• Title/Summary/Keyword: and variety identification

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MAPK Activation and Cell Viability after $H_2O_2$ Stimulation in Cultured Feline Ileal Smooth Muscle Cells

  • Song, Hyun-Ju;Jeong, Ji-Hoon;Lee, Dong-Kyu;Lee, Tai-Sang;Min, Young-Sil;Sohn, Uy-Dong
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.6
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    • pp.339-344
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    • 2004
  • Recent data have shown the importance of oxidative stresses in the pathogenesis of inflammatory bowel disease, crohn's disease and ulcerative colitis. $H_2O_2$, reactive oxygen species (ROS) donor, has been reported to act as a signaling molecule involved in a variety of cellular functions such as apo/ptosis and proliferation. In the present study, we investigated viability of cultured ileal smooth muscle cells (ISMC) after stimulation with $H_2O_2$. Trypan blue method revealed that the cell viability of ISMC treated with 1 mM $H_2O_2$ was not different from that of controls at up to 2 h time point, while treatment of ISMC with 1 mM $H_2O_2$ for 48 h finally induced significant decrease in the cell viability. Therefore, we evaluated whether $H_2O_2$ was capable of ERKs activation in ISMC for the short-term exposure and examined whether tyrosine kinase was involved in the process of ERK activation by $H_2O_2$ in ISMC. We also investigated the effects of $H_2O_2$ on activation of SAPK/JNK and p38 MAP kinase in ISMC. Thus, ISMC were cultured and exposed to $H_2O_2$, and western blot analysis was performed with phosphospecific MAP kinase antibodies. Robust activation of ERK occurred within 30 min of 1 mM $H_2O_2$ treatment. $H_2O_2-induced$ ERK activation was attenuated by a tyrosine kinase inhibitor, genistein, indicating that tyrosine kinase was probably involved in the ERK activation by $H_2O_2$. $H_2O_2$ was a moderate activator of SAPK/JNK, while p38 MAP kinase was not activated by $H_2O_2$. We suggest that ERK activation induced by short-term $H_2O_2$ treatment plays a critical role in cellular protection in the early stage of response to oxidative stress. The present study suggests the necessity of identification of MAPK signaling pathways affected by ROS, since it could ultimately elucidate cellular consequences involved in initiation and perpetuation of intestinal tissue damage in the diseases such as crohn's disease and ulcerative colitis, resulted from excessive ROS.

MLPA Applications in Genetic Testing (유전자진단에 있어서 Multiplex Ligation Dependent Probe Amplification (MLPA)의 이론과 실제)

  • Kim, Gu-Hwan;Lee, Beom-Hee;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.6 no.2
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    • pp.146-154
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    • 2009
  • Multiplex ligation dependent probe amplification (MLPA) is a PCR-based method to detect gene dosage. Since its introduction, MLPA has been used to test a large number of genes for major deletions or duplications. Genetic testing, as a diagnostic tool for genetic disease, has been used primarily to identify point mutations, including base substitutions and small insertions/deletions, using PCR and sequence analysis. However, it is difficult to identify large deletions or duplications using routine PCR- gel based assays, especially in heterozygotes. The MLPA is a more feasible method for identification of gene dosage than another routine PCR-based methods, and better able to detect deleterious deletions or duplications. In addition to detection of gene dosage, MLPA can be applied to identify methylation patterns of target genes, aneuploidy during prenatal diagnoses, and large deletions or duplications that may be associated with various cancers. The MLPA method offers numerous advantages, as it requires only a small amount of template DNA, is applicable to a wide variety of applications, and is high-throughput. On the other hand, this method suffers from disadvantages including the possibility of false positive results affected by template DNA quality, difficulties identifying SNPs located in probe sequences, and analytical complications in quantitative aspects.

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A Measurement System for Color Environment-based Human Body Reaction (색채 환경 기반의 인체 반응 정보 측정 시스템)

  • Kim, Ji-Eon;Jeong, Chang-Won;Joo, Su-Chong
    • Journal of Internet Computing and Services
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    • v.17 no.2
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    • pp.59-65
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    • 2016
  • The result of analyzing the cognitive reaction due to the color environment has been applied to various filed especially in medical field. Moreover, the study about the identification of patient's condition and examination the brain activity by collecting the bio-signal based on the color environment is being actively conducted. Even though, there were a variety of experiments by convention the color environment using a light or LED color, it still has a problem that affects the psychological information. Therefore, our proposed system using a HMD (Head Mounting display) to provide a completed color environment condition. This system uses the BMS(Biomedical System) to collect the biometric information which responds to the specific color condition and the human body response information can be measured by the development the Memory and Attention test on Mobile phone. The collection of Biometric information includes electro cardiogram(ECG), respiration, oxygen saturation (Sp02), Bio-impedance, blood pressure will store in the database. In addition, we can verify the result of the human body reaction in the color environment by Memory and Attention application. By utilizing the reaction of the human body information that is collected thought the proposed system, we can analyze the correlation between the physiological information and the color environment. And we also expect that this system can apply to the medical diagnosis and treatment. For future work, we will expand the system for prediction and treatment of Alzheimer disease by analyzing the visualization data through the proposed system. We will also do evaluation on the effectiveness of the system for using in the rehabilitation program.

Characterization of a Restriction Endonuclease AspJI from Alcaligenes sp. J-482 (Alcaligenes sp. J-482 로부터 분리한 제한효소 AspJI의 특성)

  • Lee, Jeong-Taek;;Lim, Jai-Yun
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.285-290
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    • 1994
  • About 500 bacterial and fungal strains from a wide variety of natural habitats were screened for a new type II restriction endonuclease. Among the 500 species, we selected one species that produced a new restriction endonuclease. This strain has an optimum temperature of $30^{circ}C$ for growth. Morphological, cultural, and physiological characteristics were examined for identification of the isolated strain J-482. This strain was found to belong to the genus Alcaligenes. The restriction endonuclease was named as AspJI and partially purified from Alcaligenes sp. J-482 by DEAE-Sephadex A-50 column chromatography and gel filtration. Most of other nucleases were removed by the purification steps. The AspJI has a substrate specificity to ${lambda}$ DNA, pBR322 and Adenovirus-2 DNA. For its maximal activity, the isolated enzyme requires $MgCl_2$, which should be at least 12.5 mM and it does not need any other cofactors. It is maximally active in the absence of NaCl and is completely inactivated at 100 mM NaCl. The pH and temperature optima for activity were pH 7.5 and $37^{circ}C$, respectively. The DNA fragments generated by digesting ${lambda}$ DNA, pBR322, and Adenovirus-2 DNA with AspJI were the same as that produced by AatII. This suggests that AspJI is an isoschizomer of AatII.

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Identification of Novel Bacillus subtilis IDCC 9204 Producing a High-Level Fibrinolytic Enzyme and Properties of NK-IL9204 (고농도 혈전용해효소를 생산하는 신규 Bacillus subtilis IDCC 9204의 분리 및 NK-IL9204의 효소학적 특성)

  • Lee, Seung-Hun;An, Gwangmin;Kim, Heu-Hang;Kang, Jae-Hoon;Kang, Dae-Jung
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.600-606
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    • 2012
  • A Bacillus sp. that produces fibrinolytic enzyme was isolated from Cheonggukjang, a traditional Korean soybean-fermented food. According to 16S rRNA gene base sequencing, the bacillus was identified as a variety of Bacillus subtilis, and named Bacillus subtilis IDCC 9204. Fibrinolytic enzyme NK-IL9204 was stable up to $60^{\circ}C$ and within pH range of 5-10. Purified NK-IL9204 was detected through fibrin zymography. The molecular weight and isoelectric point of the enzyme were estimated to be 27.7 kDa and 6.7 by SDS-PAGE and 2D electrophoresis, respectively. Its amino acid sequence was similar to that of nattokinase (identities 99.5%) and different from that of nattokinase BPN (identities 86.4%). The plasma fibrinolytic activity of NK-IL9204 was measured by euglobulin clot lysis times (ECLT). The NK-IL9204 was orally administered to SD rats for 3 weeks (1,000 FU/rat/day). The ECLT was significantly shortened by supplementation of NK-IL9204.

Antimicrobial Susceptibility and Biochemical Characteristics of Streptococcus suis Isolated from Diseased Pigs in Gyeongbuk Province (경북지방 환돈에서 분리한 Streptococcus suis의 생화학적 성상 및 약제감수성)

  • Choi, Seong-Kyoon;Kim, Seong-Guk;Kim, Young-Hoan;Choi, Jeong-Hye;Jo, Min-Hee;Cho, Gil-Jae
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1549-1555
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    • 2010
  • Streptococcus suis is a worldwide pathogen of a variety of porcine infection and has also been described as a pathogen for humans. We studied biochemical characteristics, antimicrobial susceptibility, and identification of polymerase chain reaction (PCR) of S. suis isolated from diseased pigs in Gyeongbuk province from 2004 to 2009. Sixty-one isolates were identified as S. suis by biochemical characteristics and PCR from 40 farms. The biochemical characteristics of S. suis isolates were production of VP-negative, hippurate, esculin, and arginine decarboxylase-positive, and fermentation of carbohydrate was variable lactose, trehalose, inulin, and raffinose, which was typeable 11 phenotype. In an antimicrobial susceptibility test, the majority of isolates were highly susceptible to amoxicillin/clavulanic acid, ampicillin, cephalothin, cefoperazone and florfenicol, while being highly resistant to streptomycin, kanamycin, amikacin, neomycin, erythromycin, clindamycin, and tetracycline. The isolates were divided into 11 phenotypes of biochemistry. By using PCR, the 16S-rRNA gene DNA fragment was detected at 304 bp from all of isolates. These results may provide the basic information needed to establish strategies for the prevention of S. suis infection in pigs.

Field Phenotyping of Plant Height in Kenaf (Hibiscus cannabinus L.) using UAV Imagery (드론 영상을 이용한 케나프(Hibiscus cannabinus L.) 작물 높이의 노지 표현형 분석)

  • Gyujin Jang;Jaeyoung Kim;Dongwook Kim;Yong Suk Chung;Hak-Jin Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.67 no.4
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    • pp.274-284
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    • 2022
  • To use kenaf (Hibiscus cannabinus L.) as a fiber and livestock feed, a high-yielding variety needs to be identified. For this, accurate phenotyping of plant height is required for this breeding purpose due to the strong relationship between plant height and yield. Plant height can be estimated using RGB images from unmanned aerial vehicles (UAV-RGB) and photogrammetry based on Structure from Motion (SfM) algorithms. In kenaf, accurate measurement of height is limited because kenaf stems have high flexibility and its height is easily affected by wind, growing up to 3 ~ 4 m. Therefore, we aimed to identify a method suitable for the accurate estimation of plant height of kenaf and investigate the feasibility of using the UAV-RGB-derived plant height map. Height estimation derived from UAV-RGB was improved using multi-point calibration against the five different wooden structures with known heights (30, 60, 90, 120, and 150 cm). Using the proposed method, we analyzed the variation in temporal height of 23 kenaf cultivars. Our results demontrated that the actual and estimated heights were reliably comparable with the coefficient of determination (R2) of 0.80 and a slope of 0.94. This method enabled the effective identification of cultivars with significantly different heights at each growth stages.

Gene Expression Profiling in Diethylnitrosamine Treated Mouse Liver: From Pathological Data to Microarray Analysis (Diethylnitrosamine 처리 후 병리학적 결과를 기초로 한 마우스 간에서의 유전자 발현 분석)

  • Kim, Ji-Young;Yoon, Seok-Joo;Park, Han-Jin;Kim, Yong-Bum;Cho, Jae-Woo;Koh, Woo-Suk;Lee, Michael
    • Toxicological Research
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    • v.23 no.1
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    • pp.55-63
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    • 2007
  • Diethylnitrosamine (DEN) is a nitrosamine compound that can induce a variety of liver lesions including hepatic carcinoma, forming DNA-carcinogen adducts. In the present study, microarray analyses were performed with Affymetrix Murine Genome 430A Array in order to identify the gene-expression profiles for DEN and to provide valuable information for the evaluation of potential hepatotoxicity. C57BL/6NCrj mice were orally administered once with DEN at doses of 0, 3, 7 and 20 mg/kg. Liver from each animal was removed 2, 4, 8 and 24 hrs after the administration. The histopathological analysis and serum biochemical analysis showed no significant difference in DEN-treated groups compared to control group. Conversely, the principal component analysis (PCA) profiles demonstrated that a specific normal gene expression profile in control groups differed clearly from the expression profiles of DEN-treated groups. Within groups, a little variance was found between individuals. Student's t-test on the results obtained from triplicate hybridizations was performed to identify those genes with statistically significant changes in the expression. Statistical analysis revealed that 11 genes were significantly downregulated and 28 genes were upregulated in all three animals after 2 h treatment at 20 mg/kg. The upregulated group included genes encoding Gdf15, JunD1, and Mdm2, while the genes including Sox6, Shmt2, and SIc6a6 were largely down regulated. Hierarchical clustering of gene expression also allowed the identification of functionally related clusters that encode proteins related to metabolism, and MAPK signaling pathway. Taken together, this study suggests that match with a toxicant signature can assign a putative mechanism of action to the test compound if is established a database containing response patterns to various toxic compounds.

Progressive Iterative Forward and Backward (PIFAB) Search Method to Estimate Path-Travel Time on Freeways Using Toll Collection System Data (고속도로 경로통행시간 산출을 위한 전진반복 전후방탐색법(PIFAB)의 개발)

  • NamKoong, Seong
    • Journal of Korean Society of Transportation
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    • v.23 no.5 s.83
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    • pp.147-155
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    • 2005
  • The purpose of this paper is to develop a method for estimation of reliable path-travel time using data obtained from the toll collection system on freeways. The toll collection system records departure and arrival time stamps as well as the identification numbers of arrival and destination tollgates for all the individual vehicles traveling between tollgates on freeways. Two major issues reduce accuracy when estimating path-travel time between an origin and destination tollgate using transaction data collected by the toll collection system. First, travel time calculated by subtracting departure time from arrival time does not explain path-travel time from origin tollgate to destination tollgate when a variety of available paths exist between tollgates. Second, travel time may include extra time spent in service and/or rest areas. Moreover. ramp driving time is included because tollgates are installed before on-ramps and after off-ramps. This paper describes an algorithm that searches for arrival time when departure time is given between tollgates by a Progressive Iterative Forward and Backward (PIFAB) search method. The algorithm eventually produces actual path-travel times that exclude any time spent in service and/or rest areas as well as ramp driving time based on a link-based procedure.

Gene expression and SNP identification related to leaf angle traits using a genome-wide association study in rice (Oryza sativa L.) (GWAS 분석을 이용한 벼 지엽각 관련 SNP 동정 및 발현 분석)

  • Kim, Me-Sun;Yu, Yeisoo;Kang, Kwon-Kyoo;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.45 no.1
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    • pp.17-29
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    • 2018
  • This study was conducted to investigate a morphological trait in 294 rice accessions including Korean breeding lines. We also carried out a genome-wide association study (GWAS) to detect significant single nucleotide polymorphism markers and candidate genes affecting major agronomic traits. A Manhattan plot analysis of GWAS using morphological traits showed that phenotypic and statistical significance was associated with a chromosome in each group. The significance of SNPs that were detected in this study was investigated by comparing them with those found previously studied QTL regions related to agronomic traits. As a result, SNP (S8-19815442), which is significant with regard to leaf angle, was located in the known QTL regions. To observe gene mutations related to leaf angle in a candidate gene, Os08g31950, its sequences were compared with sequences in previously selected rice varieties. In Os08g31950, a single nucleotide mutation occurred in one region. To compare relative RNA expression levels of candidate gene Os08g31950, obtained from GWAS analysis of 294 rice accessions and related to lateral leaf angle, we investigated relative levels by selecting 10 erect leaf angle varieties and 10 horizontal leaf angle varieties and examining real-time PCR. In Os08g31950, a high level of expression and various expression patterns were observed in all tissues. Also, Os08g31950 showed higher expression levels in the erect leaf angle variety group and higher expression rates in the leaf than in the root. The candidate gene detected through GWAS would be useful in developing new rice varieties with improved yield potential through future molecular breeding.