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Development of Nested-PCR Assay to Detect Acidovorax citrulli, a Causal Agent of Bacterial Fruit Blotch at Cucurbitaceae (박과 작물에 과일썩음병을 일으키는 Acidovorax citrulli 검출을 위한 nested-PCR 검사법 개발)

  • Kim, Young-Tak;Park, Kyoung-Soo;Kim, Hye-Seong;Lee, Hyok-In;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.21 no.2
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    • pp.74-81
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    • 2015
  • The specific and sensitive nested-PCR method to detect Acidovorax citrulli, a causal agent of bacterial fruit blotch on cucurbitaceae, was developed. PCR primers were designed from the draft genome sequence which was obtained with the Next Generation Sequencing of A. citrulli KACC10651, and the nested-PCR primer set (Ac-ORF 21F/Ac-ORF 21R) were selected by checking of specificity to A. citrulli with PCR assays. The selected nested-PCR primer amplified the 140 bp DNA only from A. citrulli strains, and detection sensitivity of the nested PCR increased 10,000 times of $1^{st}$ PCR detection limit (10 ng genomic DNA/PCR). The nested PCR detected A. citrulli from the all samples of seed surface wash (external seed detection) of the artificially inoculated watermelon seeds with $10^1cfu/ml$ and above population of A. citrulli while the nested PCR could not detected A. citrulli from the mashed seed suspension (internal seed detection) of the all artificially inoculated watermelon seeds. When the naturally infested watermelon seeds (10% seed infested rate with grow-out test) used, the nested PCR detected A. citrulli from 2 seed samples out of 10 replication samples externally and 5 seed samples out of 10 replication samples internally. We believe that the nested-PCR developed in this study will be useful method to detect A. citrulli from the Cucurbitaceae seeds.

Immunocytochemistry, In situ hybridization and electron microscopy for early diagnosis of Aujeszky's in living pigs (오제스키병의 생체 조기진단을 위한 면역세포화학, In situ hybridization 및 전자현미경적 연구)

  • Moon, Oun-kyong;Kim, Soon-bok;Sur, Jung-hyang;Song, Geun-suk;Nho, Whan-gook
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.845-858
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    • 1996
  • The purpose of this study was to establish early diagnostic methods for the detection of Aujeszky's disease viral antigens and nucleic acid in nasal cells, and buffy coats from experimentally infected living pigs by a combination of immunocytochemistry, in situ hybridization with digoxigenin(DIG)-labled probe and electron microscopy. Forty days old piglets were inoculated intranasally with $10^{7.0}TCID_{50}$ of Aujeszky's disease virus (ADV, NYJ-1-87 strain). The viral antigens and nucleic acid of ADV were detected in nasal cells, and buffy coat for 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopical method. The results were compared with conventional methods such as a porcine Aujeszky's disease serodiagnostic(PAD) kit, neutralization test(NT) and virus isolation. 1. The viral antigens, nucleic acids and capsids of ADV were detected in nasal cells, buffy coats from 3 days to 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopy, respectively. 2. When viral antigens were detected by the immunocytochemical technique, a diffuse brown deposit was observed in the nucleus and cytoplasm of nasal cells, buffy coats and PK-15 cells under a microscope. 3. DIG-labeled DNA probe was prepared by amplification of conserved sequence of recombinant ADV-gp50 clone with polymerase chain reacction. When ADV-DNA was detected by ISH with DIG-labeled probe, purplish blue pigmentation were observed in the nuclei and cytoplasms of ADV-infected cells under a microscope. Positive signals were observed in nasal cells and in the buffy coat and PK-15 cells at the first day after inoculation. 4. Where ADV-capsids were detected by transmission electron microscopical method, aggregation of capsids was observed in the nuclei and cytoplasms of nasal cells, buffy coats and PK-15 cells. The results suggested that these methods were considered as the highly sensitive and reliable tools for rapid and confirmative diagnosis of Aujeszky's disease in living pigs.

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Pillar Width of Twin Tunnels in Horizontal Jointed Rock Using Large Scale Model Tests (대형모형실험을 통한 수평 절리암반에서의 병설터널 이격거리)

  • Lee, Yong-Jun;Lee, Sang-Duk
    • Tunnel and Underground Space
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    • v.20 no.5
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    • pp.352-359
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    • 2010
  • Stability of twin tunnels depends on the pillar width and the ground condition. In this study, large scale model tests were conducted for investigating the influence of the pillar width of twin tunnels on their behavior in the regular horizontal jointed rock mass. Jointed rocks was composed of concrete blocks. Pillar width of twin tunnels varied in 0.29D, 0.59D, 0.88D and 1.18D, where D is the tunnel width. During the test, pillar stress, lining stress, tunnel distortion, and ground displacement were measured. Lateral earth pressure coefficient was kept in a constant value 1.0. As a result, it was found that the pillar stress and the displacement of the ground and tunnel were increased by decreasing pillar width. The maximum displacement rate was measured just after the upper excavation in each construction sequence. And the maximum influence position was the right shoulder of the preceeding tunnel at the pillar side. It was also found that for the stability assessment the inner displacement was more critical than the crown displacement. The influence zone was formed at the pillar width 0.59D~0.88D that was smaller than 0.8D~2.0D, which was proposed by experience for a good ground condition. And it would be concluded that horizontal joints could also influence on the stability of the twin tunnels.

Effect of polymerization temperature on the mechanical properties of provisional prosthesis resins (중합 온도가 임시 보철용 수지의 기계적 성질에 미치는 영향)

  • Hong, Min-Ho;Ha, Jung-Yun;Kwon, Tae-Yub
    • Korean Journal of Dental Materials
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    • v.44 no.4
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    • pp.311-318
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    • 2017
  • The purpose of this study was to examine the effects of the curing sequence and polymerization temperature on the flexural strength and microhardness of two provisional resins (Bis-acryl resin composite and polymethyl methacrylate (PMMA)). Polymerization was carried out under various conditions, in air at $25^{\circ}C$ (control) and in hot water (40, 50, 60, 70, and $80^{\circ}C$). The flexural strength test was conducted according to ISO-4049. The Knoop hardness was measured. For the Bis-acryl resin, the temperature up to $50^{\circ}C$ did not increase the flexural strength nor the hardness of the bis-acryl resin composite (p>0.05) but higher temperatures increased the strengths. For the PMMA resin, flexural strength increased with temperatures up to $70^{\circ}C$ and then decreased slightly. Bis-acryl resin composite had higher mechanical properties than the PMMA resin. The effect of heat was more pronounced in the bis-acryl resin composite than in the PMMA resin (p<0.05).

Screening of Antagonistic Bacteria Having Antifungal Activity against Brown Spot and Sheath Rot of Rice (벼 깨씨무늬병 및 잎집썩음병에 항진균 활성을 갖는 길항 미생물의 탐색)

  • Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Seo, Ji-Won;Jeong, Do-Youn
    • The Korean Journal of Mycology
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    • v.47 no.3
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    • pp.259-269
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    • 2019
  • Brown spot and sheath rot of rice are caused by fungal pathogens such as Curvularia lunata, Cochliobolus miyabeanus, and Sarocladium oryzae, and cause losses such as reduced rice yield and quality, which is an enormous problem with serious long-term effects. To search biological control agents of phytopathogenic fungi, five kinds of useful Bacillus-like isolates which are excellent in extracellular enzyme activity and produce siderophore were selected from paddy soil of Sunchang in Korea. The selected isolates were tested for excellent antifungal activity against three of the phytopathogenic fungi that frequently occur in rice, and JSRB 177 strain had the most excellent antifungal activity. Based on the experimental results, JSRB 177 is finally selected as a candidate for biological control and identified to Bacillus subtilis through 16S rRNA sequence analysis. In addition, physiological characteristics of JSRB 177 confirmed by analysis of carbohydrate fermentation patterns and enzyme production ability. Based on the above results, JSRB 177 is expected to be used as a biological control agent for the rice pathogenic fungi. In the future, further studies related to industrialization such as port test and establishment of mass production process are needed.

Identification and Chemical Control of Gray Snow Molds Caused by Typhula spp. on Golf Course in Korea (우리나라의 골프코스에서 Typhula spp.에 의해 발생하는 설부병의 동정 및 방제)

  • Kim, Jeong-Ho;Shim, Gyu-Yul;Lee, Hye-Min;Moon, Hyo-Sun;Kim, Young-Ho
    • Asian Journal of Turfgrass Science
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    • v.21 no.2
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    • pp.147-154
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    • 2007
  • In March of 2004, gray snow mold (Typhula blight) caused by Typhula spp. occurred on perennial ryegrass (Lolium perenne L.) and Kentucky bluegrass (Poo pratensis L.) at MuJu golf courses in Jeonbuk Province. Leaves in the affected areas were matted together and frequently covered with white to grayish mycelia. Sclerotia were formed on the leaf blade, leaf sheath, or crown regions. The fungus isolated from the diseased leaf formed whitish mycelium, clamp connections, and light pink to brown, irregular-shaped small sclerotia of less than 1.4 mm in diameter, which are characteristic to Typhula incarnata. Optimum temperature ranges for mycelial growth were $5^{\circ}C$ to $15^{\circ}C$. The causal organism was confirmed to be T. incarnata as the partial sequence of its ribosomal RNA ITS1 (internal transcribed spacer) region was 91% homologous to those of T. incarnata in GenBank database. Out of the 14 fungicides tested fur antifungal activity in vitro, 10 fungicides including iprodione, tebuconazole, polyoxin D, flutolanil, hexaconazole, tolclofos-methyl, fosetyl-Al, mepronil, pencycuron+tebuconazole, and fenarimol completely inhibited fungal growth at their recommended concentrations. In the field test, these fungicides and others such as thifluzamide and thiram effectively controlled the gray snow mold of turfgrass with some variable degrees of control efficacies.

Molecular phylogenetic relationships and speciation of Ranunculus cantoniensis (Ranunculaceae) (털개구리미나리(Ranunculus cantoniensis)의 분자계통학적 유연관계 및 종분화)

  • Lee, Chang Shook;Lee, Nam Sook;Yeau, Sung Hee
    • Korean Journal of Plant Taxonomy
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    • v.34 no.4
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    • pp.335-358
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    • 2004
  • To investigate molecular phylogenetic relationships and to test hypothesis of hybrid origin of Ranunculus cantoniensis (Ranunculaceae), the sequences of nrDNA and chloroplast DNA were analyzed for 8 taxa and 25 accessions including 5 accessions of outgroup. In the phylogenetic trees by analyses of maximum parsimony and maximum likelihood for ITS nrDNA sequences and combined data of psbA-trnH, rps16 and trnL sequences of cpDNA, R. cantoniensis was most closely related to R. chinensis, and then to R. taciroi and R. silerifolius. The molecular phylogenetic relationships were not congruent with the previous report that R. cantoniensis was most closely related to R. silerifolius. In the sequence analysis of ITS and psbA-trnH, rps16, trnL for R. cantoniensis and the related taxa, R. cantoniensis showed polymorphism. It supported that the polymorphism also was reported in chromosome number and karyotype of R. cantoniensis. Ranunculus cantoniensis shared the marker gene of R. chinensis and R. silerifolius in ITS, and one of R. silerifolius in cpDNA. These results supported the hypothesis that R. cantoniensis was caused by hybridization between R. chinensis and R. silerifolius based on chromosome number and karyotype, and also estimated that R. silerifolius might be of maternal origin and R. chinensis be paternal.

Genetic Variation of Pinus densiflora Populations in South Korea Based on ESTP Markers (ESTP 표지를 이용한 국내 소나무 집단의 유전변이)

  • Ahn, Ji Young;Hong, Kyung Nak;Lee, Jei Wan;Hong, Yong Pyo;Kang, Hoduck
    • Korean Journal of Plant Resources
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    • v.28 no.2
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    • pp.279-289
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    • 2015
  • Genetic diversity and genetic differentiation of thirteen Pinus densiflora populations in South Korea were estimated using nine ESTP (Expressed Sequence Tag Polymorphism) markers. The numbers of allele and the effective allele were 2.2 and 1.8, respectively. The percentage of polymorphic loci (P) was 98.8%. The observed and the expected heterozygosity were 0.391 and 0.402, respectively, and the eleven populations except for Ahngang and Gangneung population were under Hardy-Weinberg equilibrium state. The level of genetic differentiation (Wright’s FST = 0.057) was higher than those of isozyme or nSSR markers. We could not find out any relationship between the genetic distance and geographic distribution among populations from cluster analysis. Also, the genetic differentiation between populations was not correlated with the geographic distance (r = 0.017 and P = 0.344 from Mantel test). From the result of FST-outlier analysis to identify a locus under selection, six loci were detected at confidence interval of 99% by the frequentist’s method. However, only three loci (sams2+AluⅠ, sams2+RsaⅠ, PtNCS_p14A9+HaeⅢ) were presumed as outliers by Bayesian method. The sams2+AluⅠ and sams2+RsaⅠlocus were originated from the sams2 gene and seemed to be the loci under balancing selection.

Design of Fetal Health Classification Model for Hospital Operation Management (효율적인 병원보건관리를 위한 태아건강분류 모델)

  • Chun, Je-Ran
    • Journal of Digital Convergence
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    • v.19 no.5
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    • pp.263-268
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    • 2021
  • The purpose of this study was to propose a model which is suitable for the actual delivery system by designing a fetal delivery hospital operation management and fetal health classification model. The number of deaths during childbirth is similar to the number of maternal mortality rate of 295,000 as of 2017. Among those numbers, 94% of deaths are preventable in most cases. Therefore, in this paper, we proposed a model that predicts the health condition of the fetus using data like heart rate of fetuses, fetal movements, uterine contractions, etc. that are extracted from the Cardiotocograms(CTG) test using a random forest. If the redundancy of the data is unbalanced, This proposed model guarantees a stable management of the fetal delivery health management system. To secure the accuracy of the fetal delivery health management system, we remove the outlier which embedded in the system, by setting thresholds for the upper and lower standard deviations. In addition, as the proportion of the sequence class uses the health status of fetus, a small number of classes were replicated by data-resampling to balance the classes. We had the 4~5% improvement and as the result we reached the accuracy of 97.75%. It is expected that the developed model will contribute to prevent death and effective fetal health management, also disease prevention by predicting and managing the fetus'deaths and diseases accurately in advance.

An Interpretable Log Anomaly System Using Bayesian Probability and Closed Sequence Pattern Mining (베이지안 확률 및 폐쇄 순차패턴 마이닝 방식을 이용한 설명가능한 로그 이상탐지 시스템)

  • Yun, Jiyoung;Shin, Gun-Yoon;Kim, Dong-Wook;Kim, Sang-Soo;Han, Myung-Mook
    • Journal of Internet Computing and Services
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    • v.22 no.2
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    • pp.77-87
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    • 2021
  • With the development of the Internet and personal computers, various and complex attacks begin to emerge. As the attacks become more complex, signature-based detection become difficult. It leads to the research on behavior-based log anomaly detection. Recent work utilizes deep learning to learn the order and it shows good performance. Despite its good performance, it does not provide any explanation for prediction. The lack of explanation can occur difficulty of finding contamination of data or the vulnerability of the model itself. As a result, the users lose their reliability of the model. To address this problem, this work proposes an explainable log anomaly detection system. In this study, log parsing is the first to proceed. Afterward, sequential rules are extracted by Bayesian posterior probability. As a result, the "If condition then results, post-probability" type rule set is extracted. If the sample is matched to the ruleset, it is normal, otherwise, it is an anomaly. We utilize HDFS datasets for the experiment, resulting in F1score 92.7% in test dataset.