• Title/Summary/Keyword: Tool-chain

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Antibiotic Resistance and Bacterial Biofilm Formation by Staphylococcus aureus Strains Isolated from Various Foods (식품에서 분리한 황색포도상구균의 항생제 내성 특징 및 균막 형성)

  • Lee, Joo-Young;Wang, Hae-Jin;Shin, Dong-Bin;Cho, Yong-Sun
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.96-104
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    • 2013
  • Staphylococcus aureus is a major human pathogen that produces a wide array of toxins, leading to a number of adverse symptoms. We examined 275 strains of Staphylococcus aureus isolated from various foods between 2006 and 2008 for antimicrobial susceptibility. At least 259 (94.2%) of the tested strains showed antibiotic resistant properties, and 106 (40.7%) of them showed multiple antibiotic resistance. Eleven of the tested strains were resistant to oxacillin and mec A-positive. Moreover, oxacillin-resistant strains were significantly more likely to be multi-drug resistant (p < 0.01). Of the 275 isolates tested, 24.4% were noted as being positive for slime production and 30.5% were positive for biofilm assay. Antibiotic resistance was not associated with a significantly higher prevalence of biofilm formation. Twenty strains were classified using the DiversiLab system. Most of the strains could be classified into 2 clusters and 4 unique types. All 10 mec A-positive strains (cluster I) were grouped together into the same sub-cluster. Cluster II (6 strains) was not found to be resistant to oxacillin in this study. Although the prevalence of methicillin-resistant S. aureus in food is currently low, the risk of its transmission through the food chain cannot be disregarded.

Arsenic Concentrations of Groundwater and Rice Grains in Bangladesh and Phytoremediation (방글라데시의 지하수와 쌀의 비소오염 및 식물정화법)

  • Islam, Jahidul Mohammad;Kim, Bomchul;Laiju, Nahida;Nasirullah, Tarek;Miah, Mohammad Nuruddin
    • Journal of Korean Society on Water Environment
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    • v.26 no.1
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    • pp.116-124
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    • 2010
  • While groundwater is the major source for drinking and irrigation purposes, arsenic (As) contamination of groundwater is a serious issue in Bangladesh. With a view to reduce As contamination in drinking water the guideline value recommended for Bangladesh is 0.05 mg/L. We assessed groundwater As in an As-affected Sadar Upazilla (small administrative unit) in the District (administrative unit) of Chapai Nabwabganj during 2006, where 50% hand tube well water were above the recommended limit (0.05 mg/L) during dry season. Almost 20% tube well waters were above the recommended limit during rainy season, perhaps due to the dilution of water table. The groundwater in Bangladesh contaminates surface soils and plants thereby As entering the food chain. In 2005, we examined the As levels in different rice varieties grown in different Districts of Bangladesh and the As concentrations in rice grain ranged from 0.07~1.12 mg/kg while the concentrations in 3 rice varieties were above the recommended limit (1 mg/kg rice grain) and the maximum concentration was 1.12 mg/kg rice grain in the rice variety BR 11. With few exceptions, the As content of rice grain in Bangladesh is not considered to be concentration of greater health concern as yet. We also observed enhanced root uptake, efficient root-to shoot translocation, and a much elevated tolerance through internal detoxification all contribute to As hyperaccumulation in a plant, ladder brake fern (Pteris vittata L.). But the phytoremediation technique might not be an appropriate tool to reduce the As calamity in the vast areas of Bangladesh. To mitigate the As problem of Bangladesh, better coordination among governmental agencies and many other organizations will be required to combat the disaster.

Expression of Human p53 Gene as Glutathione S-transferase Fusion Proteins in Escherichia coli (사람의 p53 유전자와 Glutathione S-Transferase와의 융합 단백질의 대장균에서의 발현)

  • 오상진
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.279-285
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    • 1993
  • Alterations of the p53 gene arc among the most frequent genetic changes in human cancer and often result in increased levels of p53 protein within the malignant cells. Detection of accumulated p53 protein can be a useful prognostic tool in human cancer. In order to make polyclonal antibodies for immunohistochemical screening. human p53 gene was expressed in E. coli in the form of GST (glutathione S-transfi.:rase) fusion proteins. Two p53 gene fragments. which were N('()I small fragment encoding amino acid residues of 1-151-: and Ncol large fragment of 159-393. were subeloned into the unique BamHI site present within the pGEX-2T vector using BamHI linker and recombinant plasmids pGTNS and pGTNL were constructed. respectively. The p53 cDNA fragment (from pC53-$SN_3$,) encoding amino acid 38-145 (proline at residue 72) was amplified by polymerase chain reaction(PCR). The amplified DNA was digested with BamHI and Prull and inserted into the BamHI-Smal sites of pG EX-2T and recombinant plasmid pGTBP was constructed. After IPTG induction of these plasmids for 4 hours. fusion proteins were purified from E. coli extracts with glutathione Sepharose beads. The bound proteins were resolved by 10% SDS-polyacrylamide gel electrophoresis and the molecular weights were 54 kDa. 53 kDa and 40 kDa. respectively. Approximately one milligram of fusion proteins were purified from 1 -liter cultures.

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Development of a Multiplex PCR Assay for Rapid Identification of Larimichthys polyactis, L. crocea, Atrobucca nibe, and Pseudotolithus elongates (다중 PCR 분석법을 이용한 참조기, 부세, 흑조기 및 긴가이석태의 신속한 종판별법 개발)

  • Noh, Eun Soo;Lee, Mi-Nan;Kim, Eun-Mi;Park, Jung Youn;Noh, Jae Koo;An, Cheul Min;Kang, Jung-Ha
    • Journal of Life Science
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    • v.27 no.7
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    • pp.746-753
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    • 2017
  • In order to rapidly identify four drums species, Larimichthys polyactis, L. crocea, Atrobucca nibe, and Pseudotolithus elongates, a highly efficient and quick method has been developed using multiplex polymerase chain reaction (PCR) with species-specific primers. Around 1.4 kbp of the mitochondrial COI gene sequences from the four drums species were aligned, and species-specific forward primers were designed, based on the single nucleotide polymorphism (SNP). The optimal conditions for PCR amplification were selected through cross-reactivity, using a gradient PCR method. The PCR results demonstrated species-specific amplification for each species at annealing temperatures between 50 and $62^{\circ}C$. Multiplex species-specific PCR (MSS-PCR) amplification reactions with four pairs of primers were performed for sixteen specimens of each species. MSS-PCR lead to a species-specific amplification of a 1,540 bp fragment in L. polyactis, 1,013 bp in A. nibe, 474 bp in L. crocea, and 182 bp in P. elongates, respectively. The four different sizes of each PCR product can be quickly and easily detected by single gel electrophoresis. The sensitivity of the MSS-PCR of the DNA was up to $0.1ng/{\mu}l$ as a starting concentration for the four different species tested. These results suggest that MSS-PCR, with species-specific primers based on SNP, can be a powerful tool in the rapid identification of the four drums species, L. polyactis, L. crocea, A. nibe, and P. elongates.

An Optimized V&V Methodology to Improve Quality for Safety-Critical Software of Nuclear Power Plant (원전 안전-필수 소프트웨어의 품질향상을 위한 최적화된 확인 및 검증 방안)

  • Koo, Seo-Ryong;Yoo, Yeong-Jae
    • Journal of the Korea Society for Simulation
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    • v.24 no.4
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    • pp.1-9
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    • 2015
  • As the use of software is more wider in the safety-critical nuclear fields, so study to improve safety and quality of the software has been actively carried out for more than the past decade. In the nuclear power plant, nuclear man-machine interface systems (MMIS) performs the function of the brain and neural networks of human and consists of fully digitalized equipments. Therefore, errors in the software for nuclear MMIS may occur an abnormal operation of nuclear power plant, can result in economic loss due to the consequential trip of the nuclear power plant. Verification and validation (V&V) is a software-engineering discipline that helps to build quality into software, and the nuclear industry has been defined by laws and regulations to implement and adhere to a through verification and validation activities along the software lifecycle. V&V is a collection of analysis and testing activities across the full lifecycle and complements the efforts of other quality-engineering functions. This study propose a methodology based on V&V activities and related tool-chain to improve quality for software in the nuclear power plant. The optimized methodology consists of a document evaluation, requirement traceability, source code review, and software testing. The proposed methodology has been applied and approved to the real MMIS project for Shin-Hanul units 1&2.

Analysis of Genomic Structure of an Aflatoxin Biosynthesis Homologous Gene Cluster in Aspergillus oryzae RIB Strains

  • Lee, Yun-Hae;Tominaga, Mihoko;Hayashi, Risa;Sakamoto, Kazutoshi;Yamada, Osamu;Akita, Osamu
    • 한국균학회소식:학술대회논문집
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    • 2006.10a
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    • pp.32-44
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    • 2006
  • To investigate non-aflatoxin-production of A. oryzae at the molecular level, an aflatoxin biosynthesis gene homolog cluster of RIB 40 was analyzed. Although most genes in the corresponding cluster exhibited from 97 to 99 % similarity to those of Aspergillus flavus, three genes shared 93 % similarity or less. In addition, although slight expression of aflR, positive transcriptional regulator gene, was detected in some A. oryzae strains having seven aflatoxin biosynthesis homologous genes, other genes related to aflatoxin production were not detected. RIB strains were mainly divided into group 1, having seven aflatoxin biosynthesis homologous genes (aflT, nor-i, aflR, norA, avnA, verB, and vbs), and group 2, having three homologous (avnA, verB, and vbs). Partial aflatoxin homologous gene cluster of RIB62 from group 2 was sequenced and compared with that of RIB40 from group 1. RIB62 showed a large deletion upstream of ver-1 with more than half of the aflatoxin homologous gene cluster missing including aflR, a positive transcriptional regulatory gene. Adjacent to the deletion of the aflatoxin homologous gene cluster, RIB62 has a unique sequence of about 8kb and a telomere. Southern analysis of A. oryzae RIB strains with four kinds of probe derived from the unique sequence of RIB62 showed that all group 2 strains have identical hybridizing signals. Polymerase chain reaction with specific primer set designed to amplify the junction between ver-1 and the unique sequence of RIB62 resulted in the same size of DNA fragment only from group 2 strains. Based on these results, we developed a useful genetic tool that distinguishes A. oryzae group 2 strains from the other groups' strains and propose that it might have differentiated from the ancestral strains due to chromosomal breakage.

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Usefulness of Automated PCR Test for the Detection of Mycobacterium Tuberculosis in Fresh Biopsy Tissues (신선조직 검체에서 결핵균 검출을 위한 자동화 중합효소연쇄반응 검사의 유용성)

  • Choi, Woo Soon;Shin, So Young;Kim, Jong Ok;Kim, Myung Sook;Lee, Hye Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.61 no.1
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    • pp.54-59
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    • 2006
  • Background: Although there have been several studies regarding the clinical value of an automated TB-PCR study using sputum, bronchial washing, and other body fluid samples for the detection of pulmonary tuberculosis, there are only a few reports on the use of fresh tissue samples. Materials and methods: The acid-fast bacilli stain(AFB), tuberculosis culture, automated TB-PCR study, and histopathology examination were performed in 42 fresh tissue samples. Results: Among the 42 cases, 18 cases were diagnosed with tuberculosis based on the clinical findings. Sixteen of the 18 cases were TB-PCR positive and of these 16 cases, only 2 cases were positive in the AFB stain or culture study. However, all 18 cases showed the histopathology findings of chronic granulomatous inflammation that was compatible with tuberculosis. Based on the clinical findings, the sensitivity, specificity, positive predictability, and negative predictability of the automated TB-PCR study were 88.9%, 100%, 100%, and 92.3% respectively. Conclusion: An automated TB-PCR assay is an important diagnostic tool for diagnosing tuberculosis in fresh tissue samples.

Development of a multiplex qRT-PCR assay for detection of African swine fever virus, classical swine fever virus and porcine reproductive and respiratory syndrome virus

  • Chen, Yating;Shi, Kaichuang;Liu, Huixin;Yin, Yanwen;Zhao, Jing;Long, Feng;Lu, Wenjun;Si, Hongbin
    • Journal of Veterinary Science
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    • v.22 no.6
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    • pp.87.1-87.12
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    • 2021
  • Background: African swine fever virus (ASFV), classical swine fever virus (CSFV), and porcine reproductive and respiratory syndrome virus (PRRSV) are still prevalent in many regions of China. Co-infections make it difficult to distinguish their clinical symptoms and pathological changes. Therefore, a rapid and specific method is needed for the differential detection of these pathogens. Objectives: The aim of this study was to develop a multiplex real-time quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) for the simultaneous differential detection of ASFV, CSFV, and PRRSV. Methods: Three pairs of primers and TaqMan probes targeting the ASFV p72 gene, CSFV 5' untranslated region, and PRRSV ORF7 gene were designed. After optimizing the reaction conditions, including the annealing temperature, primer concentration, and probe concentration, multiplex qRT-PCR for simultaneous and differential detection of ASFV, CSFV, and PRRSV was developed. Subsequently, 1,143 clinical samples were detected to verify the practicality of the assay. Results: The multiplex qRT-PCR assay could specifically and simultaneously detect the ASFV, CSFV, and PRRSV with a detection limit of 1.78 × 100 copies for the ASFV, CSFV, and PRRSV, but could not amplify the other major porcine viruses, such as pseudorabies virus, porcine circovirus type 1 (PCV1), PCV2, PCV3, foot-and-mouth disease virus, porcine parvovirus, atypical porcine pestivirus, and Senecavirus A. The assay had good repeatability with coefficients of variation of intra- and inter-assay of less than 1.2%. Finally, the assay was used to detect 1,143 clinical samples to evaluate its practicality in the field. The positive rates of ASFV, CSFV, and PRRSV were 25.63%, 9.36%, and 17.50%, respectively. The co-infection rates of ASFV+CSFV, ASFV+PRRSV, CSFV+PRRSV, and ASFV+CSFV+PRRSV were 2.45%, 2.36%, 1.57%, and 0.17%, respectively. Conclusions: The multiplex qRT-PCR developed in this study could provide a rapid, sensitive, specific diagnostic tool for the simultaneous and differential detection of ASFV, CSFV, and PRRSV.

Development of ISSR-Derived SCAR Markers for Identification of Jujube Cultivars (대추나무 품종 식별을 위한 ISSR 유래 SCAR 표지 개발)

  • Nam, Jae-Ik;Kim, Chul-Woo;Kim, Sea-Hyun
    • Journal of Korean Society of Forest Science
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    • v.108 no.3
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    • pp.302-310
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    • 2019
  • Precise and fast identification of crop cultivars is essential for efficient breeding and plant breeders' rights. Traditional methods for identification of jujube cultivars are based on the evaluation of morphological characteristics. However, due to time constraints and environmental influences, it is difficult to distinguish cultivars using only morphological traits. In this study, we cloned fragments from improved inter simple sequence repeats (ISSR) analysis, and developed stably diagnostic sequence-characterized amplified region (SCAR) markers. The specific ISSR bands of jujube cultivars from Dalizao and Boeundaechu were purified, cloned, and sequenced. As a result, four clones labeled 827Dalizao550, 827Boeun750, 846Boeun700, and 847Dalizao850 were identified. In order to investigate whether they were specific for the jujube cultivar, four pairs of SCAR primers were then designed and polymerase chain reaction (PCR) amplifications were conducted to analyze 32 samples, including jujube and sour jujube. In the PCR amplification of the 827Dalizao550 SCAR marker, the specific bands with 550 bp were amplified in six samples (Dalizao, Sandonglizao, Dongzao, Yuanlin No. 2, Suanzao 2, Suanzao 4), but unexpected bands (490 bp) were amplified in the others. Moreover, in the PCR amplification of the 847Dalizao850 SCAR marker, the specific bands with 850 bp were found in three samples (Dalizao, Sandonglizao, and Dongzao) and 900 bp unexpected bands were amplified in five samples (Pozao, Suanzao 1, Suanzao 2, Suanzao 3, Suanzao 4). These results showed that newly developed markers could be useful as a fast and reliable tool to identify jujube cultivars. However, further identification of polymorphic information and the development of SCAR markers are required for the identification of more diverse cultivars.

Classification of Trusted Boot Technology Components based on Hardware Dependency (하드웨어 종속/독립성에 따른 신뢰성 부팅 기술 구성 요소 분류)

  • Park, Keon-Ho;Kim, Sieun;Lee, Yangjae;Lee, SeongKee;Kang, Tae In;Kim, Hoon Kyu;Park, Ki-woong
    • The Journal of Korean Institute of Next Generation Computing
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    • v.14 no.6
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    • pp.44-56
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    • 2018
  • Researches on military weapons are actively studied to improve national defense power of each country. The military weapon system is being used not only as a weapon but also as a reconnaissance and surveillance device for places where it is difficult for people to access. If such a weapon system becomes an object of attack, military data that is important to national security can be leaked. Furthermore, if a device is taken, it can be used as a terrorist tool to threaten its own country. So, security of military devices is necessarily required. In order to enhance the security of a weapon system such as drone, it is necessary to form a chain of trust(CoT) that gives trustworthiness to the overall process of the system from the power on until application is executed. In this paper, by analyzing the trusted computing-based boot technology, we derive trusted boot technology components and classify them based on hardware dependence/independence. We expect our classification of hardware dependence/independence to be applied to the trusted boot technology of our self-development ultraprecision weapon system to improve the defense capability in our military.