• Title/Summary/Keyword: Phase contrast microscope

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Characterization of New Bacillus thuringiensis Isolated with Bioactivities to Tobacco Cutworm, Spodoptera litura (Lepidoptera: Noctuidae) (담배거세미나방에 살충효과를 나타내는 새로운 Bacillus thuringiensis 균주의 특성)

  • Kim, Da-A;Kim, Jin-Su;Kil, Mi-Ra;Paek, Seung-Kyoung;Choi, Su-Yeon;Jin, Da-Yong;Youn, Young-Nam;Hwang, In-Cheon;Yu, Yong-Man
    • Korean journal of applied entomology
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    • v.47 no.1
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    • pp.87-93
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    • 2008
  • Bacillus thuringiensis with selected high toxicities against tobacco cutworm, Spodoptera litura were isolated from domestic soils. When being observed under a phase-contrast microscope, the insecticidal crystal proteins were showed a bipyramidal crystal types. New CAB 109 isolate was identified to B. thuringiensis subsp. aizawai in the H serotype. As a results of insecticidal activities between CAB 109 isolate and 3 existing ready-made products against 3rd larva of S. litura, CAB 109 isolate showed 100% mortality with spore concentration $(1.3{\times}10^7cfu/ml)$. It was a very high insecticidal activity compared with a existing ready-made B. t. products. $LD_{50}$ values of CAB 109 isolate was $9.78{\times}10^5,\;6.87{\times}10^6\;and\;1.83{\times}10^7cfu/ml$ spore concentration against 2nd, 3rd and 4th larva of S. litura, respectively. Unlike Plutella xylostella, S. litura was slowly died after application up to 7 days. The weight of S. litura larva applied with CAB 109 isolate were 6-7 times less than controlled group. Even though it didn't die, it did not grow into next larva. The result observed with scanning electron microscope was that CAB 109 isolate of B. t. aizawai formed a typical bipyramidal crystal protein type. Otherwise, when CAB 109 isolate was examined with SDS-PAGE and with trypsin, there was no difference between CAB 109 strain and ready-made products of B. thuringiensis.

A Study on the Chitin and Protein Contents in Shells of 5 Marine Crustaceans (5종(種)의 해산(海産) 갑각류각피(甲殼類殼皮)에서의 Chitin 및 단백질함량(蛋白質含量)에 관한 연구(硏究))

  • Lee, Mee-Sook;Seo, Jung-Sook;Mo, Su-Mi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.13 no.3
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    • pp.307-312
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    • 1984
  • The dried pure shells deprived of soft tissues were subjected to analysis of chitin-protein complexes from 5 species of marine crustaceans, including 2 species of crabs and 3 species of shrimps. The protein fractions were obtained from chitin-protein complexes under the varying conditions of extractions and the crude chitin was prepared from the shells by the sulfurous acid process. The crude chitin was purified through the extraction with several organic solvents such as dimethyl-acetamide, N-methylpyrrolidone. The purified chitin was also examined using the phase contrast microscope. Total protein contents of the shells were diverse, showing 9.6% for Portunus trituberculatus, 3.1%, Charybdis bimaculata, 9.4%, Penaeus japonicus, 10.9%, Metapenaeus intermedius and 5.8%, Squilla oratoria. Covalently bound protein varied with species from 2.1% for Charybdis bimaculata to 9.9% for Metapenaeus intermedius. The puified chitin contents of the shells were shown to 21.1% for Portunus tritube rculatus, 6.2%, Charybdis bimaculata, 20.2%, Penaeus japonicus, 27.1%, Metapenaeus intermedius and 25.5%, Squilla oratoria. Exceptionally low analytical value obtained with Charybdis bimaculata are supposed to be due to the very young subjects. The ratios of chitin to covalently bound protein in the shells were various such as 2.7 to 1 for Portunus trituberculatus, Penaeus japonicus and Metapenaeus intermedius, 3.1 to 1, Charybdis bimaculata and 6.1 to 1, Squilla oratoria. The microscope finding of the purified chitin showed the filamentous form in all the specimen.

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The effect of safflower seed fraction extract on periodontal ligament fibroblast and MC3T3-E1 cell in vitro (홍화씨 분획 추출물이 치주인대 섬유아세포와 MC3T3-E1 세포에 미치는 영향)

  • Huh, Ji-Sun;Kang, Jung-Hwa;Yoo, Yun-Jung;Kim, Chang-Sung;Cho, Kyoo-Sung;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.31 no.4
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    • pp.833-846
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    • 2001
  • Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.

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Characterization of Biopesticides (Bacillus thuringiensis) Produced in Korea (국내에서 생산된 Bacillus thuringiensis 살충제의 특성)

  • Kil, Mi-Ra;Kim, Da-A;Choi, Su-Yeon;Paek, Seung-Kyoung;Kim, Jin-Su;Jin, Da-Yong;Hwang, In-Chon;Yu, Yong-Man
    • The Korean Journal of Pesticide Science
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    • v.11 no.3
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    • pp.201-209
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    • 2007
  • Characteristics of the 5 biopesticides that included Bacillus thuringiensis and on the domestic markets were investigated. These products were contained different strains of B. thuringiensis, for examples; product A and E was B. thuringiensis subsp aizawai; product B was B. thuringiensis; product C was B. thuringiensis Berline var. kurstaki; product D was B. thuringiensis var. kurstaki. Number of active spores were counted because they could influence the bio-activity against target pests. Only product C are contained the fixed quantity as its label, however, product D and E were a tenth part, and product A and B were a hundredth part of their descriptions. The pHs of product A and B were measured 3.67 and 3.73, and C, D and E were 5, respectively. Typical bypyramidal crystals produced from B. thuringiensis was found in only product C under a phase contrast microscope. For the uniform formulation of products that conformed whether B. thuringiensis were equally spreaded on the crops, B. thuringiensis in the C, D and E were equally grown on the nutrient agar medium As a results, product A were more different from product C than any other products. When product A and C were bioassayed against different larval stages of diamondback moth, their mortalities with spraying application were showed 100% after 48 hours.

THE EFFECTS OF VARIOUS TETRACYCLINE HCL CONCENTRATION TREATED ROOW SURFACES ON PROLIFERATION AND SPREADING OF PERIODONTAL LIGAMENT CELLS (다양한 농도의 테트라사이클린로 처리된 치근면이 치주인대세포의 증식과 전개에 미치는 영향)

  • Jung, Oh-Chul;Sun, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.24 no.3
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    • pp.581-596
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    • 1994
  • This in vitro study was undertaken to obtain optimal tetracycline concentration that aids proliferation and spreading of human periodontal ligament cells, for clinical application in root surfaces of periodontally diseased teeth. Periodontal ligament cells used in this study were obtained from explants of periodontal ligament of 1st premolar teeth which were extracted for the purpose of orthodontic treatment. The cells were cultured in Dulbecco's Modified Eagle Medium(DMEM) supplemented with 100 U/ml penicillin, $100\;{\mu}g/ml$ streptomycin and 10% FBS at $37^{\circ}C$, 100% humidity, 5% $CO_2-95%$ air. Cells were used between the third to 4th passage. After root planing of periodontally extracted teeth, the root slabs were cut with carborundum disk. In the cell proliferation experiment, experimental groups were root planing only group, immersed groups in 25, 50, 75, 100, 150mg/ml aqueous solution of Tetracycline HCl followed by a vigorous rinse in PBS. Human PDL cells at concentration of $1{\times}10^5\;cells/ml$ were seeded in each culture well which contained root slabs and incubated for 6 hours. Then, all of the root slabs were moved into new 24 culture well and incubated 24, 48 and 72 hours. The cell counting was done by inverted phase contrast microscope after trypsinization. The following results were obtained. The cell number was increased in order root planing only group, 25, 150, 50, 75, 100mg/ml of Tetracycline HCl treated group in 24, 48 and 72 hours. The maximal cell number was obtained when the root slabs were immersed in solution with 100mg/ml of Tetracycline HCl. There were statistically significant between the root planing only group and 75, 100 mg/ml of Tetracycline HCl treated group in 24 hours, between the root planing only group and 100mg/ml of Tetracycline HCl treated group in 48 hours, between the root planing only group and 50, 75, 100mg/ml of Tetracycline HCl treated group, between 25 and 100mg/ml of Tetracycline HCl treated group in 72 hours(p<0.05). In the cell spreading experiment, after 30 minutes of incubated, in the root planing only group, the cells were generally round in shape. The cell surface was mostly covered with blebs. The cells started to attach to root surface by cytoplasmic extension in 50, 100mg/ml of Tetracycline HCl treated groups, more numerous cells attached to root surface than root planing only group. Many orifices of dentinal tubule were exposed, cells showed radially spreaded cytoplasm and unspreaded central region of the cell was covered with blebs. After 6 hours of incubation, in the root planing only group, cells showed radially spreaded cytoplasm and were attached flat appearance. In 50, 100mg/ml of Tetracycline HCl treated groups, cellular margin was concaved and cytoplasm showed elongated appearance with polarity. After 24 hours of incubation, in the root planing group, cells showed characteristic polarity. In 50, 100mg/ml of Tetracycline HCl treated groups, cells showed more elongated and spindle - like appearance.

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