• 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.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.21 no.4
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• pp.289-296
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• 2018

Purpose: We aimed to study the pattern of liver-injury in children with dengue fever (DF) and validate serum aminotransferase ${\geq}1,000IU/L$ as a marker of severe DF. Methods: Children admitted with DF were included. DF was defined by presence of clinical criteria and positive serological or antigen tests in absence of other etiology. DF severity was graded as dengue without or with warning signs and severe dengue. Liver-injury was defined as alanine aminotransferase (ALT) more than twice the upper limit of normal (boys, 30 IU/L; girls, 21 IU/L). Results: Of 372 children with DF, 144 (38.7%) had liver-injury. Risk of liver-injury and aminotransferase levels increased with DF severity (p<0.001). Recommended ALT and aspartate aminotransferase (AST) cut-off at ${\geq}1,000IU/L$ had sensitivity 4.8% (5/105), specificity 99.3% (265/267) for detection of severe DF. In children with ALT and AST <1,000 IU/L (n=365), the area under receiver operating curves for prediction for severe DF, were 0.651 (95% confidence interval [CI], 0.588-0.714; p<0.001) for ALT and 0.647 (95% CI, 0.582-0.712; p<0.001) for AST. Serum ALT at 376 IU/L and AST at 635 IU/L had sensitivity and specificity comparable to ${\geq}1,000IU/L$ for defining severe DF. Conclusion: Liver-injury is common in DF. The ALT and AST levels increase with DF severity. ALT and AST levels of ${\geq}1,000IU/L$ could be lowered to 376 IU/L and 635 IU/L respectively for defining severe DF.

• BMB Reports
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• v.34 no.5
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• pp.402-407
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• 2001

Based on the currently proposed toxicity model for the different Bacillus thuringiensis Cry $\delta$-endotoxins, their pore-forming activity involves the insertion of the ${\alpha}4-{\alpha}5$ helical hairpin into the membrane of the target midgut epithelial cell. In this study, a number of polar or charged residues in helix 4 within domain I of the 65-kDa dipteranactive Cry11A toxin, Lys-123, Tyr-125, Asn-128, Ser-130, Gln-135, Arg-136, Gln-139 and Glu-141, were initially substituted with alanine by using PCR-based directed mutagenesis. All mutant toxins were expressed as cytoplasmic inclusions in Escherichia coli upon induction with IPTG. Similar to the wild-type protoxin inclusion, the solubility of each mutant inclusion in the carbonate buffer, pH 9.0, was relatively low When E. coli cells, expressing each of the mutant proteins, were tested for toxicity against Aedes aegypti mosquito-larvae, toxicity was completely abolished for the alanine substitution of arginine at position 136. However, mutations at the other positions still retained a high level of larvicidal activity Interestingly, further analysis of this critical arginine residue by specific mutagenesis showed that conversions of arginine-136 to aspartate, glutamine, or even to the most conserved residue lysine, also abolished the wild-type activity The results of this study revealed an important determinant in toxin function for the positively charged side chain of arginine-136 in helix 4 of the Cry11A toxin.

• The Korean Journal of Pesticide Science
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• v.2 no.1
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• pp.18-23
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• 1998

In order to obtain leading compound for the development of new pesticide through the organic synthesis of natural products, the synthesis of ricinine, an active compound of Ricinus communis, was established and biological activities of synthetic compounds against insects were examined. The synthetic scheme of ricinine was composed of four steps by the spontaneous condensation of the cyanoacetyl chloride. A modified synthetic process was also estabilshed to enhance the synthetic yield by simple cyclization of ethoxymethylene malononitrile. In the bioassay results of synthetic ricinine and intermediates on four insects, the mortality of ricinine on brown planthopper (BPH, Nilaparvata lugens) and pea weevil(PW, Bruchus rufimanus) was 80% and 75% at the concentration of 1,000 ${\mu}g/ml$ respectively. Chloronorricinine and chlororicinic acid having chloride group in molecular structure gave 60% mortality on two-spotted mite (TSSM, Tetranychus urticae) at the concentration of 500 ${\mu}g/ml$. The mortality of compounds on house mosquito (HM, Culex pipens pallens) was meager at 10 ${\mu}g/ml$ level.

• Archives of Plastic Surgery
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• v.37 no.1
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• pp.31-36
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• 2010

Purpose: The primary goal of palatoplasty is to enable normal speech with harmonious growth of face. Some children who had palatoplasty display typical findings of transverse maxillary deficiency requiring orthodontic widening of the maxilla. Levi (2009) described a cleft palate repair coupled with pedicled buccal fat pad flaps to cover bone exposed areas of the hard palate. Hence we report clinical experiences of cleft palate repair using pedicled buccal fat pad flap. Methods: Four Veau class II and a Veau class I cleft palate patients underwent palatoplasty with buccal fat pad flap by single surgeon from April 2009 to August 2009. Two patients received 2-flap palatoplasty and three patients 1-flap palatoplasty, respectively. After the cleft palate repair, sharp mosquito scissors was placed in the superior buccal sulcus just lateral to the maxillary tuberosity and inserted directly through the mucosa resulting in buccal fat pad extrusion. The elevated flap was moved to cover mucoperiosteal defect in hard palatal area. Results: Five patients underwent primary palatoplasty using buccal fat pad flap. Flap harvest and inset took on average 9 minutes per flap. Mucosal epithelization took 18 days on average. No patients had complications related to the buccal fat pad flap. Conclusion: Buccal fat pad pedicled flap has significant potential to function as an added vascularized tissue layer in cleft palate repair and we can expect better growth of maxilla with this method although longer duration of follow-up was unavailable.

• Parasites, Hosts and Diseases
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• v.47 no.4
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• pp.323-335
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• 2009

A successful experience of lymphatic filariasis control in the Republic of Korea is briefly reviewed. Filariasis in the Republic of Korea was exclusively caused by infection with Brugia malayi. Over the past several decades from the 1950s to 2006, many investigators exerted their efforts to detection, treatment, and follow-up of filariasis patients in endemic areas, and to control filariasis. Mass, combined with selective, treatments with diethylcarbamazine to microfilaria positive persons had been made them free from microfilaremia and contributed to significant decrease of the microfilarial density in previously endemic areas. Significant decrease of microfilaria positive cases in an area influenced eventually to the endemicity of filariasis in the relevant locality. Together with remarkable economic growth followed by improvement of environmental and personal hygiene and living standards, the factors stated above have contributed to blocking the transmission cycle of B. malayi and led to disappearance of this mosquito-borne ancient disease in the Republic of Korea.

• Animal cells and systems
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• v.12 no.3
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• pp.165-170
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• 2008

Cuticular hydrocarbons(CHCs) of the epicuticle wax layer are so far used to differentiate the insects in species and subspecies levels. In this study, four species of malaria vectors(genus Anopheles) were collected from various localities in southern Iran. Twenty specimens of each species were randomly selected and one epicuticular extract was prepared of every five specimens. FID-GC profiles of the extracts did not show any qualitative difference. Using significant difference of CHC mass at retention time(RT) 39.6, the two species of An. sacharovi and An. fluviatilis could be distinguished. Similarly, the two species of An. superpictus & An. sacharovi and An. dthali & An. sacharovi were differentiated by their CHC level at RT 28.5. An. sacharovi was distinguished by integratable peaks at RTs 29.7, 30.6, 30.7, 31 and 32.6 while the other three species just indicated trace peaks at the same RTs. Similarly, An. dthali could be known by an integratable peak at RT 26.2 while An. fluviatilis and An. superpictus indicated trace peaks at the same RT. Integratable peaks and traces at RTs 27.4 and 28.5 were respectively used to differentiate An. superpictus from An. fluviatilis. Lastly, CHC trace amount of An. superpictus at RT 39.6 is another indicator to distinguish it from An. fluviatilis with an integratable peak at the same RT. In harmony with other studies worldwide we hereby report that quantitative analysis of CHCs was successfully applied to differentiate the four Anopheles species of southern Iran.

• The Korean Journal of Zoology
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• v.10 no.2
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• pp.18-22
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• 1967

The routine entomological investigations were carried out in the areas of Kaejeon-Myon, OkkuGun, Cholla Pukdo province in 1964 and Sinchang-Myon, Asan Gun, Chungchong Namdo province in 1965, for the better understanding of the behavior of the anpheline mosquitoes especially of the population density , resting , feedign, and breeding habits. The results are as follows : 1. During the investigation , three species of anopheline mosquitoes were recorded ; Anopheles sinensis Wiedmann 1825, A, sineroides Yamada 1935, and A. yatsushiroensis Miyazaki 1951. A . yatsushiroensis Myiyasaki 1951, so far only recorded in Japan, was recorded for the first time in Korea in 1964 ; the site was Kaejong Myon, Okku Gun, Cholla Pukdo province. 2. Anopheles mosquitoes begin to appear from the middle of April and disappear in October. The date of mosquitoes collected by resting place collection in cow shed are three weeks ahead to the night time cow biting collection. 3. Resting places of anopheline mosquitoes are mainly in cow shed and outdoors which provides high humidity and shadow. 4. The population density of a. sinensis sows a peak in late June and early July in cow shed and by cow biting collection respectively, and another small peak in late August and early September. 5, . the biting activity at night is throughout the night from dusk to dawn, sharply decreasing just before dawn. The peak period was different in each months, 2100-2200 hours in June, 2300-2400 in July , 0300-0400 in August , and 2300-2400 in September. 6. The minimum temperature required for the mosquito's biting activity is 15 $^{\circ}C$ and the optimum is between 24-$25^{\circ}C$ : over $25^{\circ}C$ the activity is decreased. 7.A , sinensis appeared to be zoophilic in Sinchang area but 13 times anthrophophilic in Kaejong area than the former. 8. the light attraction of a. sinensis is significantly low in middle June and early August. The parous rate of A. sinensis caught by cow biting collection appeared higher after midnight (2400-0300 hours) with no relation to the peak period of biting activity by month.

• Korean Journal of Veterinary Research
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• v.57 no.1
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• pp.31-36
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• 2017

Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT ($PRNT_{90}$). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and $PRNT_{90}$ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and $PRNT_{90}$ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.

• Microbiology and Biotechnology Letters
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• v.13 no.2
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• pp.157-162
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• 1985

The cell cultures and crude extracts of Bacillus sphaericus 1593 K-5 and its mutant Spo-Dl216 were respectively bioassayed against Culex pipiens var. pollens mosquito larvae. The B. sphaeriucs 1593 K-5 showed toxic activity against the larvae. LC$_{50}$ values (cells/$m\ell$) was 2.6$\times$10$^2$. Also the LC$_{50}$ ($\mu\textrm{g}$ Protein/$m\ell$) of the crude extract was 10.26. However, B. sphaericus Spo-Dl216 didn't show toxic activity against the larvae. The soluble cytoplasmic toxin in broken B. sphaeriucs 1593k-5 cells was partially purified by gel permeation chromatography and ion exchange chromatography. Among the fractions of the gel permeation chromatography only a single fraction was found to be toxic. LC$_{50}$ values ($\mu\textrm{g}$ protein/$m\ell$) of the active fraction was 0.182. The active fraction of the gel permeation was subjected to ion exchange chromatography. Only a single fraction showed toxic activity and its LC$_{50}$ values ($\mu\textrm{g}$ protein/$m\ell$) was 0.02..02.