• Applied Biological Chemistry
• /
• v.42 no.4
• /
• pp.336-343
• /
• 1999

Fifty six mycelial cultured Basidiomycetes were screened for their inhibitory effects against prolyl endopeptidase(PEP), acetylcholine esterase(AChE) and thrombus coagulation. Out of them, methanolic extract of mycelium and/or ethylacetate(EtOAc) soluble fraction from culture broth of Peniophora quercina, Amanita aspera, Phellinus chrysoloma, Grifola frondosa, Wolfiporia extensa, Clavicorona pyxidata and Phanerochaete sordida inhibited more than 90% of PEP activity at 40 ppm. The extracts of Lenzites betulina, Phellinus chrysoloma, Wolfiporia extensa, Phanerochaete sorrlida, Hypocrea nigricans, Coriolus azureus, Flammulina velutipes, Phlebiopsis gigantea and Bondarzewia montana exhibited about 40% of inhibitory activity against AChE at 40 ppm. In thrombin times assay, the extracts of Amanita aspera, Oxyporus latemarginata, Peniophora quercina, Fomes fomenfarius, Trametes versicolor, and Phlebiopsis gigantea delayed coagulation of thrombus about two to three times over control at ca 550 ppm. In activated partial thromboplastin times assay, none of the tested Basidiomycetes showed significant effect.

• Korean Journal of Environmental Agriculture
• /
• v.7 no.2
• /
• pp.117-123
• /
• 1988

This study was initiated to understand the mechanism of selective toxicity of diazinon and carbofuran to killifish and loach. Conventional LC50 was calculated from fish test. IC50 with acetylcholinesterase activity was estimated using whole body and wet brain homogenate of the two fish species. Acetylcholinesterase activity of killifish was approximately twice as high as that of loach. The selective toxicity of diazinon to killifish and loach was partly (14 : 4) explained by the IC50 of diazoxon, a toxic metabolite of diazinon. IC50 of carbofuran also partly (14 : 3.4) contributed to the selectivity. These result suggested that the enzymatic method might be utilized as a screening tool for the chemicals affecting fish species of environmental concern with certain limitations which should be overcome in future studies.

• Environmental Analysis Health and Toxicology
• /
• v.17 no.3
• /
• pp.265-272
• /
• 2002

This study was carried out with the detection for multiresidue of the organophosphorus pesticides such as malathion, parathion. diazinon, and carbamate pesticide such as carbaryl, by enzyme-inhibition method. The acetylcholinesterase (AChE) and cholinesterase (ChE) activities in chicken brain determined by the Ellman's method were 166.6 and 5.8 $\mu$mol/min/g protein, and in chicken plasma were 23.1 and 8.3 $\mu$mol/min/g protein, respectively. The optimum pH of AChE and ChE was 8.2 and 7.8, respectively. The Km of AChE and ChE was 0.034 and 0.045 mM, respectively. I$\_$50/ for AChE and ChE by some organophosphorus was 55.82 and 99.42 mg/L of malathion, 31.16 and 29.13 mg/L of parathion, and 17.89 and 19.62 mg/L of diazinon, respectively. I$\_$50/ for AChE and ChE by carbaryl of carbamate was 0.10 and 0.05 mg/L, respectively. The 0.07 mg/L of drinking water advisory level for carbaryl could be detected with I$\_$50/ of AChE and ChE. Enzyme-Inhibition (EI) method with AChE and ChE was used the multiresidue method to detect the 1 mg/L of the carbamate pesticides.

• Journal of agriculture & life science
• /
• v.45 no.6
• /
• pp.213-226
• /
• 2011

This study was performed to investigate the effect of essential oils and ethanolic extracts of approximately 650 plant species on acetylcholine esterase (AChE) enzyme activity using Ellman's colorimetric method in 96-well microplates. The results showed that the ethanolic extracts from twig of Sophora subprostrata, twig of Phellodendron amurense, seed of Corylopsis coreana, and essential oil (EO) from Citrus paradisi, Cupressus sempervirens, Ocimum basilicum, Pinus sylvestris and Rosmarinus officinalis inhibited more than 80% of AChE activity. Among these, EO from Pinus sylvestris, C. sempervirens and C paradisi exhibited higher values of AChE inhibitory activity, which were 75, 84 and 99% at a concentration of 50 ug/ml, respectively. Finally, EO from C paradisi (grapefruit, GEO) showed the highest inhibitory activity towards AChE, which showed 91% of inhibition at a concentration of 20 ug/ml. We also examined the anti-dementia effects of GEO in mouse by passive avoidance test and Morris water maze test. The model mouse (male, ICR) of dementia (negative control) was induced by administration of scopolamine (1 mg/kg body weight). The latency time of sample group administrated with GEO (100 mg/kg, p.o.) increased significantly as compared with negative control on passive avoidance test. There were significant recovery from the scopolamine-induced deficits on learning and memory in water maze test through daily administrations with GEO (100 mg/kg, p.o.). From these results, we conclude that GEO treatment might enhance the cognitive function, suggesting that the EO of C. paradis may be a potential candidate for improvement of perceptive ability and dementia.

• The Korean Journal of Pesticide Science
• /
• v.7 no.1
• /
• pp.18-24
• /
• 2003

The activities of esterase and acetylcholinesterase(AChE) on the Diamond backmoth (DBM), Plutella xylostella (Lepidoptera: Yponomeutidae) and Beet armywarm (BAW), Spodoptera exigua (Lepidoptera: Noctuidae) and inhibitions of AChE with flupyrazofos were clarified from the results of a series of experiments. These findings are described in brief as follows. The AChE activities of DBM and BAW in heads were $1.5{\sim}11.1{\mu}$mol/g/min in $1st{\sim}4th$ instar larvae of DBM and $1.7{\sim}45.2{\mu}$mol/g/min in $1st{\sim}6th$ instar larvae of BAW, respectively. Those were $25{\sim}30$ times higher in above 4th instar larvae of BAW than that of the 1st instar larvae of DBM. The activities of aliesterase in heads were $1.7{\sim}4.7$ times higher in $2nd{\sim}4th$ instar larvae of DBM and $8{\sim}55$ times higher in $3rd{\sim}6th$ instar larvae of BAW than 1st instar larvae of DBM. In abdomens, those were $3{\sim}17$ times higher in $2nd{\sim}4th$ instar larvae of DBM and $12{\sim}30$ times higher in $3rd{\sim}6th$ instar larvae of BAW than 1st instar larvae of DBM. Median AChE inhibition concentration $(I_{50})$ of flupyrazofos to the 2nd instar larvae of DBM and BAW were 92 nM and $15{\mu}M$, respectively, and those to the 4th instar larvae of DBM and BAW were $1.8{\mu}M$ and 3.1 mM, respectively. Insensitivity ratio of flupyrazofos in the 2nd instar BAW larvae showed ca. 162 times higher than that in the 2nd instar larvae of DBM, and that of the 4th instar BAW larvae showed ca. 1,720 times higher insensitivity to flupyrazofos than that of the 4th instar DBM larvae. AChE activities in the 2nd instar larvae of DBM and BAW at 32 h after applicaton of flupyrazofos decreased from 67.6% to 32.4% of the activity of the untreated control. That of the 4th instar larvae of DBM increased for 0.5 h after application flupyrazofos up to 75% of the untreated control, and after that it decreased to 34.5% of the untreated control at 32 h. In contrast, in the 4th instar larvae of BAW AChE activities increased for 8 h gradually up to 102 % of the activity of the untreated control, and then the activity decreased to 97% of the untreated control at 16 h after treatment.

• The Korean Journal of Pesticide Science
• /
• v.7 no.1
• /
• pp.45-50
• /
• 2003

This study was conducted to understand the role of oxidative enzyme cytochrome $P_{450}$ in the bioactivation of benfuracarb and to know metabolites of benfuracarb by cytochrome $P_{450}$. The bimolecular imhibition rate constant $(k_i)$ of benfuracarb on acetylcholinesterase (AChE) was as low as $1.1{\times}10^3\;M^{-1}\;min^{-1}$, suggesting that benfuracarb should be activated for its toxic action. The potency of benfuracarb on AChE in the oxidase system (cytochrome $P_{450}$ + NADPH) in vitro was 10-fold higher than that of control (cytochrome $P_{450}$). Such a similar result was also found in the oxidase + PBO system. In vivo the $I_{50}$ of benfuracarb was 22.7mg $kg^{-1}$, but pie-treatment of piperonyl butoxide (PBO) reduced the $I_{50}$ by >100mg $kg^{-1}$. This result suggests that cytochrome $P_{450}$ was involved in the activation of benfuracarb. Using microsomal oxidase system, metabolites of benfuracarb were elucidated. Fifty-eight percent of benfuracarb was converted to carbofuran, a major toxic metabolite, in the oxidase system, while only less than two percent of benfuracarb was converted to carbofuran in the oxidase + PBO system. These results also suggest that cytochrome $P_{450}$ was involved in the activation of benfuracarb. Overall results indicate that cytochrome $P_{450}$ could be involved in the bioactivation of benfuracarb to carbofuran.

• The Korean Journal of Pesticide Science
• /
• v.9 no.1
• /
• pp.97-101
• /
• 2005

Flupyrazofos (O,O-diethyl O-1-phenyl-3-trifluoromethylpyrazo-5-yl phosphorothioate) is an organophosphorus insecticide with a pyrazole moiety which is newly developed and commercialized by SUNGBO chemical company and Korean Research Institute of Chemical Technology for effectively control against diamond back moth. This study was conducted to determine the composition and quantity of impurities in technical 1 (94.5%), technical 2 (97.6%) and purified (99.2%) flupyrazofos using GLC/MSD. Bimolecular inhibition rate constant($k_i$) with acethylcholinesterase (in vitro) and $I_{50}$ with mouse brain acetylcholinesterase (in vivo) were measured for comparing inhibitory patterns of two technicals and purified flupyrazofos. Impurities of flupyrazofos were identified as O,O,O-triethylthio-phosphoric acid (TEA), 1-phenyl-3-trifluoromethyl-5-ethoxy pyrazole(PTMEP), O,O-diethyl O-1-phenyl-3-trifluoromethylpyrazo-5-yl phosphoric acid ester(flupyrazofos oxen), O,S-diethyl O-1-phenyl-3-trifluoromethylpyrazo-5-yl phosphorothionate (S-ethyl flupyrazofos). In in vitro, technical 1 showed the fastest inhibition on AChE activity among them. And technical 1 and 2 showed 40% higher in vivo inhibition against mouse brian AChE than purified flupyrazofos did. These results could be caused by the impurities such as flupyrazofos oxen and S-methyl flupyrazofos contained in technical grades of flupyrazofos.

• The Korean Journal of Pesticide Science
• /
• v.7 no.4
• /
• pp.288-295
• /
• 2003

To determine the mechanism of the resistance to organophosphorus insecticide, chlorpyrifos, in diamondback-moth (Plutella xylostella L.), activities of esterases, glutathione-S-transferase (GST) and AChE insensitivity which were known for causing factor of resistance were measured. Also, the relationship between AChE insensitivity and the resistant ratio was investigated to inquiry the cross-resistance. The resistant ratio of chlorpyrifos-resistant strain (CRS) of diamondback-moth at the 6th generation was developed 160 fold compared to susceptible strain (SS) one. Activity of GST that are extracted from CRS was 1.7-fold higher than that from SS. However, activity of total esterases from CRS was similar to that from SS. In AChE insensitivity test, CRS was 11.8-fold less sensitive than that from SS. CRS was ranged from 17.6 to 33.6-fold less sensitive than SS to other insecticides having same target site with chlorpyrifos such as dichlorvos, dimethylvinphos and carbofuran. Insensitivity of AChE to phenthoate-oxon, however, was 1.7-fold. Resistance of CRS was 82-fold, 47-fold and 42-fold higher than SS to dichlorvos, dimethylvinphos and carbofuran, respectively, but 2.3-fold to phenthoate and then we could identify that the resistance development of insecticide might have a lot of difference among the chemicals with the same target site. The relationship between the AChE insensitivity and the resistant ratio was significantly correlated$(r=0.9951^{**},\;p^{(0.01)}$. This result indicates that AChE insensitivity was associated with insecticide resistance. Overall, these results suggest that insensitivity of AChE was an important factors to chlorpyrifos resistance in diamondback-moth, and the slightly increased activity of GST may also have contributed to that.

• Korean journal of applied entomology
• /
• v.30 no.1
• /
• pp.18-28
• /
• 1991

The optimal pH of the extraction buffer was 7.5 considiering AChE stability and its buffer capacity when AChE was isolated and extracted from the housefly(Musca domesitca L.)and three other insect species with 0.01 M sodium phosphate buffer. Also, the optimal pH of the reaction buffer was 7.5 considering enzyme activity and its buffer capacity when AChE activity was measured with the substrate in 0.1 M sodium phosphate buffer. The Potter Elvehjem type homogenizer with Teflon pestle was used to homgenize the tissues. When preparing a AChE suspension by centrifuging the homogenate, 700 g supernatant of adult head for the housefly, 700 g supernatnat of 5th instar nymphal whole body for the brown planthopper, lipid-eliminated 10,000 g supernatant of 5th instar larval whole body for the diamondback moth, and 700 g supernatant of 4th instar larval head for the tobacco cutworm were considered satisfactory as enzyme sources in view of mass preparation, extraction efficiency and stability of enzyme activity during evaluation. When AChE suspensions of 4 insect species were stored at $-18^{\circ}C$, more than 90% of activity was maintained up to 3 weeks. Km values of AChEs of the housefly, the brown planthopper, and the diamondback moth were 0.042, 0.037 and 0.043 mM, respectively and AChE-specific substrate inhibition was observed at high concentration. Km value of the tobacco cutworm ChE was 1.15 mM and BuChE characteristics was observed, though further study is needed. The optimal substrate concentration for the AChE inhibition tests was 0.5 mM for the housfly, the brown planthopper, and the diamondback moth and 12 mM for the tobacco cutworm.

• Journal of Acupuncture Research
• /
• v.25 no.2
• /
• pp.1-9
• /
• 2008

목적 : Acetylcholine은 콜린과 아세트산의 에스테르로 인체에서 중요한 신경전달물질로 Acetylcholine-sterase(AChE)라는 효소에 의해 분해된다. Alzheimer's disease, ataxia, myasthenia gravis, Parkinson' disease 등의 질환에 AChE 억제제가 사용되어 왔으며 최근 한약재의 AChE 억제 효능에 관한 연구들도 진행되고 있다. 봉독은 관절염, 통풍 등의 질환에 응용되어 왔으며 진통효과 및 항염증작용에 대한 임상적, 실험적 연구가 많이 보고되어 왔으나 AChE 억제효과에 대한 연구는 아직까지 보고된 바 없다, 본 연구에서는 봉독약침액과 봉독의 과민반응 유발항원 중 하나인 Phospholipase A2 억제효능이 있는 것으로 알려진 상백피를 혼합한 상백피봉독약침액의 AChE 억제효과를 알아보았다. 방법 : PC12 세포주에서 추출한 AChE와 0.1, 0.01 and $0.001mg/m{\ell}$ 농도의 봉독약침액 및 상백피봉독약침액을 60분간 반응시켰다. 효소면역측정법(ELISA)을 이용하여 흡광도를 10분, 30분, 60분 경과시 각각 측정한 후 효소활성저해도(%)를 계산하였다. 효소활성저해도(%) = [(Cc - Ce)/Cc] ${\times}$ 100 Cc : 대조군 흡광도, Ce : 실험군 흡광도 결과 : 1. 봉독약침액은 0.1, 0.01, $0.001m{\ell}/mg$의 농도에서 30분 경과 후부터 유의성 있는 억제효과를 나타내었다. 2. 상백피봉독약침액은 $0.1m{\ell}/mg$ 농도에서 10분 경과 후부터 유의성 있는 억제효과를 나타내었고, $0.01m{\ell}/mg$ 농도에서 30분경과 시 유의성 있는 억제효과를 나타내었다. 3. 봉독약침액과 상백피봉독약침액의 AChE 억제효과 비교에서 봉독약침액의 억제효과가 상백피봉독약침액 보다 뛰어났다. 요약 : 봉독약침액과 상백피봉독약침액의 AChE 억제효과를 확인하여 두 군 모두 유의성 있는 결과를 얻을 수 있었다. 앞으로 알츠하이머병이나 치매와 같은 신경퇴행성 질환에 대한 봉독의 임상적 활용 및 보다 넓은 범위에 대한 연구가 필요할 것이라 사료된다.