• Applied Biological Chemistry
• /
• 제41권4호
• /
• pp.213-217
• /
• 1998

An assay for cytochrome P450 monooxygenase activity by determination of the products of organophosphate oxidation via inhibition of acetylcholinesterase was described. Extracts from strains of Oryzaephilus surinamensis selected for resistance to chlorpyrifos-methyl (QVOS 102), fenitrothion (VOS F) and malathion (VOS 3), and a standard susceptible strain VOS 48, were incubated with an organophosphate in the presence of a NADPH-generating system and acetylcholinesterase. The degree of inhibition of the acetylchoinesterase activity was converted to manooxygenase activity using standard curves for the inhibition of acetylcholiesterase by chlorpyrifos-methyl-oxon, fenitrooxon and malaoxan. Activity was detectable in VOS 48 and was present at different increased levels with the different organophosphates in the three resistant strains, suggesting that different forms of P450 might be involved in organophosphate oxidation in these insects. The assays were carried out using crude insect homogenates and much smaller samples of insect material than the standard aldrin to dieldrin assay. It should be possible to use the method for determination of monooxygenase activity in single insert.

• 대한임상독성학회지
• /
• 제13권2호
• /
• pp.55-61
• /
• 2015

Purpose: Many studies have examined the mechanisms of impaired glucose homeostasis after organophosphate (OP) exposure, however no study has evaluated the clinical utility of blood glucose measurements in patients with OP poisoning. The current study was conducted to evaluate the initial glucose level at presentation and the glycemic variables during the first 3 days after admission as a predictor of mortality. Methods: This retrospective observational case series included 228 patients with a history of OP poisoning. Among other clinical data, information on the initial glucose level at presentation and mean glucose level, delta glucose level, and the presence of a hypoglycemic event during the first 3 days of admission, was collected. Results: Survivors had lower initial glucose levels at presentation and glucose variability during the first 3 days of admission compared to non-survivors. The frequency of hypoglycemic events was higher in non-survivors. In multivariate analysis, the initial glucose level (> 233 mg/dl) was an independent predictor of mortality, along with age. Conclusion: The initial glucose level at presentation can be helpful in prediction of mortality in cases of OP intoxication at bedside. The physician should pay attention to patients with a glucose level >233 mg/dl at presentation after ingestion of OP.

• Toxicological Research
• /
• 제13권3호
• /
• pp.215-222
• /
• 1997

The features of seizure-related brain injuries in rats poisoned i.p. with diisopropylfiuorophosphate were investigated. Pyridostigmine bromide (0.1 mg/kg) and atropine methylnitrate (20 mg/kg), which are centrally inactive, were pretreated i.m. 30 min and 10 min, respectively. before diisopropylfluorophosphate (10 mg /kg, $2LD_50$) poisoning to reduce the mortality and eliminate peripheral signs. Diisopropylfluorophosphate induced severe limbic seizures, and early necrotic and delayed apoptotic brain injuries. The necrotic brain injury, which was closely related to seizure intensity, was exerted as early as 1 hr predominently in hippocampus and piriform cortex. showing spongiform change (malacia) of neurophils in severe cases, in contrast to a typical apoptotic (TUNEL-positive)pattern after 12 hr in thalamus, and a mixed type in amygdala. Nitric oxide content in cerebrospinal fiuid significantly increased after 2 hr, reaching a maximal level at 6 hr. Pretreatment with $_L-N^G$-nitroarginine, an inhibitor of nitric oxide synthase, reduced nitric oxide content and attenuated only apoptotic brain injury in all four brain regions examined without affecting seizure intensity and necrotic injury. Taken together, early necrotic and delayed apoptotic brain injuries induced by diisopropylfiuorophosphate poisoning in rats may be related to seizure intensity and nitric oxide production, respectively.

• Journal of Applied Biological Chemistry
• /
• 제43권4호
• /
• pp.185-191
• /
• 2000

Pest insect control is dependent on about 200 insecticides that work by relatively few mechanisms. The targets they disrupt are mostly involved in the nervous system, respiratory chain, growth and development, or the gut. The major nerve targets are: acetylcholinesterase for the organophosphates and methylcarbamates; the nicotinic acetylcholine receptor for the neonicotinoids; the $\gamma$-aminobutyric acid receptor for several chlorinated hydrocarbons and fipronil; the voltage-gated sodium channel for DDT and pyrethroids. Selection of resistant strains often confers cross-resistance to some or all other insecticides working at the same site. The toxicological properties of different compounds acting on the same target are increasingly considered together, summating the risk even though the compounds are of quite diverse chemical types. Continuing attention is also being given to secondary targets not involved in the primary mechanism of toxicity but instead in side effects that must be considered in the overall safety evaluation. Research on insecticide targets is important in learning to keep up with resistance and changing concepts and policies on safety. These relationships are illustrated by recent studies in the Environmental Chemistry and Toxicology Laboratory of the University of California at Berkeley.

• 한국환경보건학회지
• /
• 제26권2호
• /
• pp.70-79
• /
• 2000

This study was done for the purpose of investigating the status of the use and perceptions of pesticides among the farmers who spray them in person. And it was also done to provide the basic statistics for the Agricultural Health Policy and study. Over the part of agricultural area in Kyungju and Ulsan City, Korea, from July to September 1999, 1032 questionnaires were distributed and 561 of them were collected and 447 were analysed. Major results of this study are as follows. The proportion of those who have special location for pesticides storage was only 5.3%, and that of those who lock hte storage location was only 13.6%. As to the treatment of bottles after use, most common response was 'burn in home'(47.7%). Most farmers purchase pesticides through 'pesticide store' or 'Agricultural Cooperatives'. The knowledge and Usage of pesticides depends in large part on experiences. The farmers who cultivate orchard spray pesticides more frequently than those cultivate paddy rice mainly. Most common pesticides in use are oganophosphates and carbamates. And also the pesticides that contains so-called 'Environmental Hormone' are used. The perception about the hazards of pesticides is rather superficial and insufficient. 'Herbicide(Gramoxon·Paraco)'(38%) is accepted as the most hazardous by farmers.

• Journal of Microbiology and Biotechnology
• /
• 제18권5호
• /
• pp.946-951
• /
• 2008

The opd gene, encoding organophosphorus hydrolase (OPH) from Flavobacterium sp. capable of degrading a wide range of organophosphate pesticides, was surface- and intracellular-expressed in Synechococcus PCC7942, a prime example of photoautotrophic cyanobacteria. OPH was displayed on the cyanobacterial cell surface using the truncated ice nucleation protein as an anchoring motif. A minor fraction of OPH was displayed onto the outermost surface of cyanobacterial cells, as verified by immunostaining visualized under confocal laser scanning microscopy and OPH activity analysis; however, a substantial fraction of OPH was buried in the cell wall, as demonstrated by proteinase K and lysozyme treatments. The cyanobacterial outer membrane acts as a substrate (paraoxon) diffusion barrier affecting whole-cell biodegradation efficiency. After freeze-thaw treatment, permeabilized whole cells with intracellular-expressed OPH exhibited 14-fold higher bioconversion efficiency ($V_{max}/K_m$) than that of cells with surface-expressed OPH. As cyanobacteria have simple growth requirements and are inexpensive to maintain, expression of OPH in cyanobacteria may lead to the development of a low-cost and low-maintenance biocatalyst that is useful for detoxification of organophosphate pesticides.

• Entomological Research
• /
• 제48권5호
• /
• pp.400-404
• /
• 2018

In this study, we investigated resistance to the organophosphates chlorpyrifos in Tunisian populations of Culex pipiens pipiens. Three field populations were collected from Northern and central Tunisia between 2003 and 2005 and used for the bioassays tests. Our results registered moderate and high levels of resistance to chlorpyrifos which ranged from 33.8 to 111. The chlorpyrifos resistant populations were highly resistant to propoxur indicated an insensitive acetylcholinesterase 1 (AChE 1). The highest frequency of AChE 1 resistant phenotypes (64%) was recorded in the most resistant population (sample # 1). Bioassays conducted in the presence of synergists showed that not esterases were involved as the resistance mechanism to chlorpyrifos. However, CYP450 was partly involved in the resistance of the most resistant sample (# 1). Starch electrophoresis showed that three esterases were present in studied samples: A2-B2, A4-B4 and B12. Results are discussed in relation to the selection pressure caused by insecticide treatments.

• 한국미생물·생명공학회지
• /
• 제51권4호
• /
• pp.374-389
• /
• 2023

Organophosphorus (OP) pesticides, particularly malathion, parathion, diazinon, and chlorpyrifos, are widely used in both agricultural and residential contexts. This refractory quality is shared by certain organ phosphorus insecticides, and it may have unintended consequences for certain non-target soil species. Bioremediation cleans organic and inorganic contaminants using microbes and plants. Organophosphate-hydrolyzing enzymes can transform pesticide residues into non-hazardous byproducts and are increasingly being considered viable solutions to the problem of decontamination. When coupled with system analysis, the multi-omics technique produces important data for functional validation and genetic manipulation, both of which may be used to boost the efficiency of bioremediation systems. RNA-guided nucleases and RNA-guided base editors include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR), which are used to alter genes and edit genomes. The review sheds light on key knowledge gaps and suggests approaches to pesticide cleanup using a variety of microbe-assisted methods. Researches, ecologists, and decision-makers can all benefit from having a better understanding of the usefulness and application of systems biology and gene editing in bioremediation evaluations.

• Journal of Microbiology and Biotechnology
• /
• 제21권1호
• /
• pp.71-80
• /
• 2011

Four diazinon-degrading bacteria were isolated from agricultural soil by using an enrichment technique. The biochemical analysis and molecular method including RFLP indicated that these isolates were identical, and one strain designated DI101 was selected for further study. Phylogenetic analysis based on 16S rDNA sequencing indicated that the strain DI101 clearly belongs to the Serratia marcescens group. The ability of the strain to utilize diazinon as a source of carbon and phosphorus was investigated under different culture conditions. The DI101 strain was able to completely degrade 50 mg/l diazinon in MSM within 11 days with a degradation rate of 0.226 $day^{-1}$. The inoculation of sterilized soil treated with 100 mg/kg of diazinon with $10^6$ CFU/g DI101 resulted in a faster degradation rate than was recorded in non-sterilized soil. The diazinon degradation rate by DI101 was efficient at temperatures from 25 to $30^{\circ}C$ and at pHs from 7.0 to 8.0. The degradation rate of diazinon was not affected by the absence of a phosphorus supplement, and addition of other carbon sources (glucose or succinate) resulted in the slowing down of the degradation rate. The maximum degradation rate ($V_{max}$) of diazinon was 0.292 $day^{-1}$ and its saturation constant ($K_s$) was 11 mg/l, as determined by a Michaelis-Menten curve. The strain was able to degrade diethylthiophosphate-containing organophosphates such as chlorpyrifos, coumaphos, parathion, and isazofos when provided as a source of carbon and phosphorus, but not ethoprophos, cadusafos, and fenamiphos. These results propose useful information for the potential application of the DI101 strain in bioremediation of pesticide-contaminated environments.

• 한국환경보건학회지
• /
• 제43권4호
• /
• pp.280-297
• /
• 2017

Objectives: We assessed pesticide exposure levels according to cultivation and crop type and investigated agricultural factors related to exposure. Methods: The participants, 341 male and 127 female farmers, were divided into three groups by cultivation crop type: chili greenhouse, cucumber greenhouse, and orchard. We collected questionnaires, socioeconomic characteristics and agricultural factors, and spot urine. Pesticide exposure was examined using four organophosphate and four pyrethroids urinary metabolites: dimethylphosphate, dimethylthiophosphate, diethylphosphate, diethylthiophosphate, Cis and Trans-3-(2-2dichlorovinyl)-2, 2-dimethylcyclopropane carboxylic acid, 3-phenoxybenzoic acid (3-PBA), Cis-3-(2-2dibrmovinyl)-2, and 2-dimethylcyclopropane carboxylic acid. Each metabolite was summed ${\Sigma}DAP$ and ${\Sigma}PY$ according to the chemical class. Results: Urinary metabolite detection rates and concentrations were similar between the greenhouse groups, but the orchard group was different. Similar 3-PBA detection rates were found in the three groups, but the geometric mean was very high in the orchard group compared to the two greenhouse groups. 3-PBA concentration in the orchard group was $4.11{\mu}g/g$ creatinine; the chili and cucumber greenhouse groups were 1.27 and $1.16{\mu}g/g$ creatinine, respectively. ${\Sigma}DAP$ was significantly associated with cultivation crop type and seasonal variation, but ${\Sigma}PY$ was not relevant. Conclusions: Our results suggest that cultivation and crop type may be correlated with different pesticide types and exposure levels. Furthermore, seasonal factors were related as potential factors influencing the level of organophosphate metabolites, but not for pyrethroid metabolites.