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Expression of Fusion Protein with Autographa californica Nuclear Polyhedrosis Virus Polyhedrin and Bacillus thuringiensis cryIA(c) Crystal Protein in Insect Cells (곤충세포주에서 Autographa californica 핵다각체병 바이러스의 다각체 단백질과 Bacillus thuringiensis cryIA(c) 내독소 단백질의 융합 단백질 발현)

  • 제연호;진병래;박현우;노종열;장진희;우수동;강석권
    • Korean journal of applied entomology
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    • v.36 no.4
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    • pp.341-350
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    • 1997
  • We have now constructed a novel recombinant baculovirus producing fusion protein with Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin and Bacillus thuringiensis(Bt) cryIA(c) crystal protein. The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The N-terminal of cryIA(c) gene of Bt subsp. kurstaki HD-73 was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front of intact polyhedrin gene or by insertion into the HindIII site in polyhedrin gene. The recombinant baculoviruses were named as BtrusI or BtrusII, respectively. Although single transcript from the fusion protein gene was apparently observed. BtrusI was produced the two proteins, 92 kDa fusion protein and only polyhedrin. In addition, fusion protein produced by BtrusI did not form polyhedra. Interestingly, however, the cells infected with BtrusII did not show a 33 kDa polyhedrin band as a cells infected with BtrusI. Cells infected with BtrusII were only produced fusion protein, but the polyhedra formed by fusion protein was not observed. To determine the insecticidal toxicity of fusion protein, therefore, Sf9 cells infected with BtrusI were inoculated to Bombyx mori larvae. Sf9 cells infected with BtrusI that expressed the fusion protein caused larval mortality although the insecticidal toxicity was low. In conclusion, our results clearly demonstrated that the fusion protein with polyhedrin and Bt cryIA(c) crystal protein have a insecticida toxicity.

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Antibacterial Characteristics of Silver Nano-Particles Attached to Activated Carbon Filter (은나노를 부착한 활성탄 필터의 제균특성)

  • Heo, Ju-Yeong;Nam, Sang-Yeob;Kang, Jeong-Hee;Song, Ji-Hyeon;Kang, Byung-Ha;Han, Sung
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.21 no.10
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    • pp.583-589
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    • 2009
  • Activated carbon has long been used in purification processes for indoor air quality. However, the bioaerosol removal by activated carbon is not often sufficient to be used in an air control devise. In order to overcome these problems, silver nano-particles have been proposed as an antibacterial agent on the surface of activated carbon. Silver or silver ions have been known for antimicrobial activities. In this study, bioaerosol generated by using an Escherichia coli culture was introduced to a lab-scale column packed with activated carbon (AC) and silver nano-particles attached to activated carbon (Ag-AC). E. coli was almost completely removed in the Ag-AC column, whereas bioaerosol penetrated through the AC column. To determine the antibacterial effect of different filter materials in a full-scale air-handling system, another experiment was conducted using a wind tunnel equipped with a heat exchanger and three filter materials including commercial fabric, AC and Ag-AC. It was found that E. coli proliferated on the surface of the heat exchanger after 5 days, which dramatically increased bioaerosol counts in the effluent air stream. The fabric filter could not control the increased bioaerosol and most of the E. coli penetrated the filter. The bacterial removal efficiency was found to be approximately 45% in the AC filter, while the antibacterial efficiency increased to 70% using the Ag-AC filter. Consequently, the Ag-AC filter can be an effective method to control bioaerosol and improve indoor air quality.

Construction of Modified Bacillus thuringiensis cry1Ac Genes for Transgenic Crop Through Multi Site-directed Mutagenesis

  • Xu, Hong Guang;Roh, Jong-Yul;Wang, Yong;Choi, Jae-Young;Shim, Hee-Jin;Liu, Qin;Tao, Xueying;Woo, Soo-Dong;Jin, Byung-Rae;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.19 no.1
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    • pp.199-204
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    • 2009
  • The newly cloned Bacillus thuringiensis cry1-5 gene showed high activity to both Plutella xylostella and Spodoptera exigua, while cry1Ac only showed high activity against P. xylostella but low to S. exigua. Through the alignment of amino acid sequences between Cry1Ac and Cry1-5, we found 12 different residues in domain I (6 residues) and domain II (6 residues). In this study, the modified cry1Ac gene, which is constructed according to a crop-preferring codon usage, was used as a template to construct mutant B. thuringiensis cry1Ac genes based on cry1-5 gene through multi site-directed mutagenesis. Total 63 various mutant cry genes were obtained at 12 positions randomly. Among them, ten mutant cry genes, whose domain I was totally converted and domain II was randomly, were selected to express in baculovirus expression system as a polyhedrin fusion form. The recombinant proteins were 95 kDa in size and were stably activated as 65 kDa by trypsin. The expressed mutant Cry proteins were applied to bioassays against P. xylostella and S. exigua. All mutants showed high insecticidal activity both to P. xylostella and S. exigua similar to cry1-5. These results suggest that these mutant cry genes might be expected of desirable cry genes for introduction to transgenic crops.

The Surgical Treatment in Type III Acromioclavicular Dislocation Patients Over 45 Years - Primary Clavicular Lateral End Resection Method vs. Conventional Acromioclavicular Joint Reduction Method - (45세 이상의 제 3형 견봉쇄골 관절 탈구 환자의 수술적 치료 - 일차적 쇄골 외측단 절제 술식과 고식적인 견봉쇄골 관절 정복 술식의 비교 -)

  • Moon Eun-Sun;Bae Bong-Hyun;Choi Jin;Kim Myung-Sun
    • Clinics in Shoulder and Elbow
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    • v.8 no.2
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    • pp.88-96
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    • 2005
  • Purpose: To compare and analyze the outcome of primary clavicular lateral end resection method and conventional acromioclavicular (AC) joint reduction method in type III AC dislocation patients over 45 years. Materials and Methods: This study was performed on selected 24 cases of type III AC dislocation patients, over 45 years of age, operated at our hospital from 1998 to 2002. Group I consist of 12 patients who underwent primary clavicular lateral end resection methods (average age: 54.3 years$(45{\sim}72)$). Group II consist of 7 patients using Bosworth methods and 5 patients using Phemister methods (average age: 54.4 years$(45{\sim}71)$). Clinical outcome was evaluated by Weaver and Dunn method. Radiological results were compared by measuring coracoclavicular distance between normal and injured side. Results: As clinical outcome, good was 10 cases(83%); fair 2(17%) in Group I, and good 6(50%); fair 3(25%); poor 3(25%) in Group II. In contrast, the difference of coracoclavicular distance was not statistically significant between two groups before or after surgery, and last follow up. At the last follow up, there was no special correlation between the difference of coracoclavicular distance and clinical outcome. Conclusion: We considered that primary clavicular lateral end resection may be effective for prevention of arthrosis in AC joint in type III AC dislocation patients over 45 years.

Electrical Characteristics of ZnO Surge Arrester Elements Subjected to the Mixed DC and 60[Hz] AC Voltages (직류+60[Hz] 교류 중첩전압에 대한 ZnO 피뢰기 소자의 전기적 특성)

  • Lee, Bok-Hee;Yang, Soon-Man
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.26 no.4
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    • pp.41-47
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    • 2012
  • This paper deals with the electrical characteristics related to power loss, equivalent resistance, and leakage currents flowing through new and deteriorated zinc oxide(ZnO) arrester elements subjected to the mixed DC and 60[Hz] AC voltages. The test specimens were deteriorated by 8/20[${\mu}s$] impulse current of 2.5[kA]. The leakage current-applied voltage($I-V$) characteristic curves of ZnO surge arrester elements were measured as a parameter of the ratio of the peak of 60[Hz] AC voltage to the peak of total voltage. As a consequence of test results, in case of the same applied voltage, the leakage currents flowing through the deteriorated ZnO arrester elements were higher than those flowing through the new ZnO surge arrester elements. The cross-over phenomenon in $I-V$ curves of ZnO surge arrester elements measured as a parameter of the mixed ratio of DC and AC voltages was observed at the low current domain. The effect of DC voltage on the leakage current flowing through ZnO surge arrester elements is pronounced at the same magnitude of test voltages. In addition, the larger the applied number of 8/20[${\mu}s$] impulse current of 2.5[kA] is, the greater the power loss is, in particular, the more severe the power loss increases at higher applied voltages.

Compound K, a Metabolite of Ginsenoside Rb1, Inhibits Passive Cutaneous Anaphylaxis Reaction in Mice

  • Bae, Eun-Ah;Trinh, Hien Trung;Yoon, Hae-Kyung;Kim, Dong-Hyun
    • Journal of Ginseng Research
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    • v.33 no.2
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    • pp.93-98
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    • 2009
  • To understand the anti-allergic mechanism of compound K, which is a metabolite of ginsenoside Rb1, a main constituent of the root of Panax ginseng C.A. Meyer (family Araliaceae), its inhibitory effect against IgE-antigen complex IAC)-induced passive cutaneous anaphylaxis (PCA) reaction in mice and mRNA and protein expressions of allergic cytokines in lAC-stimulated RBL-2H3 cells were investigated. Orally administered ginsenoside Rb1 more potently inhibited PCA reaction when administered at 5 h prior to the lAC treatment than when administered at I h before. However, compound K orally administered 1 h before lAC treatment showed a more potent anti-PCA reaction effect than when treated at 5 h before. Orally administered ginsenoside Rb1 more potently inhibited PCA reaction induced by lAC in mice than intraperitoneally treated one, apart from orally administered its metabolite, compound K, which was more potent than the orally administered one. The compound K, a metabolite of ginsenoside Rb1, inhibited mRNA and protein expressions of IL-4 and TNF-${\alpha}$ and the activation of their transcription factor NF-$\kappa$B and MAPK in lAC-stimulated RBL-2H3 cells. These findings suggest that orally administered ginsenoside Rb1 may be dependent on its metabolism by intestinal microflora in the intestine and the compound K may improve allergic diseases by the inhibition of IL-4 and TNF-${\alpha}$ expresseion.

Comparison of Anti-inflammatory effects between Artemisia capillaris and Artemisia iwayomogi by extraction solvents (인진호(茵蔯蒿)와 한인진(韓茵蔯)의 추출용매별 항염증 효능 비교)

  • Noh, Dongjin;Choi, Jin Gyu;Hong, Soon-Sun;Oh, Myung Sook
    • The Korea Journal of Herbology
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    • v.33 no.3
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    • pp.55-61
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    • 2018
  • Objectives : Artemisia capillaris Thunberg (AC) and Artemisia iwayomogi Kitamura (AI) have been used without distinguishment since ancient times due to similar appearance. In this study, we compared the inhibitory effects of AC and AI on the expression of inflammatory cytokines induced by lipopolysaccharide (LPS) in murine macrophages. Methods : AC and AI were extracted by reflux with distilled water (DW) and 70% ethanol (EtOH). We investigated the inhibitory effects of AC and AI on the expression of nitric oxide (NO), inducible NO synthase (iNOS) and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) induced by LPS in macrophages. Results : Firstly, yield of the samples was higher in order of Artemisia iwayomogi DW Extract (AID), Artemisia iwayomogi 70% EtOH Extract (AIE), Artemisia capillaris DW Extract (ACD) and Artemisia capillaris 70% EtOH Extract (ACE). All of the samples were not toxic in macrophages. The inhibitory effect of the samples on LPS-induced NO expression was stronger in the order of AIE, ACE, AID and ACD. The inhibitory effect of the samples on LPS-induced inducible iNOS expression was stronger in the order of AIE, ACE and AID. Effect of ACD was same with that of AID. In addition, inhibitory effect of the samples on LPS induced $TNF-{\alpha}$expression wes stronger in the order of AIE, ACE, AID and ACD. Conclusion: These results showed that AI would be more effective than AC and 70% EtOH would be more effective than DW as an extraction solvent in inflammatory diseases.

Anticolitic Effect of the Rhizome Mixture of Anemarrhena asphodeloides and Coptidis chinensis (AC-mix) in Mice

  • Jang, Se-Eun;Jeong, Jin-Ju;Hyam, Supriya R.;Han, Myung Joo;Kim, Dong-Hyun
    • Biomolecules & Therapeutics
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    • v.21 no.5
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    • pp.398-404
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    • 2013
  • During a screening program to search the anticolitic herbal medicines, 80% ethanol extract of the rhizome of Anemarrhena asphodeloides (AA) was found to potently inhibit the expression of proinflammatory cytokines TNF-${\alpha}$ and IL-1${\beta}$, as well as the activation of NF-${\kappa}B$ in LPS-stimulated colonic macrophages, followed by that of the rhizome of C. chinensis (CC). AA also potently inhibited TNBS-induced colitic markers, shortening of the colon and increase of macroscopic score, myeloperoxidase activity, TNF-${\alpha}$, IL-1${\beta}$, and IL-6, in mice. The synergistic effect of CC against the anticolitic effect of AA was investigated. CC synergistically inhibited the anticolitic effect of AA. AC-mix (AA+CC, 1:1) potently inhibited them. AC-mix also inhibited the activation of NF-${\kappa}B$, as well as the expression of TNF-${\alpha}$, IL-1${\beta}$, IL-6, iNOS and COX-2. The effects of AC-mix against oxazolone-induced colitis were investigated in mice. AC-mix also potently inhibited oxazolone-induced inflammatory markers, colon shortening, macroscopic score, myeloperoxidase activity, NF-${\kappa}B$ activation and proinflammatory cytokines. Overall, the anti-colitic effect of AC-mix was superior to that of mesalazine. Based on these findings, AC-mix may improve colitis by inhibiting NF-${\kappa}B$ activation.