• BMB Reports
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• v.43 no.6
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• pp.427-431
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• 2010

Cyt2Aa2 is a mosquito-larvicidal protein produced as a 29 kDa crystalline protoxin from Bacillus thuringiensis subsp. darmstadiensis. To become an active toxin, proteolytic processing is required to remove amino acids from its N- and C-termini. This study aims to investigate the functional role of amino acid residues on the N-terminal ${\beta}1$ and C-terminal ${\alpha}F$ of Cyt2Aa2 protoxin. Mutant protoxins were constructed, characterized and compared to the wild type Cyt2Aa2. Protein expression data and SDS-PAGE analysis revealed that substitution at leucine-33 (L33) of ${\beta}1$ has a critical effect on dimer formation and structural stability against proteases. In addition, amino acids N230 and I233-F237 around the C-terminus ${\alpha}F$ demonstrated a crucial role in protecting the protoxin from proteolytic digestion. These results suggested that ${\beta}1$ and ${\alpha}F$ on the Nand C-terminal ends of Cyt2Aa2 protoxin play an important role in the molecular interaction and in maintaining the structural stability of the protoxin.

• Parasites, Hosts and Diseases
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• v.49 no.2
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• pp.145-151
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• 2011

The comparative efficacy of 2 anthelmintics (ivermectin and levamisole) against Baylisascaris transfuga migrating and encapsulated larvae was studied in mice. A total of 60 BALB/c mice inoculated each with about 1,000 embryonated B. transfuga eggs were equally divided into 6 groups (A-F) randomly. Mice of groups A and B were treated with ivermectin and levamisole, respectively, on day 3 post-infection (PI). Mice of groups A-C were killed on day 13 PI. Similarly, groups D and E were treated with ivermectin and levamisole, respectively, on day 14 PI, and all mice of groups D-F were treated on day 24 PI. The groups C and F were controls. Microexamination was conducted to count the larvae recovering from each mouse. The percentages of reduction in the number of migrating larvae recovered from group A (ivermectin) and B (levamisole) were 88.3% and 81.1%, respectively. In addition, the reduction in encapsulated larvae counts achieved by ivermectin (group D) and levamisole (group E) was 75.0% and 49.2%, respectively. The results suggested that, to a certain extent, both anthelmintics appeared to be more effective against migrating larvae than encapsulated larvae. However, in the incipient stage of infection, ivermectin may be more competent than levamisole as a larvicidal drug for B. transfuga.

• BMB Reports
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• v.34 no.4
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• pp.293-298
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• 2001

Tryptic activation of the 130-kDa Bacillus thuringiensis Cry4B $\delta$-endotoxin produced protease-resistant products of ca. 47 kDa and ca. 21 kDa. The 21-kDa fragment was identified as the N-terminal five-helix bundle (${\alpha}1-{\alpha}5$,) which is a potential candidate for membrane insertion and pore formation. In this study, we constructed the recombinant clone over-expressing this putative pore-forming (PPF) fragment as inclusion bodies in Escherichia coli. The partially purified inclusions were composed of a 23-kDa protein, which cross-reacted with Cry4B antibodies, and whose N-terminus was identical to that of the 130-kDa protein. Dissimilar to protoxin inclusions, the PPF inclusions were only soluble when the carbonate buffer, pH 9.0, was supplemented with 6 M urea. After renaturation via a stepwise dialysis, the refolded PPF protein appeared to exist as an oligomer and was structurally stable upon trypsin treatment. Unlike the 130kDa protoxin, the refolded protein was able to release entrapped glucose from liposomes, and showed comparable activity to the full-length activated toxin, although it lacks larvicidal activity These results, therefore, support the notion that the PPF fragment that consists of ${\alpha}1-{\alpha}5$ of the activated Cry4B toxin is involved in membrane pore-formation.

• The Korean Journal of Pesticide Science
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• v.11 no.1
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• pp.52-58
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• 2007

ALA (5-aminolevulinic acid) has been proposed as a tetrapyrrole-dependent photodynamic herbicide and insecticide by the action of the protoporphyrinogen IX oxidase (Protox IX). The present study was conducted to determine growth responses of plant and insects to ALA, biodegradable biopesticidal substance. In the paddy condition experiment, plant height and shoot fresh weight of barnyardgrass (Echinochloa crus-galli) was more reduced by ALA than rice plants, even though both plant species show great phytotoxicity. Hairy crabgrass (Digitaria sanguinalis), a monocot weed, was more sensitive to ALA at 5mM under upland condition when ALA applied on the foliage, compared with soybean (Glycine max) as a dicot crop. ALA solutions were tested for their insecticidal and larvicidal activities against Spodaptera exigua (Hubner) and Tetranychus urticae Koch. by foliar application and leaf-dipping method. The result showed higher insecticidal activity of ALA at 10mM and its mixture with insecticide luferon against S. exigua. Strongest insecticidal activity against T. urticae was observed from the ALA solution at 10mM 72 days after application. This results show that ALA solution had potent herbicidal and insecticidal activities against agricultural pests even though their activities were lower than those of synthetic pesticides.

• Korean journal of applied entomology
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• v.43 no.1
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• pp.35-41
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• 2004

Bacillus thuringiensis produces crystal proteins toxic to medically and agriculturally important pests during sporulation. To improve the activity of insecticidal crystal protein in applying to mosquito larval control, an expression vector, pSyn4D harboring the mosquitocidal cry11Aa gene under control of psbA promoter of Amaranthus hybridus was constructed. This expression vector was transformed into Synechocystis PCC6803 and a transformant, Tr2C was selected with kanamycin. The mosquitocidal cry11Aa gene was stably integrated Into genomic DNA of Tr2C in PCR detection using cry11Aa-specific primers. The transformant expressed 72-kDa Cry11Aa protein and median lethal time (LT$\sub$50/) was approximately 2.1 days for Culex tritaeniorhynchus larvae and 0.7 day for Anopheles sinensis larvae, respectively. These results suggest this transformant can be used for mosquito larval control as a biological control agent.

• BMB Reports
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• v.40 no.1
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• pp.58-64
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• 2007

Similar to the other known structures of Bacillus thuringiensis Cry $\delta$-endotoxins, the crystal structure of the 65-kDa activated Cry4Ba toxin comprises three domains which are, from the N- to C-terminus, a bundle of $\alpha$-helices, a three-$\beta$-sheet domain, and a $\beta$-sandwich. To investigate the properties of the C-terminal domain III in isolation from the rest of the toxin, the cloned Cry4Ba-domain III was over-expressed as a 21-kDa soluble protein in Escherichia coli, which cross-reacted with anti-Cry4Ba domain III monoclonal antibody. A highly-purified domain III was obtained in a monomeric form by ion-exchange and size-exclusion FPLC. Circular dichroism spectroscopy indicated that the isolated domain III fragment distinctly exists as a $\beta$-sheet structure, corresponding to the domain III structure embodied in the Cry4Ba crystal structure. In vitro binding analysis via immuno-histochemical assay revealed that the Cry4Ba-domain III protein was able to bind to the apical microvilli of the susceptible Stegomyia aegypti larval midguts, albeit at lower-binding activity when compared with the full-length active toxin. These results demonstrate for the first time that the C-terminal domain III of the Cry4Ba mosquito-larvicidal protein, which can be isolated as a native folded monomer, conceivably participates in toxin-receptor recognition.

• The Korean Journal of Pesticide Science
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• v.4 no.3
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• pp.75-81
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• 2000

These studies were carried out to investigate the toxicities of 38 registered insecticides to the sweetpotato whitefly (Trialeurodes vaporariorum). Insecticide activities were evaluated by testing systemic action and residual effect in the laboratory, and control efficacy in the greenhouse. All experiments were tested at the recommended concentration(ppm) of each insecticides. Insect growth regulators (IGRs), only pyriproxyfen showed over 90% of ovicidal effect. The insecticides that showed over 90% of larvicidal activity oil 3rd nymphal instars were abamectin, acetamiprid, chlorpyrifos-methyl, imidacloprid, pyripoxyfen, and acetamiprid+ethofenprox. Insecticides with 100% adulticidal activity were abamectin, acephate, acetamiprid, benfurcarb, bifenthrin, furathiocarb, endosulfan, fenitrothion, imidacloprid, phenthoate, pymetrozine, acetamiprid + ethofenprox, ethofenprox + diazinon, furathiocarb + difluberlzuron, and triazamate+${\alpha}$-cypermethrin. Abamectin, acetamiprid, imidacloprid, pyriproxyfen, and acetamiprid + ethofenpox showed both residual effect and systemic activity. In tile control efficacy test on B. tabaci, 90% control values were obtained at 11th day after treatment of the insecticides including abamectin, acetamiprid, imidacloprid, pyripoxyfen and acetamiprid + ethofenprox. These results indicate that abamectin, acetamiprid, imidacloprid, pyriproxyfen and acetamiprid + ethofenprox can be used for tile control of B. tabaci in field.

• Parasites, Hosts and Diseases
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• v.59 no.3
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• pp.189-225
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• 2021

The use of albendazole and mebendazole, i.e., benzimidazole broad-spectrum anthelmintics, in treatment of parasitic infections, as well as cancers, is briefly reviewed. These drugs are known to block the microtubule systems of parasites and mammalian cells leading to inhibition of glucose uptake and transport and finally cell death. Eventually they exhibit ovicidal, larvicidal, and vermicidal effects on parasites, and tumoricidal effects on hosts. Albendazole and mebendazole are most frequently prescribed for treatment of intestinal nematode infections (ascariasis, hookworm infections, trichuriasis, strongyloidiasis, and enterobiasis) and can also be used for intestinal tapeworm infections (taeniases and hymenolepiasis). However, these drugs also exhibit considerable therapeutic effects against tissue nematode/cestode infections (visceral, ocular, neural, and cutaneous larva migrans, anisakiasis, trichinosis, hepatic and intestinal capillariasis, angiostrongyliasis, gnathostomiasis, gongylonemiasis, thelaziasis, dracunculiasis, cerebral and subcutaneous cysticercosis, and echinococcosis). Albendazole is also used for treatment of filarial infections (lymphatic filariasis, onchocerciasis, loiasis, mansonellosis, and dirofilariasis) alone or in combination with other drugs, such as ivermectin or diethylcarbamazine. Albendazole was tried even for treatment of trematode (fascioliasis, clonorchiasis, opisthorchiasis, and intestinal fluke infections) and protozoan infections (giardiasis, vaginal trichomoniasis, cryptosporidiosis, and microsporidiosis). These drugs are generally safe with few side effects; however, when they are used for prolonged time (>14-28 days) or even only 1 time, liver toxicity and other side reactions may occur. In hookworms, Trichuris trichiura, possibly Ascaris lumbricoides, Wuchereria bancrofti, and Giardia sp., there are emerging issues of drug resistance. It is of particular note that albendazole and mebendazole have been repositioned as promising anti-cancer drugs. These drugs have been shown to be active in vitro and in vivo (animals) against liver, lung, ovary, prostate, colorectal, breast, head and neck cancers, and melanoma. Two clinical reports for albendazole and 2 case reports for mebendazole have revealed promising effects of these drugs in human patients having variable types of cancers. However, because of the toxicity of albendazole, for example, neutropenia due to myelosuppression, if high doses are used for a prolonged time, mebendazole is currently more popularly used than albendazole in anti-cancer clinical trials.

• International Journal of Industrial Entomology and Biomaterials
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• v.18 no.2
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• pp.105-112
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• 2009

The greenhouse whitefly, Trialeurodes vaporariorum (Westwood) and the sweet potato whitefly, Bemisia tabaci (Gennadius) are serious insect pests that have a wide host range including cucumber, tomato, and pepper. In this study, we tested larvicidal efficacy of several on-the-market environment-friendly agricultural materials (EFAM) to select the effective products after the target pests were stabilized in indoor rearing condition. The developmental periods of two whiteflies are as follows: in the case of T. vaporariorum, egg duration is 9.6 days and nymph is 18.9 days, and in the case of B. tabaci, egg duration is 7.4 days and nymph is 15.2 days under $25^{\circ}C$ with relative humidity (RH) of $60{\pm}5%$ and a photoperiod of 16 L:8D. The total period of T. vaporariorum was 5 days longer than B. tabaci. Among 22 EFAMs six products showed more than 60% of insecticide efficacy against T. vaporariorum BTV B, BTV D, BTV G, BTV L, BTV M, and BTV S. On the other hand, seven EFAM products showed over 60% of insecticide efficacy against B. tabaci BTV D, BTV G, BTV K, BTV L, BTV M, BTV N, and BTV U. In the case of Spodptera litura previously, 16 EFAMs were tested against $2^{nd}$ instar S. litura, and six EFAMs were found to have more than 90% efficacy. Test of these six EFAMs against entire larval stages were performed in this study. Although some of these products showed still more than 90% of insecticidal efficacy against up to $3^{rd}$ instar larvae, the efficacy of these EFAMs sharply decreased as ages increase, resulting in less than 60% of efficacy of the products at most. This result indicates the difficulty to control S. litura with the on-the-market EFAMs alone under economic injury level. Collectively, it is required to find more EFAMs, and find alternative method, and combined way of controlling to control those insect pests tested in this study.

• The Korean Journal of Pesticide Science
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• v.4 no.1
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• pp.51-58
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• 2000

These studies were carried out to investigate the toxicities of 43 registered insecticides to the sweetpotato whitefly(Bemisia tabaci, B. biotype). Insecticide activities were evaluated by testing systemic action and residual effect in the laboratory, and control efficacy in the greenhouse. All experiments were tested at the recommended concentration(ppm) of each insecticides. Insect growth regulators (IGRs), pyriproxyfen and teflubenzuron showed >95% ovicidal effect. The insecticides that showed >95% larvicidal activity on 3rd nymphal instars were abamectin, acetamiprid, imidacloprid, pyriproxyfen, and acetamiprid+ ethofenprox. Insecticides with >95% adulticidal activity were abamectin, acetamiprid, diazinon, endosulfan, fenitrothion, imidacloprid, methidathion, pirimiphos-methyl, pymetrozine, spinosad, acetamiprid+ ethofenprox, cartap kydrochloride+buprofezin, and fenpropathrin+fenitrothion. Abamectin, acetamiprid, imidacloprid, pyriproxyfen, and acetamiprid+ethofenprox showed both residual effect and systemic activity. In the control efficacy test on B. tabaci, 90% control values were obtained at 1st day after treatment of the insecticides including abamectin, acetamiprid, imidacloprid, pyriproxyfen and acetamiprid+ethofenprox but in pyriproxyfen, 90% control value was reached at 7th day after treatment. These results indicate that abamectin, acetamiprid, imidacloprid, pyriproxyfen and acetamiprid+ethofenprox can be used in control for B. tabaci in field.