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Development of Bursaphelenchus xylophilus-specific antibody for detecting the pine wilt disease
Dae-Weon Lee,A Young Kim,Won Tae Kim,Kook-Jin Lim,Yeon Ho Je,Si-Hyeock Lee,Young Ho Koh 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.05
The pinewood nematode (PWN, Bursaphelenchus xylophilus) causes the pine wilt disease, transmitted to pinewoods by the pine sawyer beetle, Monochamus alternatus. It is very difficult to discriminate B. xylophilus from B. mucronatus. Therefore, it has been necessary to detect PWN-infected trees for the prevention of pine wilt disease transmission in a short time. The development of biomarkers such as DNA and protein is important for diagnosis of B. xylophilus. However, there have been no reports regarding biomarker identifications of B. xylophilus. In this study, polyclonal antisera were raised against whole proteins of B. xylophilus in BalbC mice and were primarily screened with ELISA. Twenty five among over 500 cell lines releasing polyclonal antisera revealed B. xylophilus-specific immuno-reactivity, which was at least three times higher than that of B. mucronatus. Three cell lines among them were secreting monoclonal antibody through further screening. These cell lines only detect about a 33-kDa protein in B. xylophilus in the western blot. These results suggest that these monoclonal antibodies will be useful for the development of diagnostic kit for the pine wilt disease.
Identification of a Novel Pectate Lyase3 from the pine wood nematode, Bursaphelenchus xylophilus
Dae-Weon Lee,Jae Soon Kang,A Young Kim,Si-Hyeock Lee,Young Ho Koh 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10
The pinewood nematode (PWN, Bursaphelenchus xylophilus) is known to be a major pathogen of the pine wilt disease (PWD). However molecular pathology of B. xylophilus is not completely understood, the pathogenecity of PWD is related to cell wall-degrading enzymes such as endoglucanases, expansins and pectate lyases (PELs). Recently, we developed stage-specific expressed tag library of B. xylophilus and identified a novel PEL, Bx-PEL3. We cloned Bx-PEL3 gene with RT-PCR, which showed high similarity to previously reported Bx-PELs. Phylogenetic analysis revealed that PEL3 was much closer to PELs of B. xylophilus than any other PELs. PEL3 has a conserved intron site as found in Bx-PEL2 in the genomic DNA analysis. Quantitative real-time PCR analysis revealed that Bx-PEL1 and Bx-PEL2 were more predominantly expressed than the Bx-PEL3 in B. xylophilus. The difference of expression level among Bx-PELs according to growth condition suggests that each Bx-PEL plays different biochemical role in the pathogenesis of the PWD.
Dae-Weon Lee,Jong Bok Seo,A Young Kim,Won Tae Kim,Si Hyeock Lee,Young Ho Koh 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05
tasks that require nematode extraction and microscopic examination. To develop a more efficient detection method for Bursaphelenchus xylophilus, we first generated monoclonal antibodies (MAbs) specific to B. xylophilus. Among 2,304 hybridoma fusions screened, a hybridoma clone named 3-2A7-2H5 recognized a single protein from B. xylophilus specifically. We finally selected the MAb clone 3-2A7-2H5-D9-F10 (D9-F10) for further studies. To identify the antigenic target of MAb-D9-F10, we analyzed proteins in spots, fractions or bands via nano liquid chromatography electrospray ionization quadrupole ion trap mass spectrometry (nano-LC-ESI-Q-IT-MS). Peptides of galactose-binding lectin-1 of B. xylophilus (Bx-LEC-1) were commonly detected in several proteomic analyses, demonstrating that this LEC-1 is the antigenic target of MAb-D9-F10. The localization of MAb-D9-F10 immunoreactivities at the area of the median bulb and esophageal glands suggested that the Bx-LEC-1 may be involved in food perception and digestion. The Bx-LEC-1 has two non-identical galactose-binding lectin domains important for carbohydrate binding. The affinity of the Bx-LEC-1 to D-(+)-raffinose and N-acetyllactosamine were much higher than that to L-(+)-rhamnose. Based on this combination of evidences, MAb-D9-F10 is the first identified molecular biomarker specific to the Bx-LEC-1.
Dae-Weon Lee,A Young Kim,Won Tae Kim,Kook-Jin Lim,Yeon Ho Je,Si-Hyeock Lee,Young Ho Koh 한국응용곤충학회 2008 한국응용곤충학회 학술대회논문집 Vol.2008 No.10
The pine sawyer beetle, Monochamus alternatus, transmits the pinewood nematode (PWN, Bursaphelenchus xylophilus), causing the pine wilt disease (PWD), which gives rise to enormously economic as well as forest damage. However, PWN has been identified as a pathogen of PWD, it is very difficult to discriminate B. xylophilus from B. mucronatus in a short time, which are genetically and morphologically very similar. Therefore, it has been necessary to detect and eliminate PWN-infected trees in the forest area for the prevention of PWD transmission. Up to date, there is no report on biomarkers such as DNA and protein for the diagnosis of B. xylophilus. In this study, we produced a B. xylophilus monoclonal antiserum (D9-F10) from BalbC mice and screened its specificity with various proteins extracts. Western blot analysis revealed that the D9-F10 is only reactive with B. xylophilus protein extract among other tested protein extracts, indicating that the D9-F10 is specific for a B. xylophilus protein. Furthermore, two-dimensional electrophoresis showed the D9-F10 detects a very highly acidic protein, pI≒3.5. These results suggest that the D9-F10 monoclonal antibody is useful for the development of a diagnostic kit for the pine wilt disease.
Dae-Weon Lee,A Young Kim,Won Tae Kim,Kook-Jin Lim,Yeon Ho Je,Si-Hyeock Lee,Young Ho Koh 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05
The pinewood nematode (PWN, Bursaphelenchus xylophilus) is known as a virulent factor of the pine wilt disease, transmitted to pinewoods by the pine sawyer beetle, Monochamus alternatus. It is very hard to discriminate B. xylophilus from B. mucronatus because these Bursaphelenchus species are genetically and biochemically very close. Therefore, it has been necessary to detect PWN-infected trees for the prevention of pine wilt disease transmission in a short time. We developed polyclonal antibodies against B. xylophilus in BalbC mice and primarily screened with ELISA. Positive clones releasing polyclonal antisera revealed B. xylophilus-specific immuno-reactivity, which were at least two times higher than that of B. mucronatus. Two clones, D9-F10 and 1F3, were finally selected and exhibited specific immuno-reactivity for B. xylophilus, not for B. mucronatus in Western blot analysis. D9-F10 clone was reactive with a 43-kDa whereas 1F3 clone with two proteins, 40- and 45-kDa. Their isotypes against mouse Ig family were identical, kappa-light chain. These results suggest that these monoclonal antibodies can be useful for the development of diagnostic kit for the pine wilt disease.
Dae-Weon Lee,Jong Bok Seo,Jae Soon Kang,A-YoungKim,Won Tae Kim,Yeon Ho Je,Kook Jin Lim,Si Hyeock Lee,Young Ho Koh 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10
There are increasing interests in developing methods specifically detecting pathogenic Bursaphelenchus xylophilus. In order to develop a detecting method for B .xylophilus, at first we generated monoclonal antibodies (MAbs) specific to B. xylophilus, discriminating from other pine tree resident nematodes. Among 2304 hybridoma fusions screened. We finally selected a MAb clone, 9F10 and used for further study. To identify the antigenic target of MAb-9F10, we employed several biochemical methods such as SDS-PAGE, 2 dimensional electrophoresis, anion exchange chromatography, and immunoprecipitation to separate and isolate an antigenic target. Proteins from above methods were analyzed via nano-LC-ESI-Q-IT-MS. Peptides of GaLECtin were always detected from several proteomic analyses, suggesting that GaLECtin is the antigenic target of MAb-9F10.
Ryu, Dae-Hee,Kim, Hyeock-Jin The Korean Society for Computational and Applied M 2011 Journal of applied mathematics & informatics Vol.29 No.1
We establish strong laws of large numbers for weighted sums of arrays of negatively associated random variables under the condition of h-integrability and suitable conditions on the array of weights.
류대희(Dae-Hee Ryu),이세열(Se-Yul Lee),김혁진(Hyeock-Jin Kim),송영덕(Young-Deog Song) 한국컴퓨터정보학회 2005 韓國컴퓨터情報學會論文誌 Vol.10 No.6
최근 인터넷 이용자들이 급격하게 증가하고 있으며, 초보수준의 일반 네트워크 사용자들도 인터넷상의 공개된 해킹 도구들을 사용하여 고도의 기술을 요하는 침입이 가능하여 해킹 문제가 더욱 심각해지고 있다. 해커들이 침입하기 위하여 취약점을 알아내려고 의도하는 다양한 형태의 침입시도를 사전에 탐지하여 침입이 일어나는 것을 미리 방어할 수 있는 침입시도탐지가 적극적인 예방 차원에서 더욱 필요하다. 기존의 포트 스캔이나 네트워크 취약점 공격에 대응하기 위한 네트워크 기반의 비정상 침입시도 탐지 알고리즘은 침입시도탐지에 있어 몇 가지 한계점을 갖고 있다. 기존 알고리즘은 Slow Scan, Coordinated Scan을 할 경우 탐지할 수 없다는 것이다. 따라서 침입시도 유형에 제한을 받지 않고 침입시도에 관한 다양한 형태의 비정상 접속을 효과적으로 탐지할 수 있는 새로운 개념의 알고리즘이 요구된다. 본 논문에서는 세션 패턴과 탐지 오류율을 규칙기반으로 하는 침입시도 탐지알고리즘(Session patterns & FCM Anomaly Detector : SFAD)을 제안한다. Recently, since the number of internet users is increasing rapidly and, by using the public hacking tools, general network users can intrude computer systems easily, the hacking problem is getting more serious. In order to prevent the intrusion, it is needed to detect the sign in advance of intrusion in a positive prevention by detecting the various forms of hackers intrusion trials to know the vulnerability of systems. The existing network-based anomaly detection algorithms that cope with port-scanning and the network vulnerability scans have some weakness in intrusion detection. they can not detect slow scans and coordinated scans. therefore, the new concept of algorithm is needed to detect effectively the various. In this paper, we propose a detection algorithm for session patterns and FCM.
h-STABILITY OF THE NONLINEAR PERTURBED DIFFERENCE SYSTEMS VIA n<sub>∞</sub>-SIMILARITY
Ryu, Dae Hee,Kim, Hyeock Jin,Goo, Yoon Hoe The Korean Society for Computational and Applied M 2013 Journal of applied mathematics & informatics Vol.31 No.1
In this paper, we investigate $h$-stability of the nonlinear perturbed difference system via $n_{\infty}$-similarity.