mauritii. A 15-day exposure to a low (1/10th of the LC50 after 96 h i.e., 0.093 ppm kg-1) and high sublethal concentration (1/3rd of the LC50 after 96 h i.e., 0.311 ppm kg-1) of monocrotophos led to reduced proliferation of the gut microbiota in L. mauritii. However, exposure for 30 days led to a recuperation of the microbial populations to near control values. Among the eight bacterial and five fungal species that inhabit the gut of L. mauritii, only six bacterial and three fungal species were able to survive after exposure to monocrotophos. In addition to the study, histopathological changes were observed in the intestine of L.mauritii after application of lower sublethal concentration of monocrotophos. Severe pathological changes such as vacuolization, degenerated nuclei, damaged villi and congestion of the blood sinuses were noticed in the intestine on 1st and, 5th day of the experiment. But in 30th day the damages were slowly recovered due to degradation of monocrotophos by the presence of some pesticides degrading bacterial and fungal species and regenerative capability of chloragogen cells in the intestine. The results suggested that reduced microbial populations and pathological damages in intestine were observed during the application of monocrotophos. So, the monocrotophos have several harmful impacts on earthworms.

Deregulation of hepatic lipid metabolism associated with insulin resistance in rats subjected to chronic monocrotophos exposure.

In our previous study, we demonstrated the potential of monocrotophos (MCP), an organophosphorus insecticide (OPI), to induce glucose intolerance, insulin resistance (IR), and dyslipidemia with hyperinsulinemia in rats after chronic exposure. As hyperinsulinemia is likely to exert an impact on hepatic lipid metabolism, we carried out this study to establish the effect of chronic MCP exposure (0.9 and 1.8 mg/kg/day for 180 days) on hepatic lipid metabolism in rats. The state of IR induced by MCP in rats was associated with an increase in the liver lipid content (triglyceride and cholesterol) and expression levels of sterol regulatory element-binding proteins, PPARγ, acetyl-CoA carboxylase, and fatty acid synthase in the liver. Similarly, activities of key enzymes (acetyl-COA carboxylase, fatty acid synthase, lipin 1, malic enzyme, glucose-6-phosphate dehydrogenase, and glycerol-3-phosphate dehydrogenase), which regulate lipogenesis, were enhanced in livers of pesticide-treated rats. A strong correlation was observed between insulin levels, hepatic lipid content, and plasma lipid profile in treated rats. Our study suggests that long-term exposure to OPIs not only has a propensity to induce a state of hyperinsulinemic IR, but it is also associated with augmented hepatic lipogenesis, which may explain dyslipidemia induced by chronic exposure to MCP.

Acetylcholinesterase biosensors based on ionic liquid functionalized carbon nanotubes and horseradish peroxidase for monocrotophos determination.

Long-term and excessive use of monocrotophos (MPs) pesticide leads to an accumulation of MPs residues in agricultural products. Electrochemical biosensor technology was developed as a simple and efficient method for detecting MPs. However, commercial acetylcholinesterase (AChE) sensors are not applied in practical MPs detection due to poor stability and reliability. In this study, the advantages of functionalized carbon nanotubes (Cl/MWCNTs) and a bi-enzyme system (horseradish peroxidase (HRP)/AChE) were combined, a novel bi-enzyme electrode (Cl/MWCNTs/HRP/AChE/GCE) was constructed. Under optimal conditions, the bi-enzyme sensor had a wide detection range of 1.0 × 10-11 to 1.0 × 10-7 mol/L and low detection limit of 4.5 × 10-12 mol/L. The proposed AChE biosensor exhibited excellent stability and sensitivity for MPs determination and presented a promising tool for monitoring food safety.

Studies on the toxic effects of agrochemical pesticide (Monocrotophos) on physiological and reproductive behavior of indigenous and exotic earthworm species.

Earthworms are an ideal biological model in toxicity assays and environmental monitoring studies. In the present study, the reproductive toxicity and histopathological effects of Monocrotophos pesticide on an exotic epigeic Eudrilus eugeniae and an indigenous epigeic Perionyx barotensis earthworm were studied. Earthworm species were exposed to different concentrations of pesticide like 450 ppm, 500 ppm, and 650 ppm for 45 days and the mortality rate and reproductive activity was recorded every 15 days of exposure. There was an increase in mortality and abnormal sperm (asthenospermia, necrospermia, and oligospermia) and defective cocoons in earthworms with increasing concentrations of the pesticide. Histopathological changes like rupture of chloragogenous tissue, longitudinal muscle, fused and extra-villous growth and necrotic cell rupture in earthworm's body wall (epidermis, circular and longitudinal muscles) were observed. Fluorescent probes have detected cell death in pesticide-treated earthworms when compared to the control group after 45 days. The present findings show that Monocrotophos pesticide on exposure to epigeic earthworm species causes significant reproductive toxicity and histopathological abnormalities and these changes could be used as a tool in environmental risk assessment of pesticides.Abbreviations: DDT: Dichlorodiphenyltrichloroethane; MCP: Monocrotophos; EPA: Environment Protection Act; SL: Soluble Liquid; C: N (Carbon: Nitrogen); C: P (Carbon: Phosphorus); LC: Lethal Concentration; PBS: Phosphate Buffer Solution; WHO: World Health Organization; H&E: Hematoxylin and Eosin; SV: seminal vesicles; O: ovary; GP: genital papillae; Ch: chloragogenous tissue; EL: epithelial layer; CM: circular muscle; LM: longitudinal muscle; CD: cell debris.

Acetylcholinesterase with mesoporous silica: Covalent immobilization, physiochemical characterization, and its application in food for pesticide detection.

Toxic contamination of commonly consumed food products and water due to food chain vulnerability via agricultural products and commodities is a serious health hazard. This study reports on Santa Barbara Amorphous (SBA-15), a type of mesoporous silica nanoparticles, for efficient and stable acetylcholinesterase (AChE) adhesion toward detection of toxic pesticides. AChE was immobilized to the inert framework of mesoporous materials viz. SBA-15 with a proficient hydrolytic response toward acetylthiocholine. The immobilized system acts as a biosensor for the detection of pesticides, which are organophosphorus compounds in food. Both the SBA-15 and immobilized SBA-15 were characterized to give an insight on the physiochemical and morphological modification properties. The enzyme activity was accessed by Ellman's spectrophotometric bioassay for bare and enzyme-immobilized SBA-15 that resulted in promising enzymatic activity with the counterpart. Enzyme stability was also studied, which exhibited that immobilized AChE retained its catalytic activity up to 60 days and retained 80% of the hydrolytic activity even at 37°C. On the basis of the success of immobilized enzyme (covalent) being inhibited by acetylthiocholine, the sensor was administered for the inhibition by monocrotophos and dimethoate that are used widely as pesticides in agricultural. The inhibitory concentration (IC50 ) value was found to be 2.5 ppb for monocrotophos and 1.5 ppb for dimethoate inhibiting immobilized AChE. This was verified using cyclic voltammetry, an electrochemical analysis thus proving that the SBA-15@AChE complex could be used as a sensitive and highly stable sensor for detecting the concentration of hazardous pesticide compounds.

Monocrotophos detection with a bienzyme biosensor based on ionic-liquid-modified carbon nanotubes.

Acetylcholinesterase (AChE) biosensor technology is widely applied in the detection of organophosphate pesticides in agricultural production via the inhibition of AChE activity by organophosphates. However, the AChE electrode has some drawbacks, such as low stability and high overpotential. Combining the advantages of multiwalled carbon nanotubes (MWCNTs) and ionic liquids, we constructed a novel bienzyme electrode [Cl/iron porphyrin (FePP)-modified MWCNTs/AChE/glassy carbon electrode], which included AChE and mimetic oxidase FePP. In this electrode, FePP is covalently bound to the AChE carrier via ionic liquid for increased electrode sensitivity and stability. Under optimal conditions, this novel biosensor has a monocrotophos detection limit of 3.2 × 10-11 mol/L and good recovery of 89-104%. After 5 weeks of storage at 4 °C, the oxidation current was 97.8% of its original value. The biosensor has high stability and sensitivity for monocrotophos detection and is a promising device for monitoring food safety. Graphical abstract The complete synthesis process of Cl/FePP-MWCNTs/AChE/GCE.

Acetylcholinesterase biosensor based on functionalized surface of carbon nanotubes for monocrotophos detection.

Monocrotophos (Ops) has been widely used as pesticide in crop production but simultaneously could accumulate in the nature and seriously impact food safety and human health. It is necessary to develop a high sensitivity biosensor for accurate and fast detection of OPs. In this study, multi-walled carbon nanotubes (MWCNTs) were selected as acetylcholinesterase (AChE) carrier and their surface was modified by introducing different functional groups (-SH, -NH2, -Cl, -OH), hydrophobic alkyl groups (-CH3, -(CH2)2CH3, -(CH2)7CH3, -(CH2)15CH3) and ionic liquids (-IL1, -IL2). The interaction mechanism of MWCNTs functionalized surface and AChE has been revealed by studying characteristics of AChE immobilized on different carrier surface. Finally, compared with reported references and above other modifiers, we found that MWCNTs surface modified by -IL1 was the best carrier for AChE and the detection limit of IL1-MWCNTs/AChE/GCE was 3.3 × 10-11 M. At optimum reaction condition (pH 7.0, AChE loading 0.25 U, Inhibition time 14 min), storability test indicated reactivity of IL1-MWCNTs/AChE/GCE remained above 98.5% within two weeks. For real vegetable sample detection, the recoveries of IL1-MWCNTs/AChE/GCE were found to be between 90.0% and 104%. These results demonstrated novel biosensors could act as device of high sensitivity for accurate and fast detection of OPs.

Effects of monocrotophos pesticide on cholinergic and dopaminergic neurotransmitter systems during early development in the sea urchin Hemicentrotus pulcherrimus.

During early development in sea urchins, classical neurotransmitters, including acetylcholine (ACh), dopamine (DA), and serotonin (5-HT), play important roles in the regulation of morphogenesis and swimming behavior. However, the underlying mechanisms of how organophosphate pesticides cause developmental neurotoxicity by interfering with different neurotransmitter systems are unclear. In this study, we investigated the effects of 0.01, 0.10, and 1.00mg/L monocrotophos (MCP) pesticide on the activity of acetyltransferase (ChAT), acetylcholinesterase (AChE), monoamine oxidase, the concentration of DA, dopamine transporter, and the transcription activity of DA receptor D1 and tyrosine hydroxylase, during critical periods in cholinergic and dopaminergic nervous system development in sea urchin (Hemicentrotus pulcherrimus) embryos and larvae. At the blastula stages, MCP disrupted DA metabolism but not 5-HT metabolism, resulting in abnormal development. High ChAT and AChE activity were observed at the gastrulation-completed stage and the two-armed pluteus stage, respectively, MCP inhibited ChAT activity and AChE activity/distribution and resulted in developmental defects of the plutei. From the gastrula stage to the two-armed pluteus stage, we found ubiquitous disrupting effects of MCP on ACh, DA, and 5-HT metabolism, particularly at critical periods during the development of these neurotransmitter systems. Therefore, we propose that this disruption is one of the main mechanisms of MCP-related developmental neurotoxicity, which would contribute better understanding insight into the mechanism of MCP pesticide's toxic effects.

Curcumin revitalizes Amyloid beta (25-35)-induced and organophosphate pesticides pestered neurotoxicity in SH-SY5Y and IMR-32 cells via activation of APE1 and Nrf2.

Amyloid beta (Aß) peptide deposition is the primary cause of neurodegeneration in Alzheimer's disease (AD) pathogenesis. Several reports point towards the role of pesticides in the AD pathogenesis, especially organophosphate pesticides (OPPs). Monocrotophos (MCP) and Chlorpyrifos (CP) are the most widely used OPPs. In this study, the role of MCP and CP in augmenting the Aß-induced oxidative stress associated with the neurodegeneration in AD has been assessed in human neuroblastoma IMR-32 and SH-SY5Y cell lines. From the cell survival assay, it was observed that MCP and CP reduced cell survival both dose- and time-dependently. Nitro blue tetrazolium (NBT) based assay for determination of intracellular reactive oxygen species (ROS) demonstrated that Aß(25-35), MCP or CP produce significant oxidative stress alone or synergistically in IMR-32 and SH-SY5Y cells, while pretreatment of curcumin reduced ROS levels significantly in all treatment combinations. In this study, we also demonstrate that treatment of Aß(25-35) and MCP upregulated inducible nitric oxide synthase (iNOS/NOS2) whereas, no change was observed in neuronal nitric oxide synthase (nNOS/NOS1), but down-regulation of the nuclear factor erythroid 2-related factor 2 (Nrf2) level was observed. While curcumin pretreatment resulted in upregulation of iNOS and Nrf2 proteins. Also, the expression of key DNA repair enzymes APE1, DNA polymerase beta (Pol ß), and PARP1 were found to be downregulated upon treatment with MCP, Aß(25-35) and their combinations at 24 h and 48 h time points. In this study, pretreatment of curcumin to the SH-SY5Y cells enhanced the expression of DNA repair enzymes APE1, pol ß, and PARP1 enzymes to counter the oxidative DNA base damage via base excision repair (BER) pathway, and also activated the antioxidant element (ARE) via Nrf2 upregulation. Furthermore, the immunofluorescent confocal imaging studies in SH-SY5Y and IMR-32 cells treated with Aß(25-35) and MCP-mediated oxidative stress and their combinations at different time periods suggesting for cross-talk between the two proteins APE1 and Nrf2. The APE1's association with Nrf2 might be associated with the redox function of APE1 that might be directly regulating the ARE-mediated neuronal survival mechanisms.

Glucose-rich diet aggravates monocrotophos-induced dopaminergic neuronal dysfunction in Caenorhabditis elegans.

The present study aimed to obtain insights into the mechanism(s) by which glucose-rich diet aggravates monocrotophos (MCP)-induced dopaminergic neuronal dysfunction in Caenorhabditis elegans. In this study, we exposed three different strains of worms (wild-type N2, CB1112 (cat-2(e1112)II, tyrosine hydroxylase-deficient mutant, catecholamine absent) and the transgenic BZ555 (egls1-dat-1p::green fluorescent protein [GFP]) (in which bright GFP is tagged to the dopamine neuronal soma and processes) grown and maintained in normal nematode growth medium or 2% glucose enriched-nematode growth medium to MCP (0.75 mm) for 48 h. After the exposure, dopamine-mediated behaviors such as repulsion to nonanone, chemotaxis index and basal slowing response were determined in worms. Dopamine, 3,4-dihydroxy phenyl acetic acid and homovanillic acid content were quantified in N2 worms. The extent of neurodegeneration was visualized and quantified in dat-1::GFP worms. Basal slowing response study clearly indicated that cat-2 worms exposed to MCP and glucose were less affected compared to N2 of the same treatment. Learning and memory were affected by MCP and glucose. While MCP-treated worms showed lesser repulsion to nonanone compared to control worms, MCP-treated, glucose-fed worms showed a greater reduction in repulsion to nonanone. Further, MCP-treated, glucose-fed worms exhibited a marked reduction in dopamine content and an increase in 3,4-dihydroxy phenyl acetic acid and homovanillic acid levels compared to that in control. Dat-1::GFP showed a significant degeneration of dopaminergic neurons when exposed to glucose and MCP. Thus, our results clearly demonstrate that glucose-rich diet aggravates the dopaminergic neuronal dysfunction induced by MCP in C. elegans. Copyright © 2016 John Wiley & Sons, Ltd.

Phosphate-solubility and phosphatase activity in Gangetic alluvial soil as influenced by organophosphate insecticide residues.

An experiment was conducted under laboratory conditions to investigate the effect of four organophosphate insecticides, viz. monocrotophos, profenophos, quinalphos and triazophos at their field application rates (0.75, 1.0, 0.5 and 0.6 kg a.i.ha(-1), respectively), on the growth and activities of phosphate solubilizing microorganisms in relation to availability of insoluble phosphates in the Gangetic alluvial soil of West Bengal, India. The proliferation of phosphate solubilizing microorganisms was highly induced with profenophos (38.3%), while monocrotophos exerted maximum stimulation (20.8%) towards the solubility of insoluble phosphates in soil. The phosphatase activities of the soil (both acid phosphatase and alkaline phosphatase) were significantly increased due to the incorporation of the insecticides in general, and the augmentation was more pronounced with quinalphos (43.1%) followed by profenophos (27.6%) for acid phosphatase, and with monocrotophos (25.2%) followed by profenophos (16.1%) for alkaline phosphatase activity in soil. The total phosphorus was highly retained by triazophos (19.9%) followed by monocrotophos (16.5%), while incorporation of triazophos and quinalphos manifested greater availability of water soluble phosphorus in soil.

Adaptive response of rat pancreatic ß-cells to insulin resistance induced by monocrotophos: Biochemical evidence.

Our previous findings clearly suggested the role of duration of exposure to monocrotophos (MCP) in the development of insulin resistance. Rats exposed chronically to MCP developed insulin resistance with hyperinsulinemia without overt diabetes. In continuation of this vital observation, we sought to delineate the biochemical mechanisms that mediate heightened pancreatic ß-cell response in the wake of MCP-induced insulin resistance in rats. Adult rats were orally administered (0.9 and 1.8mg/kgb.w/d) MCP for 180days. Terminally, MCP-treated rats exhibited glucose intolerance, hyperinsulinemia, and potentiation of glucose-induced insulin secretion along with elevated levels of circulating IGF1, free fatty acids, corticosterone, and paraoxonase activity. Biochemical analysis of islet extracts revealed increased levels of insulin, malate, pyruvate and ATP with a concomitant increase in activities of cytosolic and mitochondrial enzymes that are known to facilitate insulin secretion and enhanced shuttle activities. Interestingly, islets from MCP-treated rats exhibited increased insulin secretory potential ex vivo compared to those isolated from control rats. Further, MCP-induced islet hypertrophy was associated with increased insulin-positive cells. Our study demonstrates the impact of the biological interaction between MCP and components of metabolic homeostasis on pancreatic beta cell function/s. We speculate that the heightened pancreatic beta cell function evidenced may be mediated by increased IGF1 and paraoxonase activity, which effectively counters insulin resistance induced by chronic exposure to MCP. Our findings emphasize the need for focused research to understand the confounding environmental risk factors which may modulate heightened beta cell functions in the case of organophosphorus insecticide-induced insulin resistance. Such an approach may help us to explain the sharp increase in the prevalence of type II diabetes worldwide.