Purification and partial characterization of a glutathione S-transferase from the two-spotted spider mite, Tetranychus urticae

Glutathione S-transferases (GSTs) are known to catalyze conjugations by facilitating the nucleophilic attack of the sulfhydryl group of endogenous reduced glutathione on electrophilic centers of a vast range of xenobiotic compounds, including insecticides and acaricides. Elevated levels of GSTs in the two-spotted spider mite, Tetranychus urticae Koch, have recently been associated with resistance to acaricides such as abamectin [Pestic. Biochem. Physiol. 72 (2002) 111]. GSTs from acaricide susceptible and resistant strains of T. urticae were purified by glutathione-agarose affinity chromatography and characterized by their Michaelis-Menten kinetics towards artificial substrates, i.e., 1-chloro-2,4-dinitrobenzene and monochlorobimane. The inhibitory potential of azocyclotin, dicumarol, and plumbagin was low (IC₅₀ values > 100 μM), whereas ethacrynic acid was much more effective, exhibiting an IC₅₀ value of 4.5 μM. GST activity is highest in 2-4-day-old female adults and dropped considerably with progressing age. Furthermore, molecular characteristics were determined for the first time of a GST from T. urticae, such as molecular weight (SDS-PAGE) and N-terminal amino acid sequencing (Edman degradation). Glutathione-agarose affinity purified GST from T. urticae strain WI has a molecular weight of 22.1 kDa. N-terminal amino acid sequencing revealed a homogeneity of ≈50% to insect GSTs closely related to insect class I GSTs (similar to mammalian Delta class GSTs).     

芒果果实冷害过程中谷胱甘肽和抗坏血酸含量的变化

‘紫花’芒果果实（ＭａｎｇｉｆｅｒａｉｎｄｉｃａＬ．ＺｈｉＨｕａ）２℃贮藏１５ｄ时发生严重的冷害。贮藏前期,还原型谷胱甘肽（ＧＳＨ）、还原型维生素Ｃ（ＡｓＡ）含量略有上升,说明短期低温胁迫可促使机体提高清除氧自由基能力,以适应不良环境。随冷害时间延长,ＧＳＨ、ＡｓＡ含量下降;膜脂过氧化产物（ＭＤＡ）、膜透性、氧化型谷胱甘肽（ＧＳＳＧ）和氧化型维生素Ｃ（ＤＨＡ）增加,表明细胞防御能力下降,自由基代谢平衡可能被打破,膜脂过氧化加剧,果实冷害加重。１０℃贮藏的芒果,无冷害症状,２２ｄ时果实启动成熟,贮藏期间ＧＳＨ、ＡｓＡ含量维持较高水平。     

钙处理对红星苹果H2O2含量和GSH—PX活性的影响

红星苹果室温贮藏时,硬度下降;Ｈ２Ｏ２含量先下降,后升高至峰值,末期下降;过氧化氢酶和谷胱甘肽过氧化物酶活性分别呈双峰和单峰曲线。渗钙处理能延缓果肉软化,降低前期Ｈ２Ｏ２含量,提高谷胱甘肽过氧化物酶活性,对过氧化氢酶活性的影响较小。     

外源水杨酸对黄瓜叶片几种酶活性和抗氧化物质含量的影响

 采用不同浓度的水杨酸(SA) 对营养液培养的黄瓜( Cucumis sativus L. ) 进行处理, 结果表明, 不同浓度的SA 均明显提高了黄瓜叶片中过氧化物酶、多酚氧化酶、苯丙氨酸解氨酶的活性, 其中以SA 200μmol·L^-1处理的效果最为显著。SA 100μmol·L^-1的处理显著抑制了IAA 氧化酶和抗坏血酸氧化酶的活性, 而SA 200 和400μmol·L^-1的处理则有促进作用。SA 100μmol·L^-1的处理显著增加了黄瓜叶片中还原型抗坏血酸、还原型谷胱甘肽和氧化型谷胱甘肽的含量。     

富硒茶对大鼠抗氧化功能的影响

用基础低硒饲料饲养大鼠 ,以自来水 (对照 )、Na2 SeO3 溶液、低硒茶水和富硒茶水作试验处理 ,实验 7周后分析血液及肝脏中谷胱甘肽过氧化物酶 (GSH Px)活性、超氧化物歧化酶 (SOD)活性和血液、肝脏的含硒量 ,丙二醛 (MDA)的含量。结果表明 :富硒茶显著增加大鼠血液和肝脏的含硒量 ,降低MDA含量 ,提高血液的SOD活性。富硒茶处理大鼠血液的GSH Px活性是对照和低硒茶叶的 1 3倍 ,肝脏GSH Px活性可达 1 4倍 ,富硒茶在提高肝脏和血液的GSH Px活性和SOD活性 ,降     

Hepatoprotective effect of Ginkgoselect Phytosome in rifampicin induced liver injury in rats: evidence of antioxidant activity

The protective effects of Ginkgoselect Phytosome((R)) (GBP) on Rifampicin (RMP) induced hepatotoxicity and the probable mechanism(s) involved in this protection were investigated in rats. Liver damage was induced in Wistar rats by administering rifampicin (500 mg/kg, p.o.) daily for 30 days. Simultaneously, GBP at 25 mg/kg and 50 mg/kg, and the reference drug silymarin (100 mg/kg) were administered orally for 30 days/daily to RMP treated rats. Levels of marker enzymes (SGOT, SGPT and SALP), albaumin (Alb) and total proteins (TP) were assessed in serum. The effects of GBP on lipid peroxidation (LPO), reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GR) were assayed in liver homogenates to evaluate antioxidant activity. GBP (25 and 50 mg/kg) and silymarin elicited a significant hepatoprotective activity by lowering the levels of serum marker enzymes and lipid peroxidation and elevated the levels of GSH, SOD, CAT, GPX, GR, Alb and TP in a dose dependant manner. The present findings suggest that the hepatoprotective effect of GBP in RMP induced oxidative damage may be related to its antioxidant and free radical scavenging activity.     

Resistance to chlorpyrifos and lambda-cyhalothrin along with detoxifying enzyme activities in field-collected female populations of European red mite

The levels of susceptibility of populations of the European red mite Panonychus ulmi (Koch) (Acarina: Tetranychidae) collected from apple orchards in the Bursa region of Turkey to the insecticides chlorpyrifos and lambda-cyhalothrin, were determined by a petri leaf disk—Potter spray tower method. When compared with the susceptible population, resistance ratios at the LC₅₀ level ranged from 6.0- to 35.6-fold, and from 0.7- to 5.7-fold for chlorpyrifos and lambda-cyhalothrin, respectively. Kinetic parameters of general esterase activity with α-naphthyl acetate as substrate indicated that an increased activity was present in the resistant populations compared with the susceptible populations. In these strains, 1.5- and 2.2-fold higher Glutathione S-transferase (GST) activity was also detected with the substrate 1-chloro-2,4-dinitrobenzene. General esterase activity gel profiles of these populations were studied by native polyacrylamide gel electrophoresis, but no relationship between resistance ratios and band patterns was detected. The results of this study document a decreased efficacy of chlorpyrifos and lambda-cyhalothrin in field populations of P. ulmi in Turkey, possibly linked to altered activities of esterases and GST.     

贮前外源水杨酸处理对桃果冷贮性的影响

选择商品成熟期的桃(Prunus persica L. cv. Beijing 24),采摘后分别用0、0.35、0.7、1.0 mmol/L 水杨酸 （Salicylic acid,SA）溶液浸泡5 min,在0℃的低温下贮藏28d,然后转到20℃下3d（模拟货架期）。货架期结束时,观察果实冷害和腐烂发生状况并计算果实冷害指数和腐烂指数,测定果实的硬度和丙二醛含量。结果表明,与对照相比,1 mmol/L SA处理对维持果实硬度具有显著的效果,同时显著降低了冷害指数、腐烂指数和丙二醛含量,其他两种浓度的SA处理与对照之间不存在显著差异。说明1 mmol/L SA能够减轻桃果的冷害。进一步研究发现,在冷藏中期（14d）、冷藏结束时（28d）和货架期结束时,1 mmol/L SA预处理的桃果肉的还原性抗坏血酸与氧化性抗坏血酸含量之比(AsA/DHAsA)分别比对照高39%,61%和55%。在冷藏中期（14d）,还原性谷胱甘肽与氧化性谷胱甘肽含量之比(GSH/GSSG) 比对照高68%。在贮藏和货架期结束时,抗坏血酸氧化酶和谷胱甘肽还原酶活性显著高于对照。上述结果说明,水杨酸能减轻桃果由于冷贮发生的冷害与诱导提高抗氧化系统抗抗坏血酸—谷胱甘肽循环的活性有关,适宜浓度的水杨酸处理可能是提高桃果实冷藏性的有效措施。     

Human metabolism of atrazine

Atrazine (ATZ) metabolism by human liver microsomes (HLM), cytochrome P450 (CYP) isoforms, and human liver (HL) S9 fractions, was investigated using HPLC/PDA and LC/MS/MS. CYP-dependent metabolites from pooled HLM are desethylatrazine (DEA), desisopropylatrazine (DIA), 1-hydroxyisopropylatrazine (HIATZ), and 2-hydroxyethyl atrazine (HEATZ). DEA and DIA were major metabolites in pooled HLM. CYP1A2 and 2C19, respectively, were major isoforms for DEA and DIA production. CYP3A4, while less active, is generally at high concentrations, produces both DEA and DIA and is significant. The percent total normalized rates (%TNR) for CYP1A2 and 3A4 in pooled HLM were 63% and 24% for DEA, and 35% and 56% for DIA production. Single donor HLM samples, showed correlations for CYP1A2 (r = 0.92) and 3A4 (r = 0.81) for DEA and DIA production, while variations in production of DEA and DIA were 8.5- and 6.0-fold, respectively. Pooled S9 fractions also mediate glutathione conjugation of atrazine, DEA and DIA.     

Resistance in Maconellicoccus hirsutus (Green) in India to selected insecticides and quantification of detoxifying enzymes imparting resistance

Pink mealybug Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae) is a destructive pest of agricultural and horticultural crops. Insecticides are the major tool used to control M. hirsutus. The present study was conducted to evaluate resistance to the commonly used insecticides acephate, dichlorvos, imidacloprid and buprofezin in M. hirsutus collected from seven different geographical locations of mulberry and vineyards in India. Detoxifying enzymes, namely esterase, glutathione S transferase (GST) and cytochrome P-450 (cyt-P450), were quantified in populations. One population from Erode showed a low level of resistance to acephate (resistance ratio [RR] 10.3-fold), one from Salem showed a low level of resistance to dichlorvos (resistance ratio [RR] 13.7-fold), one from Sangli showed a very low level of resistance to imidacloprid (RR 10.2-fold), and one from Chikkaballapur showed a low level of resistance to buprofezin (RR 14.8-fold). Activity ratios for detoxifying enzymes ranged from 1.8- to 4.9-fold for GST, 1.8- to 3.7-fold for esterase and 1.9- to 2.4-fold for cyt-P450. Furthermore, organophosphate resistance and activity of enzymes (esterase, GST and cyt-P450) were positively correlated. To contain the evolution of resistance to M. hirsutus infestation, buprofezin and imidacloprid could be used, supplemented with biointensive integrated management strategies and regular resistance monitoring programs.