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Functional Expression of Candida antarctica Lipase A in Pichia a pastoris and Escherichia coli (Pichia pastoris와 Escherichia coli를 이용한 Candida antarctica Lipase A의 기능적 발현)

  • Park, Hye-Jung;Kim, Yong-Hwan
    • KSBB Journal
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    • v.24 no.4
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    • pp.341-346
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    • 2009
  • Candida Antarctica lipase A (CalA) has been used because of its suitability in industrial applications. CalA has unique features capable to accept tertiary and sterically hindered alcohols among many hydrolases. CalA gene was cloned and constructed in expression vector such as pColdIII/CalA and $pPICZ{\alpha}A$/CalA. The gene encoding pColdIII/CalA was functionally expressed in the cytoplasm of Escherichia coli $Origami^{TM}$ B (DE3) cells. The plasmid $pPICZ{\alpha}A$/CalA linearized by BstX I was integrated into 5'AOX1 region of the chromosomal DNA and was functionally expressed in the methyl atrophic yeast Pichia pastoris. Expressed CalA in P. pastoris (0.7 Unit/mL) showed 35 times higher activity than that in E. coli expression system (0.02 Unit/mL).

True Posterior Communicating Artery Aneurysms with High Risk of Rupture despite Very Small Diameter

  • Shin, Dong Gyu;Park, Jaechan;Kim, Myungsoo;Kim, Byoung-Joon;Shin, Im Hee
    • Journal of Korean Neurosurgical Society
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    • v.65 no.2
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    • pp.215-223
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    • 2022
  • Objective : This retrospective study investigated the clinical and angiographic characteristics of ruptured true posterior communicating artery (PCoA) aneurysms in comparison with junctional PCoA aneurysms presenting with a subarachnoid hemorrhage. Methods : The medical records and radiological data of 93 consecutive patients who underwent three-dimensional rotational angiography and surgical or endovascular treatment for a ruptured junctional or true PCoA aneurysm over an 8-year period were examined. Results : The maximum diameter of the ruptured true PCoA aneurysm (n=13, 14.0%) was significantly smaller than that of the ruptured junctional PCoA aneurysms (n=80, 4.45±1.44 vs. 7.68±3.36 mm, p=0.001). In particular, the incidence of very small aneurysms <4 mm was 46.2% (six of 13 patients) in the ruptured true PCoA aneurysm group, yet only 2.5% (two of 80 patients) in the ruptured junctional PCoA aneurysm group. Meanwhile, the diameter of the PCoA was significantly larger in the true PCoA aneurysm group than that in the junctional PCoA aneurysm group (1.90±0.57 vs. 1.15±0.49 mm, p<0.001). In addition, the ipsilateral PCoA/P1 ratio was significantly larger in the true PCoA aneurysm group than that in the group of a junctional PCoA aneurysm (mean PCoA/P1 ratio±standard deviation, 2.67±1.22 vs. 1.14±0.88; p<0.001). No between-group difference was identified for the modified Fisher grade, clinical grade at admission, and 3-month modified Rankin Scale score. Conclusion : A true PCoA aneurysm was found to be associated with a larger PCoA and ruptured at a smaller diameter than a junctional PCoA aneurysm. In particular, the incidence of a ruptured aneurysm with a very small diameter <4 mm was significantly higher among the patients with a true PCoA aneurysm.

Differential Response of Surfactant Protein-A Genetic Variants to Dexamethasone Treatment (덱사메타손 처치에 따른 폐 표면 활성 단백질-A 유전자 변이의 반응의 차이점에 관한 연구)

  • Kim, Eul Soon;Lee, In Kyu;Oh, Myung Ho;Bae, Chong Woo
    • Clinical and Experimental Pediatrics
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    • v.46 no.4
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    • pp.335-339
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    • 2003
  • Purpose : Surfactant protein A(SP-A) is involved in surfactant physiology and structure, and plays a major role in innate host defense and inflammatory processes in the lung. Steroid therapy is widely used for mothers who threaten to deliver prematurely and also used commonly in the management of preterm infants with chronic lung disease. Two SP-A genes(SP-A1, SP-A2) and several alleles have been characterized for each SP-A gene in human. Preliminary evidence indicates that differences may exist among alleles in response to Dexamethasone(Dexa) and that the SP-A 3'UTR plays a role in this process. We studied whether 3'UTR-mediated differences exist among the most frequently found SP-A alleles in response to Dexa. Methods : Constructs containing the 3'UTR from eight different SP-A alleles were made using luciferase as a the reporter gene. These constructs were driven by the SV40 promotor and were transfected along with a transfection control vector in H441 cells that express SP-A. The activity of the reporter gene in the presence or absence of Dexa(100 nM) treatment was measured. All the experiments for the eight SP-A alleles studied, were performed in triplicate and repeated five times. The results were normalized to the transfection control. Results : Expression of alleles of 6A3, 6A, 1A were significantly decreased in response to Dexa. Conclusion : Three UTR mediated differences exist among human SP-A variants both in the basal expression and in response to Dexa. These genotype-dependent differences may point to a need for a careful consideration of individual use of steroid treatment in the prematurely born infant.

A NEW PROOF OF MACK'S CHARACTERIZATION OF PCS-ALGEBRAS

  • Kim, Hyoung-Soon;Woo, Seong-Choul
    • Communications of the Korean Mathematical Society
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    • v.18 no.1
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    • pp.59-63
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    • 2003
  • Let A be a $C^*$-algebra and $K_{A}$ its Pedersen's ideal. A is called a PCS-algebra if the multiplier $\Gamma(K_{A})\;of\;K_{A}$ is the multiplier M(A) of A. J. Mack [5]characterized PCS-algebras by weak compactness on the spectrum of A. We give a new simple proof of this Mack's result using the concept of semicontinuity and N. C. Phillips' description of $\Gamma(K_{A})$.

한국인으로부터 분리한 비피더스균의 특성과 Bifidobacterium longum A-2의 임상실험에 관한 연구

  • Kim, Yeong-Chan
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 1998.10a
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    • pp.83-106
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    • 1998
  • This study was conducted to investigate the probiotics(acid and bile resistance, fermentation properties, viability, cholesterol assimilation, antimicrobial activity, antimutagenicity, and immunoactivation) of the strains of bifidobacteria isolated from healthy Koreans and to investigate the effects of oral administration of Bifidobacterium longum A-2 on the fecal microflora, ${\beta}-glucuronidase$ activity, pH values, Ammonia concentration. The experimental results are summarized as follows: The probiotics were tested for 23 strains including three commer챠al strains as controls. Compared to other strains, strains of A-2 and A-9 showed more acid resistance whereas A-2, A-5, A-13, A-14, A-18 and A-22 showed excellent bile resistances. The properties of bifidobacteria during fermentation were tested. Strains of A-1, A-2, A-3, A-4, A-6, and A-23 resulted in less than pH 4.5 and titratable acidity over 0.90 after 24 hr of fermentation. When the strains of A-2 was grown with glucose, maltose, and fructooligosaccharide, the acetic acid production were higher than with sorbitol and mannitol. The storage stability of the strains of A-2 and A-22 were tesed, indicating the strain A-2 was more stable over 10 days of storage at both $4^{\circ}C$ and $20^{\circ}C$ than A-22. The strains of A-8, A-10, A-11, A-12 and A-20 assimilated more than 30% of cholesterol included in the media. The strains of A-1 and A-2 showed antimicrobial activity against Sta. aureus. The antimutagenicity of the strains were also tested, showing that the mutation was suppressed more by three strains(A-2, A-12, and A-23). In addition, strain A-5 improved immunological activity(phagocytosis, $TNF-{\alpha}$, IL-6) more than other strains. In the effects of oral administration of Bif. longum A-2, the number of fecal bifidobacteria was siginificantly increased(p<0.01) and the level of fecal ${\beta}-glucuronidase$ also was siginificantly reduced(p<0.05). However there were no siginificant differences in the level of Lαctobacilli, Enterobacteriaceae, Clostridium perfringens, pH and ammonia by the administration. The results suggested that Bif. longum A-2 may be met the criteria for probiotics culture.

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Antineoplastic Effect of Several Herbal Medicine Mixtures on SNU-80 Anaplastic Thyroid Carcinoma Cell Line (수종 한약 복합물의 역형성갑상선암세포 SNU-80에 대한 항암효과)

  • Yeo, Hyun-Soo;Lee, Min-Hye;Choi, You-Kyung;Jun, Chan-Young;Park, Jong-Hyeong
    • The Journal of Internal Korean Medicine
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    • v.35 no.4
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    • pp.416-427
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    • 2014
  • Objectives: The purpose of this study was to investigate the antineoplastic effect of several herbal medicine mixtures (compositions of Astragalus membranaceu, Angelica gigas, Trichosanthes kirilowii, Panax ginseng, Rhus verniciflua Stokes) on the SNU-80 anaplastic thyroid carcinoma cell line. Methods: MTT assay was used to examine whether our herbal medicine mixtures decreased cell growth rate of SNU-80. Wound healing assay and Transwell invasion assay was performed to investigate whether our herbal medicine mixtures affect the migration and invasion of anaplastic cancer cells, SNU-80. ELISA assay was performed to know if our herbal medicine mixtures suppressed the expression of pro-invasive molecules, such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) secreted from SNU-80. Results: MTT assay demonstrated that A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas :T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 strongly suppressed the growth of SNU-80. Wound healing assay demonstrated that A. membranaceus:A. gigas=3:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the migration of SNU-80. Transwell invasion assay demonstrated that A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii =1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng :R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the invasion of SNU-80. ELISA assay demonstrated that A. membranaceus :A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 suppressed the expression of VEGF. Also, A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus :A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes =1:1:1:1:1 or 3:1:1:1:1 suppressed the expression of MMP-2. Conclusions: The results obtained in this study suggest that several herbal medicine mixtures suppresse the growth and inhibit the migration and invasion of SNU-80, which is anaplastic thyroid cancer cells. Especially, A. membranaceus:A. gigas: T. kirilowii=1:1:1 mixture had a stronger anti-cancer effect.

Cloning and functional expression of a cecropin-A gene from the Japanese oak silkworm, Antheraea yamamai (천잠 cecropin-A 유전자 클로닝 및 재조합 발현)

  • Kim, Seong-Ryul;Choi, Kwang-Ho;Kim, Sung-Wan;Goo, Tae-Won;Hwang, Jae-Sam
    • Journal of Sericultural and Entomological Science
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    • v.52 no.1
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    • pp.45-51
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    • 2014
  • A cecropin-A gene was isolated from the immunized larvae of the Japanese oak silkworm, Antheraea yamamai and designed Ay-CecA. The complete Ay-CecA cDNA consists of 419 nucleotides with 195 bp open reading frame encoding a 64 amino acid precursor that contains a putative 22-residue signal peptide, a 4-residue propetide and a 37-residue mature peptide with a theoretical mass of 4046.81. The deduced amino acid sequence of the peptide evidenced a significant degree of identity (62 ~ 78% identity) with other lepidopteran cecropins. Like many insect cecropin, Ay-CecA also harbored a glycine residue for C-terminal amidation at the C-end, which suggests potential amidation. To understand this peptide better, we successfully expressed bioactive recombinant Ay-CecA in Escherichia coli that are highly sensitive to the mature peptide. For this, we fused mature Ay-CecA gene with insoluble protein ketosteroid isomerase (KSI) gene to avoid the cell death during induction. The fusion KSI-CecA protein was expressed as inclusion body. The expressed fusion protein was purified by Ni-NTA immobilized metal affinity chromatography (IMAC), and cleaved by cyanogen bromide (CNBr) to release recombinant Ay-CecA. The purified recombinant Ay-CecA showed considerably antibacterial activity against Gram-negative bacteria, E. cori ML 35, Klebsiella pneumonia and Pseudomonas aeruginosa. Our results proved that this peptide with a potent antibacterial activity may play a role in the immune response of Japanese oak silkworm.

Genetic Control of Asexual Sporulation in Fusarium graminearum

  • Son, Hokyoung;Kim, Myung-Gu;Chae, Suhn-Kee;Lee, Yin-Won
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.15-15
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    • 2014
  • Fusarium graminearum (teleomorph Gibberella zeae) is an important plant pathogen that causes head blight of major cereal crops such as wheat, barley, and rice, as well as causing ear and stalk rot on maize worldwide. Plant diseases caused by this fungus lead to severe yield losses and accumulation of harmful mycotoxins in infected cereals [1]. Fungi utilize spore production as a mean to rapidly avoid unfavorable environmental conditions and to amplify their population. Spores are produced sexually and asexually and their production is precisely controlled. Upstream developmental activators consist of fluffy genes have been known to orchestrate early induction of condiogenesis in a model filamentous fungus Aspergillus nidulans. To understand the molecular mechanisms underlying conidiogenesis in F. graminearum, we characterized functions of the F. graminearum fluffy gene homologs [2]. We found that FlbD is conserved regulatory function for conidiogenesis in both A. nidulans and F. graminearum among five fluffy gene homologs. flbD deletion abolished conidia and perithecia production, suggesting that FlbD have global roles in hyphal differentiation processes in F. graminearum. We further identified and functionally characterized the ortholog of AbaA, which is involved in differentiation from vegetative hyphae to conidia and known to be absent in F. graminearum [3]. Deletion of abaA did not affect vegetative growth, sexual development, or virulence, but conidium production was completely abolished and thin hyphae grew from abnormally shaped phialides in abaA deletion mutants. Overexpression of abaA resulted in pleiotropic defects such as impaired sexual and asexual development, retarded conidium germination, and reduced trichothecene production. AbaA localized to the nuclei of phialides and terminal cells of mature conidia. Successful interspecies complementation using A. nidulans AbaA and the conserved AbaA-WetA pathway demonstrated that the molecular mechanisms responsible for AbaA activity are conserved in F. graminearum as they are in A. nidulans. F. graminearum ortholog of Aspergillus nidulans wetA has been shown to be involved in conidiogenesis and conidium maturation [4]. Deletion of F. graminearum wetA did not alter mycelial growth, sexual development, or virulence, but the wetA deletion mutants produced longer conidia with fewer septa, and the conidia were sensitive to acute stresses, such as oxidative stress and heat stress. Furthermore, the survival rate of aged conidia from the F. graminearum wetA deletion mutants was reduced. The wetA deletion resulted in vigorous generation of single-celled conidia through autophagy-dependent microcycle conidiation, indicating that WetA functions to maintain conidia dormancy by suppressing microcycle conidiation in F. graminearum. In A. nidulans, FlbB physically interacts with FlbD and FlbE, and the resulting FlbB/FlbE and FlbB/FlbD complexes induce the expression of flbD and brlA, respectively. BrlA is an activator of the AbaA-WetA pathway. AbaA and WetA are required for phialide formation and conidia maturation, respectively [5]. In F. graminearum, the AbaA-WetA pathway is similar to that of A. nidulans, except a brlA ortholog does not exist. Amongst the fluffy genes, only fgflbD has a conserved role for regulation of the AbaA-WetA pathway.

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The effect of visual factors on the reader's preference in the book cover design -Focused on the literary books (책표지 디자인에서 시각적 요소가 독자 선호도에 미치는 영향 -문학도서를 중심으로)

  • 남미현;백진경
    • Archives of design research
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    • v.17 no.1
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    • pp.329-338
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    • 2004
  • This study did derive a characteristic of a cover design through considering a concept, a function and components of a cover design based on a literature study and through analyzing cases of the best-seller cover designs for the literature section from 1999 to 2002. In terms of a reader's preference for a cover design according to an age and sex of a reader, it tried to suggest a condition and a method of a book-cover design through a questionnaire survey and the image evaluation. In a survey of the preference for a cover design, there was the difference of interest degree in a kind of a book, a place to buy a book and a cover design according to an age and sex. In addition, it was shown that a woman prefers the stylish typography, and both a man and a woman prefer the cool color. However, except these two, the reason why there is no difference in preference for visual elements are that a cover design is an integrative image comprising several elements. Accordingly, through the Semantic Differential Method, it carried out the evaluation as to a cover design as an integrative image. As a result of that, an abstract image was shown to be difficult to convey a meaning because of its fall in clarity. Further, a cover design mainly comprising the stylish typography gained the high evaluation in unique and clarity. It is thought to be attributable to the easiness to achieve a goal of seeing and reading in a cover design in which an image and the typography are balanced, rather than the excessive visual. Even though the visual is excellent, there was difference from what having been excellent in conveying a meaning, which is a functional aspect. This study tried to examine, limiting it only to a visual element. It will be hereafter required to proceed with a study on the influence on a reader's preference according to material, bookbinding and a form of book which are elements comprising a cover design.

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A Study on Proving RMF A&A in Real World for Weapon System Development (무기체계 개발을 위한 RMF A&A의 실증에 관한 연구)

  • Cho, Kwangsoo;Kim, Seungjoo
    • Journal of the Korea Institute of Information Security & Cryptology
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    • v.31 no.4
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    • pp.817-839
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    • 2021
  • To manage software safely, the military acquires and manages products in accordance with the RMF A&A. RMF A&A is standard for acquiring IT products used in the military. And it covers the requirements, acquisition through evaluation and maintenance of products. According to the RMF A&A, product development activities should reflect the risks of the military. In other words, developers have mitigated the risks through security by design and supply chain security. And they submit evidence proving that they have properly comply with RMF A&A's security requirements, and the military will evaluate the evidence to determine whether to acquire IT product. Previously, case study of RMF A&A have been already conducted. But it is difficult to apply in real-world, because it only address part of RMF A&A and detailed information is confidential. In this paper, we propose the evidence fulfilling method that can satisfy the requirements of the RMF A&A. Furthermore, we apply the proposed method to real-world drone system for verifying our method meets the RMF A&A.