• Title/Summary/Keyword: organophosphate

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Cloning and Site-Directed Mutagenesis of Musca domestica Acetylcholinesterase for Enhancing Sensitivity to Organophosphorus and Carbamate Insecticides

  • Kim, Chung-Sei;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1760-1772
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    • 2006
  • Mature acetylcholinesterase (AChE) gene (gm, 1,836 bp) was cloned from the housefly and successfully expressed in the E. coli CodonPlus (DE3) RIL system (GM-E, 72 kDa) with a yield of 1,630 mU/g fresh cells. Using the gm, 10 kinds of mutants were constructed and expressed for enhancing sensitivity to insecticides. The sensitivity of these mutants to five kinds of organophosphate (OP) and three carbamate insecticides was investigated by measuring the apparent bimolecular inhibition constant ($k_i=k_2/K_d$). Surprisingly, the sensitivity of quadruple mutant IGFT was enhanced as much as 7-fold for acephate, 164-fold for demeton-S-methyl, 484-fold for dichlorvos, 523-fold for edifenphos, 30-fold for ethoprophos, 30-fold for benfuracarb, 404-fold for carbaryl, and 107-fold for furathiocarb, compared with that of GM-E, although the sensitivity of each single point mutant was slightly increased. These mutational studies indicated that (i) contradictory to Walsh et al. [39], the residue 327 is the important key residue for enhancing sensitivity as much as the residue 262, (ii) the residue 82 and additional residues of 234, 236, and 585 are also important, and (iii) sensitivity was cooperatively accelerated as the number of strategic mutations increased.

Regulation of Acetylcholine Esterase and Neurotransmitters in Oryzias latipes by Diazinon (다이아지는 처리에 의한 송사리의 아세틸콜린에스터라제 활성 및 신경전달물질 함량의 변화)

  • Kim, Jong-Sang;Koh, Sung-Cheol;Lee, Sung-Kyu;Chon, Tae-Soo
    • Environmental Analysis Health and Toxicology
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    • v.14 no.3
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    • pp.81-85
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    • 1999
  • Diazinon, an organophosphate pesticide, is relatively highly toxic to fish and causes vertebral malformation and behavioral change of fish at relatively low concentrations. To elucidate biochemical mechanism of the behavioral change of Oryzias latipes (killifish) caused by diazinon, the effect of the insecticide on acetylcholine esterase activities and the levels of some neurotransmitters were evaluated. Acetylcholine esterase activities in both head and body were significantly lowered at the concentration of 10 ppb of diazinon and acetylcholine contents in head tended to be upregulated with increasing concentration of diazinon. Exposure of killifish to 5000 ppb diazinon resulted in gradual decrease in acetylcholine content in body part with exposure time. Norepinephrine and serotonin concentrations in killifish head and body were highest at 1000 ppb of diazinon while neurotransmitter were relatively low in fish unexposed or exposed to lower dose of the pesticide, suggesting that increased norepinephrine and serotonin can partially account for diazinon-induced behavioral abnormality.

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Effects of Diethyl Phthalate on Acetylcholinesterase Activity in Olive Flounder (Paralichthys olivaceus) Following Short-term Exposure

  • Jee Jung-Hoon;Keum Yoo-Hwa;Kang Ju-Chan
    • Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.171-173
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    • 2004
  • Activity of acetylcholinesterase (AChE) is well known as a biomarker of exposure to organophosphate compounds in aquatic organisms. However, the effect of diethyl phthalate (DEP), a widely used plasticizer, on the chance of AChE activity is not yet known. Olive flounder (Paralichthys olivaceus) were exposed to DEP 300 and 1,000 mg DEP/kg b.w. through three times of intraperitoneal injection and effects were assessed in AChE activity of brain, muscle, heart and eyes of the exposed fish. AChE activity in various tissues of flounder was inhibited after exposure to DEP as a concentration-dependent manner, especially in brain, muscle and heart. Among tissues examined, heart is supposed to be a major part of body which is seriously damaged by DEP exposure. It indicates that DEP induces toxic effects in various organs (brain, muscle and heart), and changes of AChE activities. Such changed activities of AChE might be a useful biomarker to assess the impacts induced by phthalate esters including DEP.

Initial Blood Glucose Can Predict the Outcome of OP Poisoning (유기인계 중독환자에서 내원시 혈당과 예후와의 연관성)

  • Lee, Sung Do;Moon, Jeong Mi;Chun, Byeong Jo
    • Journal of The Korean Society of Clinical Toxicology
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    • v.13 no.2
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    • pp.55-61
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    • 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.

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Comparative effects of nicotine and diazinon on larval mortality and activity of cytochrome P-450 monooxygenases in Helicoverpa assulta and Spodoptera exigua (담배나방과 파밤나방의 유충사망률과 cytochrome P-450 monooxygenases의 활성에 미치는 니코틴과 다이아지논의 영향)

  • 이정호;부경생
    • Korean journal of applied entomology
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    • v.32 no.2
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    • pp.225-235
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    • 1993
  • This study was carried out to investigate effects of diazinon, an organophosphate, and nicotine, a plant~originated toxic chemical, on larval mortality and activity of cytochrome P-45D monooxygenases, a major detoxifwation enzyme system, in Helicoverpa assulta and Spodoptera exigua. Diazinon treatment gave a higher mortaliLy to H. assulta larvae than S. exigua larvae. In contrast to the case of diazinon, nicotine caused a higher mortahty to S. exigua than to H. assulla larvae. It was partly due to the fact that nicotine induced the actiVIty of midgut cytochrome P-450 monooxygenases (MFO) more than diazinon did in If assulta larvae. When If. aSSlllta larvae were reared on their host p\am. NicotwlIa tuoocum leaves, other componentS were mostly metabolized with the exception of dietary nicotine.

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Seizure-related Brain Injuries in Organophosphate Poisoning

  • Hur, Gyeung-Haeng;Lee,Yong-Soon;Han, Byung-Gon;Yeon, Gyu-Baek;Kim, Yun-Bae
    • Toxicological Research
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    • v.13 no.3
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    • pp.215-222
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    • 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.

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Determination of Cholinesterase Activities in Sera of Human and Different Species of Animals by Means of the Propionylthiocholine-Dithiobis Procedure (Propionylthiocholine-Dithiobis 방법에 의한 사람과 몇몇 동물 혈청내의 Cholinesterase 활성도 측정)

  • Kim, Soon-tae;Kim, Sung-hoon;Do, Jae-cheul;Lee, Young-ho;Mo, Ki-cheul;Huh, Rhin-sou
    • Korean Journal of Veterinary Research
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    • v.27 no.1
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    • pp.153-155
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    • 1987
  • Determination of cholinesterase activity is a routine practice in many laboratories to detect the influence of cholinesterase inhibiting drugs such as organophosphate and carbamate insecticides. Among many different methods to determine the cholinesterase activity, the present method was the most recent, simple and accurate one for routine test in clinics. The results obtained in sera of human and the different species of animals by means of the present method were as follows: $5.76{\pm}1.12U/ml$ in human, $3.37{\pm}0.83U/ml$ in german shepherd, $0.61{\pm}0.18U/ml$ in rat, $14.91{\pm}3.10U/ml$ in mouse, $1.55{\pm}0.51U/ml$ in chicken, $0.28{\pm}0.11U/ml$ in slaughtered cattle and $0.50{\pm}0.10U/ml$ in slaughtered pig.

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Microbial Degradation of Diazinon in Sudmerged Soil (담수토양내 미생물에 의한 Dazinon의 분해)

  • 김중호;이영하;최종우;이규승
    • Korean Journal of Microbiology
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    • v.27 no.2
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    • pp.139-146
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    • 1989
  • The mechanisms and metabolic products involved in the degradation of an organophosphate insecticide, diazinon, were studied in submerged paddy soil under the laboratory condition at $30^{\circ}C$. Diazinon abatement in non-sterilized soil was more rapid than indicating microbial participation in diazinon in soil. One-half of the original applications was lost in 2 days and less than 5% remained after 7 days. During the same period, dizinon applications increased tha microbial populations in accordance with the monooxygenase and esterase activities in soil. These results suggest that the microbiological factors develop in soil following diazinon application. The esterase and monooxygenase-catalyzing degradation products of diazinon were isolated and tentatively identified by mass spectrometryas 2-isopropyle-6-methyl-4-hydroxy pyrimidine, diazoxon, hydroxydiazinon, and sulfotep.

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Acute Immunotoxic Effects of Chlorpyrifos in CBA Male Mice (유기인계 Chlorpyrifos가 생쥐에 미치는 급성 면역 독성)

  • 김강석
    • Environmental Analysis Health and Toxicology
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    • v.13 no.1_2
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    • pp.33-41
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    • 1998
  • Chlorpyrifos, o,o diethyl-o-(3,5,6-trichloro-2-pyridyl) phosphorothioate, is a broad spectrum organophosphate insecticide. The use of chlorpyrifos has been increased more and more as pesticide. But the effects of chlorpyrifos on the immune alterations has not been yet observed. Therefore, we investigated the effects of chlorpyrifos on the immune alterations in CICA male mice. Chlorpyrifos was administered to mice by a single intraperitoneal injection for the purpose of observing acute effects. On the one hand to get the information on immunopathologic alterations we observed hematological values, counted total circulating leukocytes and assessed the ratio of lymphocytes and neutrophils from the peripheral blood, measured the ratio of organ/body weight and counted splenic cellularity in CBA male mice which treated chlorpyrifos intraperitoneally. But we could not find any significant immunopathologic alterations statistically by a single intraperitoneal injection. Also, the exposure of chlorpyrifos caused no significant change in the number of PFC/10$^6$ spleen cells at any three given doses. On the other hand a singte intraperitoneal injection of chlorpyrifos decreased the lymphocyte proliferation response slightly to ConA or LPS stimulation at a dose of 6 mg/kg b.w. Administrations of chlorpyrifos reduced mixed leukocyte response(MLR). MLR was decreased moderately at doses of 3mg/kg b.w. and 6mg/kg b.w. Therefore, all these findings suggest that chlorpyrifos may alter the immune functions acutely. especially by the changes of T lymphocyte activity.

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Heterostructured Nanophotocatalysts for Degradation of Organophosphate Pesticides from Aqueous Streams

  • Kaur, Paramjeet;Bansal, Priti;Sud, Dhiraj
    • Journal of the Korean Chemical Society
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    • v.57 no.3
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    • pp.382-388
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    • 2013
  • The present paper focuses on the synthesis, characterization and application of nanophotocatalyst for degradation of quinalphos and monocrotophos. Novel heterostructured ZnO/$TiO_2$ photocatalyst ($Z_9T$) was prepared and characterized with X-ray diffraction (XRD), SEM and UV-vis diffuses reflectance spectroscopy. The average crystalline size of synthesized $Z_9T$ was found to be 21.48 nm. The pesticides were degraded in the presence of nanophotocatalysts i.e., $TiO_2$, ZnO, $TiO_2$/ZnO mixed in various proportions and heterostructured nanophotocatalyst synthesized by Sol-Gel method. The batch experiments were performed by adding photocatalyst to 100 ml of pesticide solution and suspension was subjected to irradiation under UV light. In case of mixed catalyst, the maximum degradation of monocrotophos and quinalphos has been observed when ZnO and $TiO_2$ were in the ratio of 7:3 and 8:2 respectively. The degradation efficiency with synthesized heterostructured nanophotocatalyst ($Z_9T$) was found to be comparable with $TiO_2$.