Regulation of JH epoxide hydrolase versus JH esterase activity in the cabbage looper, Trichoplusia ni, by juvenile hormone and xenobiotics

JH III esterase and JH III epoxide hydrolase (EH) in vitro activity was compared in whole body Trichoplusia ni homogenates at each stage of development (egg, larva, pupa and adult). While activity of both enzymes was detected at all ages tested, JH esterase was significantly higher than EH activity except for day three of the fifth (last) stadium (L5D3). For both enzymes, activity was highest in eggs. Adult virgin females had 4.6- and 4.0-fold higher JH esterase and EH activities, respectively, than adult virgin males. JH III metabolic activity also was measured in whole body homogenates of fifth stadium T. ni that were fed a nutritive diet (control) or starved on a non-nutritive diet of alphacel, agar and water. With larvae that were starved for 6, 28 and 52 h, EH activity per insect equivalent was 48%, 5% and 1%, respectively, of the control insects. At the same time points, JH esterase activity levels in starved T. ni were 29%, 4% and 3% of that of insects fed the nutritive diet. Selected insect hormones and xenobiotics were administered topically or orally to fifth stadium larvae for up to 52 h, and the effects on whole body EH and JH esterase activity analyzed. JH III increased the JH III esterase activity as high as 2.2-fold, but not the JH III EH activity. The JH analog, methoprene, increased both JH esterase and EH activity as high as 2.5-fold. The JH esterase inhibitor, 3-octylthio-1,1,1-trifluoropropan-2-one (OTFP), had no impact on EH activity. The epoxides trans- and cis-stilbene oxide (TSO and CSO) in separate experiments increased the EH activity approximately 2.0-fold. TSO did not alter JH esterase levels when topically applied, but oral administration reduced activity to 70% of the control at 28 h, and then increased the activity 1.8-fold at 52 h after the beginning of treatment. CSO had no effect on JH esterase activity. Phenobarbital increased EH activity by 1.9-fold, but did not change JH esterase levels. Clofibrate and cholesterol 5alpha,6alpha-epoxide had no effect on EH. JH esterase activity also was not affected by clofibrate, but cholesterol 5alpha,6alpha-epoxide reduced the JH esterase activity to 60-80% of the control. The biological significance of these results is discussed.     

Decapitation impacting effect of topically applied chlorpyrifos on acetylcholinesterase and general esterases in susceptible and resistant German cockroaches (Dictyoptera: Blattellidae)

The effect of topically applied chlorpyrifos on acetylcholinesterase and other esterases in heads and decapitated bodies of CSMA and Crawford German cockroaches was examined with spectrophotometric enzyme assay and native polyacrylamide gel electrophoresis. The toxicity of chlorpyrifos was greatly reduced in decapitated CSMA male cockroaches with LD50 value 17.1-fold higher than that of normal CSMA cockroaches. Acetylcholinesterase activity from heads was significantly higher in the Crawford compared with the CSMA strain and did not change until 24 h after chlorpyrifos in vivo treatment in both strains. The p-nitrophenyl butyrate (NPB) esterase activities from both heads and decapitated bodies of the resistant Crawford strain were significantly greater than the susceptible CSMA strain. The p-NPB esterase activity was significantly inhibited by chlorpyrifos in vivo treatment, and total p-NPB esterase activity was significantly reduced in decapitated bodies compared with heads of both strains. Native polyacrylamide gel electrophoresis (PAGE) analysis of extracts solubilized with Triton X-100 from heads and decapitated bodies revealed five major esterase bands and an acetylcholinesterase (AChE) band with a high capability of hydrolyzing alpha-naphthyl butyrate and acetylthiocholine, respectively. In the heads of susceptible CSMA male cockroaches, the activity of mobile isozymes d1 and d2 was completely inhibited at 24 h after chlorpyrifos application, and isozyme e was partially inhibited. In contrast, isozymes c1 and c2 from the decapitated bodies of CSMA cockroaches were mostly affected at 24 h after the topical application of chlorpyrifos. The activities of acetylcholinesterase and esterase isozymes a and b from the decapitated body remained uninhibited in both strains. Inhibition of isozymes d1 and d2 seems to be more important in chlorpyrifos intoxication than acetylcholinesterase.     

An esterase is involved in geraniol production during palmarosa inflorescence development

Total incorporation of exogenously administered [2-14C]acetate into essential oil of palmarosa (Cymbopogon martinii) was found to be relatively higher than that of either [U-14C]sucrose or [U-14C]glucose during inflorescence development. Among the major essential oil constituents, biogenesis of geranyl acetate was much higher than that of geraniol. Alkaline hydrolysis of [14C]labeled geranyl acetate revealed that the majority of the label incorporated into geranyl acetate was present in the geraniol moiety, indicating that only newly synthesized geraniol gets acetylated to form geranyl acetate. Geranyl acetate cleaving esterase (GAE) activity followed a similar pattern during both in vivo and in vitro inflorescence development, with maximum activity at immature inflorescence stages, suggesting the involvement of GAE in geraniol production during inflorescence development. Five esterase isozymes (Est-A to E) were detected in the enzymic fraction of palmarosa inflorescence and all showed GAE activity, with Est-B being significantly increased during inflorescence development. The role of GAE in geraniol production and improving the palmarosa oil quality is discussed.     

Effects of emulsified perfluorochemicals on liver aryl esterase in rats.

1. The effects of intravenous (i.v.) injection of various perfluorochemical (PFC) emulsions have been studied separately in male and female rats. 2. Injection of 10 ml/kg body weight of either Fluosol-DA 20% (F-DA) or a novel perfluorodecalin emulsion containing a C-16 oil additive in male rats increased liver weight up to 7 days later; no corresponding change occurred in response to injection of Oxypherol (FC-43). 3. Liver weight was also increased in female rats at 72 hr after injection of the novel emulsion but this was less pronounced than in males; liver weight in female rats was unchanged in response to injection of either F-DA or FC-43. 4. Mean liver aryl esterase activity in male rats was increased 2- to 3-fold (P less than 0.05) at 7 days after injection of the novel emulsion. No significant alterations in aryl esterase activity occurred in response to injection of either F-DA or FC-43, although in both cases there was a trend towards increased activity. 5. Liver aryl esterase activity in female rats was significantly (P less than 0.05) decreased at 72 hr following FC-43 injection with similar, but much less pronounced, changes occurring in response to injection of F-DA and the novel emulsion. 6. These results show that injection of a single low dose of emulsified PFCs into rats can alter hepatic microsomal aryl esterase activity but the response is highly variable, depending on composition of emulsion injected and sex of recipient.     

Effect of 5-thio-D-glucose on testosterone biosynthesis in vivo in mice testes

Testicular synthesis of (14C)cholesterol and (14C)testosterone from (14C)acetate were investigated in mice treated with 5-thio-D-glucose at a dose of 33 mg/kg body weight/day for 21 days. The testicular synthesis of free cholesterol as well as steroids were significantly decreased. The steroid synthesizing enzymes, cholesterol esterase, cholesterol side-chain cleaving enzyme, total alpha-hydroxysteroid dehydrogenase and total beta-hydroxysteroid dehydrogenase, were also analysed. Cholesterol esterase and total beta-hydroxysteroid dehydrogenase were significantly reduced whereas total alpha-hydroxysteroid dehydrogenase was unaffected. Hence, a decrease in free cholesterol for steroid synthesis and a decreased activity of the steroidogenic enzyme, beta-hydroxysteroid dehydrogenase, were responsible for the diminished synthesis of testosterone.     

艾的挥发性物质化感作用研究

艾是我国常见的菊科多年生杂草,也是草本药用植物。研究表明,艾的茎叶在密闭系统中产生的挥发性物质对稗草、三叶鬼针草和青葙的幼苗生长有显著抑制作用。稗草的根长、苗高和鲜重分别比对照减少43.3%、30.8%和35.7%,三叶鬼针草分别比对照减少29.2%、30.0%和29.5%,青葙分别比对照减少26.3%、15.4%和27.9%。水蒸汽蒸馏法所得的挥发油对稗草、三叶鬼针草和青葙的幼苗生长也有显著抑制作用。稗草的根长、苗高和鲜重分别比对照减少71.5%、54.8%和30.5%,三叶鬼针草根长、苗高和鲜重分别比对照减少40.9%、49.2%和39.1%,青葙根长、苗高和鲜重分别比对照减少78.8%、29.1%和34.1%。结果显示,艾挥发物对邻近植物可能产生化感作用。     

Homeostatic regulation of acetazolamide-sensitive esterase activity in the bluegill sunfish, Lepomis macrochirus, following acute cold shock

Acetazolamide-sensitive esterase activity was elevated in branchial homogenates of control juvenile bluegill sunfish, Lepomis macrochirus , acclimated at 20° C but decreased rapidly within 9 h following an acute hypothermal shock to 8° C. After 2 weeks at 8° C, shocked-fish enzyme activity was similar to control fish. At 20° C acclimation temperature, specific activity of bluegills was similar in swimbladder, liver, kidney, gill, spleen, and gonad homogenates and was significantly higher (α=0.05 level) in whole blood homogenates. The pH optima for enzymes extracted from fish acclimated at 20° and 8° C were 7.29 and 8.00, respectively. Polyacrylamide gel electrophoresis (PAGE) demonstrated two distinct forms of acetazolamide-sensitive esterase activity present in both 20° and 8° C acclimated fish. Specific activity for homogenates from both 20° and 8° C acclimated fish differed significantly when assayed at 20° C, suggesting both qualitative and quantitative changes in acetazolamide-sensitive esterase. It is postulated that relatively rapid alterations in esterase activity promote survival in bluegill following acute cold shock through the central role of enzymes in the regulation of plasma ion concentrations and acid/base equilibria.     

Purification and properties of extracellular lipase from Pseudomonas aeruginosa EF2.

Extracellular lipase was purified from a Tween 80-limited continuous culture of Pseudomonas aeruginosa EF2 by ultrafiltration of the culture supernatant followed by anion-exchange and gel-filtration FPLC. The lipase was composed of a single subunit (Mr 29,000, pI 4.9), which was capable of a variable degree of aggregation, and which exhibited both lipase activity, measured with the insoluble substrate olive oil (predominantly triolein), and esterase activity, measured with the soluble substrates p-nitrophenyl acetate and Tween 80. Lipase activity was approximately eight times higher than either type of esterase activity (kcat approximately 3000 s-1 for the hydrolysis of olive oil). The enzyme showed a marked regiospecificity for the 1,3-oleyl residues of radiolabelled triolein, was relatively stable at moderate temperatures (exhibiting a biphasic loss of activity with an initial t1/2 of 17.5 min at 60 degrees C) and was very stable to freezing and thawing. Lipase activity was only weakly inhibited by the serine-active reagent 3,4-dichloroisocoumarin, and was not inhibited by the chelating agent EDTA (1 mM). The N-terminal amino acid sequence of the Ps. aeruginosa EF2 lipase showed a marked similarity to those of several other bacterial lipases.     

Effect of Satureja hortensis L. essential oil on feeding efficiency and biochemical properties of Glyphodes pyloalis Walker (Lepidoptera: Pyralidae)

The effect of essential oil of Satureja hortensis L. on food consumption, biochemical compounds and digestive and detoxification enzymes activity of lesser mulberry pyralid, Glyphodes pyloalis Walker was investigated. The percentage of feeding inhibition was dose dependent. The S. hortensis essential oil exhibited a significant antifeedant activity at the highest concentration used. The feeding indices such as efficiency of conversion of ingested food (ECI), efficiency of conversion of digested food (ECD), relative growth rate (RGR) and relative consumption rate (RCR) were decreased compared with the control in fifth instar larvae, while approximate digestibility (AD) index increased significantly. Results showed significant decreases in the amount of protein, lipid, carbohydrates and the activity level of α-amylase enzyme in treated larvae with S. hortensis essential oil. The activity level of detoxifying enzymes such as esterase and glutathione S-transferase were significantly affected by S. hortensis essential oil. The results of this study show high deterrence and antifeedant activity of this essential oil which deserves consideration in integrated pest management program.     

Extracellular carboxylesterase activity of Fusarium graminearum

Summary The extracellular carboxylesterase produced by Fusarium graminearum was investigated. The esterase showed optimum activity at 37°C and at pH 8.7–9.0. Increased activity was observed when whey as a growth substrate was supplemented with either Tween 40 or olive oil. Good activity was also obtained when the fungus was grown on a semi-solid bran culture medium. The esterase showed increased activity towards esters containing short-chain fatty acids.     

Seasonal differences in methyl farnesoate esterase activity in tissues of the spider crab Libinia emarginata

Summary

Methyl farnesoate (MF), an unepoxidated form of insect JH III, is present in Crustacea. MF is synthesized by the mandibular organs and is degraded to fomesoic acid (FA) by peripheral tissues. In this study we investigated MF degradation by esterases in hepatopancreas, ovary, testes and hemolymph of the spider crab Libinia emarginata collected at different times of the year to determine seasonal differences. The conversion of MF to FA varied among the tissues. In the summer, the hepatopancreas showed the greatest esterase activity (52.8% conversion in females and 59.16% in males), and it was twice as high (28.86%) in ovaries than in the testes (12.16%), but was low in the hemolymph of both sexes (10.84% in males, and 6.97% in females). In the fall, the conversion of MF to FA was significantly reduced in all tissues (ovary 8.55%, testes 6.21%, hepatopancreas 10.22%, hemolymph 3.96%). Eyestalk ablation of animals in the fall restored MF esterase activity to summer levels. When tissues from these animals were incubated with OTFP, a specific inhibitor of JH esterase, MF metabolism was significantly reduced. These results suggest that MF esterase activity depends on direct induction by MF, and its degradation is by a specific esterase(s).     

Biotransformation of geranyl acetate to geraniol during palmarosa (Cymbopogon martinii, Roxb. wats. var. motia) inflorescence development

Only immature palmarosa (Cymbopogon martinii, Roxb. wats. var. motia) inflorescence with unopened spikelets accumulated essential oil substantially. Geraniol and geranyl acetate together constituted about 90% of the palmarosa oil. The proportion of geranyl acetate in the oil decreased significantly with a corresponding increase of geraniol, during inflorescence development. An esterase enzyme activity, involved in the transformation of geranyl acetate to geraniol, was detected from the immature inflorescence using a gas chromatographic procedure. The enzyme, termed as geranyl acetate cleaving esterase (GAE), was found to be active in the alkaline pH range with the optimum at pH 8.5. The catalysis of geranyl acetate was linear up to 6 h, and after 24 h of incubation, 75% of the geranyl acetate incubated was hydrolyzed. The GAE enzymic preparation, when stored at 4 degrees C for a week, was quite stable with only 40% loss of activity. The physiological role of GAE in the production of geraniol during palmarosa inflorescence development has been discussed.     

Alterations in esterases are associated with malathion resistance in Habrobracon hebetor (Hymenoptera: Braconidae)

Biochemical mechanisms of malathion resistance were investigated in a malathion-resistant strain of the parasitoid Habrobracon hebetor Say collected from a farm storage in Kansas. General esterase activities were significantly lower in the resistant strain compared with those in a susceptible strain. However, no significant differences were found in activities of malathion specific carboxylesterase (MCE), glutathione S-transferase and cytochrome P450 dependent O-demethylase activities, cytochrome P450 contents, and sensitivity of acetylcholinesterase to inhibition by malaoxon between the 2 strains. Because MCE was not elevated in the resistant strain, the weak malathion resistance in H. hebetor may result from a different mechanism compared with that hypothesized for some insect species in which reduced general esterase activity is accompanied by an elevated MCE. Decreased esterase activity in the resistant strain suggested that null alleles of some esterases were associated with the resistance. Indeed, E1 and E2, major esterases in the susceptible strain, were not present in the resistant strain on polyacrylamide gels that were stained for esterase activity using the model substrate 1-naphthyl acetate. In contrast, the activity of esterase E3 on the gels was much higher in the resistant strain as compared with that of the susceptible strain. These findings indicate that malathion resistance in H. hebetor is associated with both an increased activity of the esterase E3 and null alleles of the esterases E1 and E2.     

Elevated esterase activity in resistant tufted apple bud moth, Platynota idaeusalis (Walker) (Lepidoptera: Tortricidae).

1. Esterase activity in the tufted apple bud moth was measured spectrophotometrically by the hydrolysis of alpha-naphthyl acetate. 2. Resistant populations from both laboratory and field exhibited significantly greater esterase activity than did the respective susceptible populations. 3. The resistant laboratory population had significantly higher esterase activity than the resistant field population, suggesting that this field population contained a mixture of resistant and susceptible individuals.     

反枝苋水浸提液与挥发油对黄瓜根尖的影响

采用悬空气法研究了在入侵植物反枝苋(Amaranthus retroflexus L.)水浸提液和挥发油作用下,黄瓜根缘细胞活性、根冠果胶甲基酯酶(PME)、根尖过氧化氢酶(CAT)、过氧化物酶(POD)、超氧化物歧化酶(SOD)以及丙二醛(MDA)含量的变化规律.结果表明:反枝苋水浸提液对黄瓜根的生长无显著性影响而挥发油显著抑制黄瓜根的生长,且随浓度增大抑制作用显著增强.PME活性随着水浸提液浓度的增大呈先上升后下降趋势,而随着挥发油浓度的升高呈现逐渐上升的趋势;水浸提液和挥发油均降低了对根缘细胞的存活率,这种抑制作用随浓度的增加而增大;随着处理液浓度增大,黄瓜根尖中MDA含量、CAT活性整体表现为增加,SOD活性先升高后降低,POD活性与对照差异不显著.反枝苋挥发油的化感效应大于水浸提液的化感效应.     

Virucidal effect of peppermint oil on the enveloped viruses herpes simplex virus type 1 and type 2 in vitro

The virucidal effect of peppermint oil, the essential oil of Mentha piperita, against herpes simplex virus was examined. The inhibitory activity against herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) was tested in vitro on RC-37 cells using a plaque reduction assay. The 50% inhibitory concentration (IC50) of peppermint oil for herpes simplex virus plaque formation was determined at 0.002% and 0.0008% for HSV-1 and HSV-2, respectively. Peppermint oil exhibited high levels of virucidal activity against HSV-1 and HSV-2 in viral suspension tests. At noncytotoxic concentrations of the oil, plaque formation was significantly reduced by 82% and 92% for HSV-1 and HSV-2, respectively. Higher concentrations of peppermint oil reduced viral titers of both herpesviruses by more than 90%. A clearly time-dependent activity could be demonstrated, after 3 h of incubation of herpes simplex virus with peppermint oil an antiviral activity of about 99% could be demonstrated. In order to determine the mode of antiviral action of the essential oil, peppermint oil was added at different times to the cells or viruses during infection. Both herpesviruses were significantly inhibited when herpes simplex virus was pretreated with the essential oil prior to adsorption. These results indicate that peppermint oil affected the virus before adsorption, but not after penetration into the host cell. Thus this essential oil is capable to exert a direct virucidal effect on HSV. Peppermint oil is also active against an acyclovir resistant strain of HSV-1 (HSV-1-ACV(res)), plaque formation was significantly reduced by 99%. Considering the lipophilic nature of the oil which enables it to penetrate the skin, peppermint oil might be suitable for topical therapeutic use as virucidal agent in recurrent herpes infection.     

Susceptibility and detoxifying enzyme activity in two spider mite species (Acari: Tetranychidae) after selection with three insecticides

Changes in the susceptibility and detoxifying enzyme activity were measured in laboratory strains of Banks grass mite, Oligonychus pratensis (Banks), and twospotted spider mite, Tetranychus urticae Koch, that were repeatedly exposed to three insecticides. Three strains of each mite species were exposed to one of two pyrethroids, bifenthrin, and lambda-cyhalothrin, or an organophosphate, dimethoate, for 10 selection cycles at the LC60 for each insecticide. A reference or nonselected strain of each mite species was not exposed to insecticides. After 10 cycles of exposure, susceptibility to the corresponding insecticides, bifenthrin, lambda-cyhalothrin, and dimethoate, decreased 4.5-, 5.9-, and 289.2-fold, respectively, relative to the reference strain in the respective O. pratensis strains, and 14.8-, 5.7-, and 104.7-fold, respectively, relative to the reference strain in the respective T. urticae strains. In the bifenthrin-exposed O. pratensis strain, there was a 88.9-fold cross-resistance to dimethoate. In the dimethoate-exposed T. urticae strain, there was a 15.9-fold cross-resistance to bifenthrin. These results suggest that there may be cross-resistance between dimethoate and bifenthrin. The reduced susceptibility to dimethoate remained stable for three months in the absence of selection pressure in both mites. The decrease in susceptibility in the O. pratensis strains exposed to bifenthrin, lambda-cyhalothrin, and dimethoate was associated with a 4.7-, 3.0-, and 3.6-fold increase in general esterase activity, respectively. The decrease in susceptibility in the T. urticae strains exposed to bifenthrin and lambda-cyhalothrin was associated with a 1.3- and 1.1-fold increase in general esterase activity, respectively. The mean general esterase activity was significantly higher in the pyrethroid-exposed O. pratensis and T. urticae strains than in the nonselected strain. There was no significant increase in esterase activity in the dimethoate-exposed T. urticae strain. The decrease in susceptibility to insecticides was also associated with reduced glutathione S-transferase 1-chloro-2, 4-dinitrobenzene conjugation activity, but this did not appear to be related to changes in insecticide susceptibility. These results suggest that in these mites, the general esterases may play a role in conferring resistance to pyrethroids. However, some other untested mechanism, such as target site insensitivity, must be involved in conferring dimethoate resistance.     

桉树精油及其活性物质对干鱼害虫白腹皮蠹的熏蒸毒性

为了评估桉树精油和α-蒎烯对白腹皮蠹Dermestes maculatus Degeer的杀虫作用,采用生测方法测定了不同剂量和不同处理时间下,其幼虫和成虫的死亡率。结果表明:用桉树精油及其活性成分α-蒎烯处理白腹皮蠹成虫和幼虫后,可显著影响其死亡率（P＜0.001）。白腹皮蠹的反应因化学物种类、剂量和暴露时间不同而异。当剂量为32 μL/cm³的桉树精油处理白腹皮蠹幼虫72 h后,死亡率可超过90%。同样的剂量对其成虫熏蒸72 h后也表现高毒。在相同的剂量和相同的暴露时间下,α-蒎烯对白腹皮蠹的成虫和幼虫也表现高毒,且成虫的抗性比幼虫强。Probit检验结果表明桉树精油比α-蒎烯更有效。以上结果说明这些天然产物对白腹皮蠹种群的控制是很有用的。     

Lipase of Mucor pusillus

Lipase of Mucor pusillus NRRL 2543 was recovered with ammonium sulfate precipitation, gel filtration on Sephadex G-75, and anion-exchange chromatography on diethylaminoethyl-Sephadex A-50. Maximal glycerol ester hydrolase (lipase) activity was observed at pH 5.0 to 5.5 and 50 C when trioctanoin and olive oil were used as substrates. The enzyme also showed esterase activity; it hydrolyzed, with the exception of methyl butyrate, all methyl esters tested. A minimum chain length of six carbons appeared to be a requirement for esterase activity, which was maximal at about pH 5.5 with methyl dodecanoate (C(12)) as the substrate. Neither the glycerol ester hydrolase (lipase) nor the esterase activity of the enzyme appeared to be affected by thiol group inhibitors, chelating agents, and reducing compounds. On the other hand, hydrolysis of triolein and methyl dodecanoate was arrested to the same extent in the presence of diisopropyl fluorophosphate, which suggested the involvement of serine in the active center of the enzyme. The enzyme remained stable during a 30-day storage at - 10 C.     

ON THE ASSOCIATION OF NON-SPECIFIC ESTERASE ACTIVITY WITH CENTRAL NERVE MYELIN PREPARATIONS

Abstract— Isolated bovine central nerve myelin sheath preparations showed non-specific esterase activity towards naphthyl ester substrates of increasing chain length from acetate to palmitate. Short chain esters were hydrolysed much faster than long chain substrates by myelin, the specific activity for the hydrolysis of β-naphthyl acetate being the highest. Micro-somal fractions from brain white matter were much higher in esterase activity to all naphthyl ester substrates. NADPH-cytochrome c reductase activity was absent from isolated myelin samples. Distilled water and salt and buffer solutions of different ionic strengths and pH were ineffective in releasing non-specific esterase activity from myelin although tri-potassium citrate caused marked inhibition of the membrane-bound esterase activity. The detergent Triton X-100 released esterase activity from the myelin preparations but at a concentration of 0.1 per cent was also inhibitory.