Low Concentrations Of Paraquat Research Articles

BRAP-2 promotes DNA damage induced germline apoptosis in C. elegans through the regulation of SKN-1 and AKT-1.

As part of the DNA damage response (DDR) network, the tumour suppressor Breast cancer susceptibility gene 1 (BRCA1) is activated to facilitate DNA repair, transcription and cell cycle control. BRC-1, the Caenorhabditis elegans ortholog of BRCA1, has conserved function in DNA double strand break repair, wherein a loss of brc-1 results in high levels of germline apoptosis. BRAP2/IMP was initially identified as a BRCA1 associated binding protein and previously we have shown that the C. elegans brap-2 deletion mutant experiences BRC-1 dependent larval arrest when exposed to low concentrations of paraquat. Since BRC-1 function in the germline is conserved, we wanted to determine the role of BRAP-2 in DNA damage induced germline apoptosis in C. elegans. We examined levels of germ cell death following DNA damage and found that brap-2(ok1492) mutants display reduced levels of germline apoptosis when compared to the wild type, and the loss of brap-2 significantly reduced germ cell death in brc-1 mutant animals. We also found increased mRNA levels of skn-1 following DNA damage in brap-2 mutants and that skn-1 RNAi knockdown in brap-2;brc-1 double mutants and a loss of pmk-1 mutation in brap-2 mutants increased apoptosis to wild type levels, indicating that brap-2 promotion of cell survival requires PMK-1 and SKN-1. Since mammalian BRAP2 has been shown to bind the AKT phosphatase PHLPP1/2, it suggests that BRAP2 could be involved in the Insulin/Insulin-like growth factor Signaling (IIS) pathway. We found that this interaction is conserved between the C. elegans homologs and that a loss of akt-1 in brap-2 mutants increased germline apoptosis. Thus in response to DNA damage, our findings suggest that BRAP-2 is required to attenuate the pro-cell survival signals of AKT-1 and PMK-1/SKN-1 to promote DNA damage induced germline apoptosis.

Root hydrotropism and thigmotropism in Arabidopsis thaliana are differentially controlled by redox status

ABSTRACTFactors that affect the direction of root growth in response to environmental signals influence crop productivity. We analyzed the root tropic responses of thioredoxin (trxs), thigmotropic (wav2-1), and hydrotropic (ahr1 and nhr1) Arabidopsis thaliana mutants treated with low concentrations of paraquat (PQ), which induces mild oxidative stress, and established a new method for evaluating root waviness (root bending effort, RBE). This method estimates root bending by measuring and summing local curvature over the whole length of the root, regardless of the asymmetry of the wavy pattern under thigmostimulation. In roots of the wav2-1 mutant, but not in those of the trxs and ahr1 mutants, RBE was significantly inhibited under mild oxidative stress. Thigmotropic stimulation of wav2-1 mutant roots, with or without PQ treatment, showed high levels of reactive oxygen species fluorescence, in contrast to roots of the ahr1 mutant. Furthermore, PQ inhibited root growth in all genotypes tested, except in the wav2-1 mutant. In a hydrotropism assay of the trxs and wav2-1 mutants, root growth behavior was similar to the wild type with and without PQ, while the root growth of ahr1 and nhr1 mutants was diminished with PQ. These results indicate that hydrotropic and thigmotropic mutants respond differently to exogenous PQ, depending on the tropic stimulus perceived. Therefore, the mechanisms underlying hydrotropism and thigmotropism may differ.

Paraquat inhibits cell viability via enhanced oxidative stress and apoptosis in human neural progenitor cells

Paraquat (PQ) is one of the most widely used herbicides in the world. Although available evidence indicates that people exposed to PQ have a higher risk of developing Parkinson’s disease, adverse effects of PQ on neural progenitor cells have not been investigated yet. In this study, we investigated the in vitro effect of PQ on immortalized human embryonic neural progenitor cells (hNPCs) by treating them with various concentrations of PQ (0, 0.1, 1, 10 and 100μM) for 24h. We show that PQ treatment reduces the cell viability and proliferation and induces reactive oxygen species (ROS) production in a dose-dependent manner. In addition, apoptosis induced by PQ was significantly increased at a concentration of as low as 1μM. To illustrate the underlying molecular mechanisms, we examined the caspase-3 activity, intracellular calcium level, the NF-κB activity, as well as expression of p21, p53 and metallothionein-III mRNA. PQ significantly increased caspase-3 activity at the concentration of 100μM. Similarly, PQ triggered intracellular Ca2+ releases and activation of NF-κB was observed after exposure of hNPCs at low concentrations of PQ (1μM). Meanwhile, p53 and p21 mRNA transcripts were significantly up-regulated at 10μM and 1μM of PQ, respectively. MT-III mRNA and protein expression was significantly up-regulated at 1μM of PQ and reached peak at 10μM. These results suggest that PQ could reduce viability of hNPCs by inducing oxidative stress and apoptosis.

An electrochemical magneto immunosensor (EMIS) for the determination of paraquat residues in potato samples

An electrochemical magneto immunosensor for the detection of low concentrations of paraquat (PQ) in food samples has been developed and its performance evaluated in a complex sample such as potato extracts. The immunosensor presented uses immunoreagents specifically developed for the recognition of paraquat, a magnetic graphite-epoxy composite (m-GEC) electrode and biofunctionalized magnetic micro-particles (PQ1-BSAMP) that allow reduction of the potential interferences caused by the matrix components. The amperometric signal is provided by an enzymatic probe prepared by covalently linking an enzyme to the specific antibodies (Ab198-cc-HRP). The use of hydroquinone, as mediator, allows recording of the signal at a low potential, which also contributes to reducing the background noise potentially caused by the sample matrix. The immunocomplexes formed on top of the modified MP are easily captured by the m-GEC, which acts simultaneously as transducer. PQ can be detected at concentrations as low as 0.18 ± 0.09 μg L(-1). Combined with an efficient extraction procedure, PQ residues can be directly detected and accurately quantified in potato extracts without additional clean-up or purification steps, with a limit of detection (90% of the maximum signal) of 2.18 ± 2.08 μg kg(-1), far below the maximum residue level (20 μg kg(-1)) established by the EC. The immunosensor presented here is suitable for on-site analysis. Combined with the use of magnetic racks, multiple samples can be run simultaneously in a reasonable time.

Effects of paraquat-induced oxidative stress on the neuronal plasma membrane Ca 2+-ATPase

Oxidative stress leads to the disruption of calcium homeostasis in brain neurons; however, the direct effects of oxidants on proteins that regulate intracellular calcium ([Ca 2+] i) are not known. The calmodulin (CaM)-stimulated plasma membrane Ca 2+-ATPase (PMCA) plays a critical role in regulating [Ca 2+] i. Our previous in vitro studies showed that PMCA present in brain synaptic membranes is readily inactivated by a variety of reactive oxygen species (ROS). The present studies were conducted to determine the vulnerability of PMCA to ROS generated in neurons as would probably occur in vivo. Primary cortical neurons were exposed to paraquat (PQ), a redox cycling agent that generates intracellular ROS. Low concentrations of PQ (5–10 μM) increased PMCA basal activity by two-fold but abolished its sensitivity to CaM. Higher concentrations (25–100 μM) inhibited both components of PMCA activity. Immunoblots showed the formation of high-molecular-weight PMCA aggregates. Additionally, PMCA showed evidence of proteolytic degradation. PMCA proteolysis was prevented by a calpain inhibitor, suggesting a role for calpain. Our findings suggest that PMCA is a sensitive target of oxidative stress in primary neurons. Inactivation of this Ca 2+ transporter under prolonged oxidative stress could alter neuronal Ca 2+ signaling.

Low Concentrations of Paraquat Induces Early Activation of Extracellular Signal-Regulated Kinase 1/2, Protein Kinase B, and c-Jun N-terminal Kinase 1/2 Pathways: Role of c-Jun N-Terminal Kinase in Paraquat-Induced Cell Death

Paraquat is a herbicide with a potential risk to induce parkinsonism due to its demonstrated neurotoxicity and its strong structural similarity to 1-methyl-4-phenylpyridinium (MPP(+)), a well-known neurotoxin which causes a clinical syndrome similar to Parkinson's disease (PD). However, at present very little is known about the signaling pathways activated by paraquat in any cell system. In this study, we have investigated the effect of paraquat on extracellular signal-regulated kinases 1 and 2 (ERK1/2), c-Jun N-terminal kinase (JNK), and protein kinase B (PKB) activation in E18 cells. Low concentrations of paraquat stimulated very early increases in ERK1/2, JNK1/2, and PKB phosphorylation. The phosphatidylinositol 3-kinase (PI-3K) inhibitors wortmannin and LY 294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one) inhibited early paraquat-induced increases in PKB phosphorylation. Furthermore, early paraquat-mediated increases in ERK1/2 activation were sensitive to the mitogen-activated protein kinase kinase 1 (MEK1) inhibitor PD 98059 (2'-amino-3'-methoxyflavone), whereas JNK1/2 responses were blocked by the JNK1/2 inhibitor SP 600125 (anthra[1-9-cd]pyrazol-6(2H)-one). Pretreatment with wortmannin, LY 294002, or PD 98059 had no effect on paraquat cell death in E18 cells. In contrast, SP 600125 significantly decreased paraquat-induced cell death in E18 cells. In conclusion, we have shown that low concentrations of paraquat stimulate robust very early increases in ERK1/2, JNK1/2, and PKB phosphorylation in E18 cells. Furthermore, the data presented clearly suggest that inhibition of the JNK1/2 pathway protects E18 cells from paraquat-induced cell death and support the fact that inhibition of early activation of JNK1/2 can constitute a potential strategy in PD treatment.

Induction of Phenotypes Resembling CuZn-Superoxide Dismutase Deletion in Wild-Type Yeast Cells: An in Vivo Assay for the Role of Superoxide in the Toxicity of Redox-Cycling Compounds

Yeast (Saccharomyces cerevisiae) lacking the enzyme CuZn-superoxide dismutase (sod1delta) display a large number of dioxygen sensitive phenotypes, such as amino acid auxotrophies, sensitivity to elevated temperatures, and sensitivity to 100% dioxygen, which are attributed to superoxide stress. Such cells are exquisitely sensitive to small amounts of the herbicide paraquat (methyl viologen), which is known to produce high fluxes of superoxide in vivo via a redox-cycling mechanism. We report that dioxygen sensitive phenotypes similar to those seen in sod1delta cells can be induced in wild-type cells by treatment with moderate concentrations of paraquat or diquat, another bipyridyl herbicide, providing strong evidence that the mechanism of toxicity for both of these compounds is attributable to superoxide stress. Certain redox-cycling quinone compounds (e.g., menadione and plumbagin) are also far more toxic toward sod1delta than to wild type. However, treatment of wild-type yeast with menadione or plumbagin did not induce sod1delta-like phenotypes, although toxicity was evident. Thus, their toxicity in wild type cells is predominantly, but not exclusively, due to mechanisms unrelated to superoxide production. Further evidence for a different basis of toxicity toward wild-type yeast in these two classes of redox-cycling compounds includes the observations that (i) growth in low oxygen alleviated the effects of paraquat and diquat but not those of menadione or plumbagin and (ii) activity of the superoxide sensitive enzyme aconitase is affected by very low concentrations of paraquat but only by higher, growth inhibitory concentrations of menadione. These results provide the basis for an easy qualitative assay of the contribution of redox-cycling to the toxicity of a test compound. Using this method, we analyzed the Parkinsonism-inducing compound 1-methyl-4-phenylpyridinium and found that redox cycling and superoxide toxicity are not the predominant factor in its toxic mechanism.

Paraquat-induced apoptotic cell death in cerebellar granule cells

We examined the toxicity of paraquat, a possible environmental risk factor for neurodegenerative disorders like Parkinson's disease (PD). Paraquat is structurally similar to the neurotoxin MPP + that can induce Parkinsonian-like features in rodents, non-human primates and human. Exposure of cerebellar granule cells to relatively low concentrations of paraquat (5 μM) produces apoptotic cell death with a reduction in mitochondrial cytochrome c content, proteolytic activation and caspase-3 activity increase and DNA fragmentation. Paraquat-induced apoptosis was significantly attenuated by co-treatment of cerebellar granule cells with the radical scavenger vitamin E, suggesting that paraquat-induced free radicals serve as important signal in initiation of cell death. As a decrease in mitochondrial cytochrome c content is also prevented by allopurinol, we suggest that xanthine oxidase plays an important role in the free radical production that precedes the apoptotic cascade and cell death after paraquat exposition.

Improved methods for the analysis of paraquat in biological fluids

Two methods are described for the detection of low concentrations of paraquat in biological fluids. A rapid spot test involves anin situ reaction on an SPE cartridge to form a vividly blue coloured radical ion. A quantitative method uses clean-up prior to oxidation to form the dipyridone derivative, which can be determined down to sub nanogram mL−1 levels using HPLC with fluorescence detection.

Comparative Effects of Paraquat on Antioxidant Components and Scavenging Enzymes in Kwangkyo and Hood Soybean

The effect of p araquaton hydroxyl radical production , glutathione and ascorbate content, and glutathione reductase and superoxide dismutase activities was assayed in the tolerant “Kwangkyo” and the susceptible “ Hood ” cultivars of soybean [Glycine max (L.) Merr.]. Seedlings of both cultivars at the fully expanded first trifoliate leaf stage were treated with paraquat at 0, 1, 10, 100, 500, and 1000 μᴍ. The in vivo generation of hydroxyl radicals was estimated by measuring methanesulfonic acid (MSA ) produced in soybean leaves treated with dimethylsulfoxide (DMSO ) and paraquat. The levels of paraquat-induced production of hydroxyl radicals were similar in Kwangkyo and Hood soybean. Ascorbate levels in the two cultivars were comparable and were reduced by paraquat treatment. Total glutathione levels were similar in both cultivars, but Kwangkyo had more glutathione in the reduced form than Hood. Treatment with low concentrations of paraquat enhanced oxidized glutathione levels in Hood. At high concentrations, paraquat decreased the levels of total and reduced glutathione in both cultivars. The endogenous activities of glutathione reductase and superoxide dismutase were similar in Kwangkyo and Hood soybean. Treatment with selected concentrations of paraquat enhanced the activity of these enzymes in trifoliates of the tolerant Kwangkyo. Overall, the results of this study do not suggest a strong involvement of components of the ascorbate-glutathione cycle in the observed tolerance of Kwangkyo soybean to the herbicide paraquat.

Paraquat poisoning: An analytical toxicologic study of three cases

Paraquat concentrations were measured in tissue, serum, urine and hemodialysate obtained from 3 patients who died 16.5 h, 22 days and 23 days after ingestion. In the patient who died 16.5 h post-ingestion, tissue paraquat levels were high. Kidney and liver had paraquat concentrations of 14 μg/g and 13.2 μg/g respectively, whereas lung tissue had a paraquat level of 3.8 μg/g. Low concentrations of paraquat were detectable in the tissues of the patients who died 22 and 23 days post-ingestion. Early in the poisoning, serum paraquat levels were high and large quantities of paraquat could be removed by both hemodialysis and forced diuresis. During an 8-h period, 713 mg of paraquat were removed by hemodialysis and 340 mg by forced diuresis. After the day of ingestion, little paraquat could be removed by hemodialysis or by forced diuresis; however, at all stages of the poisoning studied, hemodialysis was more effective than forced diuresis removing paraquat from the blood.

Chemical Curing of Range Grasses with Paraquat

Highlight Low concentrations of paraquat, when applied to range grasses at anthesis, effectively arrested the redistribution of nutrients in the herbage portions during curing. Treated herbage, high in estimated feed value and saved on the stump for later use by grazing animals may be another useful tool for the western livestock producer. The results warrant continued investigations with paraquat and other chemicals which may have similar characteristics when applied to grass herbage.