• Title/Summary/Keyword: m/RT

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Development of a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay for the Simultaneous Detection of Three Viruses in Leguminous Plants

  • Park, Chung Youl;Min, Hyun-Geun;Lee, Hong-Kyu;Maharjan, Rameswor;Yoon, Youngnam;Lee, Su-Heon
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.348-352
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    • 2018
  • A multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay was developed for the detection of Clover yellow vein virus (ClYVV), Peanut mottle virus (PeMoV), and Tomato spotted wilt virus (TSWV), which were recently reported to infect soybean and azuki bean in Korea. Species-specific primer sets were designed for the detection of each virus, and their specificity and sensitivity were tested using mixed primer sets. From among the designed primer sets, two combinations were selected and further evaluated to estimate the detection limits of uniplex, duplex, and multiplex RT-PCR. The multiplex RT-PCR assay could be a useful tool for the field survey of plant viruses and the rapid detection of ClYVV, PeMoV, and TSWV in leguminous plants.

Time-triggered Message-triggered Object Programming Scheme and Its Support Middleware

  • (Kane)Kim, K.H.;Kim, M.H.;Rim, K.W.
    • Korea Information Processing Society Review
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    • v.11 no.6
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    • pp.8-24
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    • 2004
  • OO RT programming is a technology expected to flourish in this quarter of the 21st century. Currently, its youthfulness is indicated by the insufficient availability of the support middleware and the associated API, let alone language compilers. The middleware providing fault-tolerant execution support is in its infancy. The advances in OO RT distributed programming will also enable large-scale RT simulations. The research community dealing with this technology area is expected to grow continuously for foreseeable future and consequent accelerations of the technology advances will in turn accelerate the development of many new types of sophisticated RT DC applications as well as realization of advanced types of ubiquitous computing societies.

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Use of RT-PCR for CK20 and CEA mRNA to Detect of Micrometastasis in the Draining Venous Blood and the Peritoneum in Gastric Cancer Patients (위암에서 CK20과 CEA의 RT-PCR을 이용한 수술조작 전후 유출 정맥과 복막에서의 미세전이의 검출)

  • Joo Jai Kyun;Lee Ji Hee;Koh Yang Seok;Kim Jung Chul;Ryu Seong Yeob;Kim Dong Yi;Kim Young Jin
    • Journal of Gastric Cancer
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    • v.3 no.3
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    • pp.128-133
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    • 2003
  • Purpose: The benefits of the 'no-touch' isolation techniquethat is usually performed to prevent the circulation of tumor cells are not evident. The aim of this study was to determine whether the no-touch isolation technique for treating gastrointestinal cancers could prevent the circulation of tumor cells detected by reverse transcriptase polymerase chain reaction (RT-PCR). Matrials and Methods: By using RT-PCR to amplify mRNAs for two specific epithelial markers, carcinoembryonic antigen (CEA) and cytokeratin 20 (CK-20), we examined 34 gastric cancer patients who had been histologically diagnosed and 22 patients had undergone serosal and peritoneal brushing. Results: In 10 ($29.4\%$) of the 34 gastric cancer patients, we detected CK20 mRNA before manipulation, and in 17 ($51.5\%$) of those patients, after we detected it. The density of the CK20 mRNA band was increased in 11 cases ($33.3\%$) and the density was decreased in 2 cases ($6.1\%$). In 16 ($48.5\%$) of the 34 gastric cancer patients, we detected CEA mRNA before manipulation, and in 17 ($51.5\%$) patients after we detected it. The density of the CEA mRNA band was increased in 8 cases ($24.2\%$) and decreased in 3 cases ($9.1\%$). Conclusion: These result suggest that the ' no-touch isolation technique ' might be useful when operating on advanced gastric cancer patients and that serosal or Douglas pouch brushing can be used to determine the status of micrometastasis.

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Validation of Stem-loop RT-qPCR Method on the Pharmacokinetic Analysis of siRNA Therapeutics (Stem-loop RT-qPCR 분석법을 이용한 siRNA 치료제의 생체시료 분석법 검증 및 약물 동태학적 분석)

  • Kim, Hye Jeong;Kim, Taek Min;Kim, Hong Joong;Jung, Hun Soon;Lee, Seung Ho
    • Journal of Life Science
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    • v.29 no.6
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    • pp.653-661
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    • 2019
  • The first small interfering RNA (siRNA) therapeutics have recently been approved by the Food and Drug Administration in the U.S., and the demand for a new RNA therapeutics bioanalysis method-which is essential for pharmacokinetics, including the absorption, distribution, metabolism, and excretion of siRNA therapeutics-is rapidly increasing. The stem-loop real-time qPCR (RT-qPCR) assay is a useful molecular technique for the identification and quantification of small RNA (e.g., micro RNA and siRNA) and can be applied for the bioanalysis of siRNA therapeutics. When the anti-HPV E6/E7 siRNA therapeutic was used in preclinical trials, the established stem-loop RT-qPCR assay was validated. The limit of detection was sensitive up to 10 fM and the lower limit of quantification up to 100 fM. In fact, the reliability of the established test method was further validated in three intra assays. Here, the correlation coefficient of $R^2$>0.99, the slope of -3.10 ~ -3.40, and the recovery rate within ${\pm}20%$ of the siRNA standard curve confirm its excellent robustness. Finally, the circulation profiles of siRNAs were demonstrated in rat serum, and the pharmacokinetic properties of the anti-HPV E6/E7 siRNA therapeutic were characterized using a stem-loop RT-qPCR assay. Therefore, the stemloop RT-qPCR assay enables accurate, precise, and sensitive siRNA duplex quantification and is suitable for the quantification of small RNA therapeutics using small volumes of biological samples.

Interleukin-8 gene expression in the human colon epithelial cell line, HT-29, exposed to Entamoeba histolytica (이질아메바에 의한 인체 대장상피세포주 HT-29에서의 interleukin-8 유전자의 발현)

  • 김정목;정현채
    • Parasites, Hosts and Diseases
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    • v.33 no.4
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    • pp.357-364
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    • 1995
  • The protozoan parasite, Entcmoeba histoIWticc, is one of major causative agents of intestinal disease all over the world. In acute experimental infection, the early host response to 5. histoIHtica is characterized by an infiltration of neutrophils. However, the chemotactic signal for this response is not well known. Based on the (jading that human epithelial cells produce the potent neutrophil chemoattractant and activator, interleukin-8 (IL-8), IL-8 gene expression was examined thoroughly in human colon epithelial cells exposed to 5. histolvtica trophozoites. Cellular RNAs were extracted from HT-29 or Caco-2 human colon epithelial cells exposed to 5. histoLvtica trophozoites for 30 minutes, 1 and 3 hours. IL-8 mRNA transcripts were measured by reverse transcriptional polprnerase chain reaction (RT-PCR) using synthetic standard RNA. The number of IL-8 mRNA molecules increased from 30 minutes to 3 hours of exposure period, reaching 3.1 H 107 molecules/ug of total RNA. Expression pattern of IL-8 mRNA transcripts was parallel to the amounts of IL-8 protein measured by enzyme-linked immunosorbent assay (ELISA) . Lysates of 5. histoIVtica also induced expression of mRNA for IL-8 in colon epithelial cells. These results sugf:esc that acute inflammatory reaction by 5. histoIVticc may be initially triggered by proinflammatory cytokines such as IL-8 secreted from epithelial cells of the colon.

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Responses of Photosynthetic Efficiency and Ascorbate Peroxidase Induced by Salt Stress in Rice (Oryza sativa L.) (벼의 salt stress에 의해 유도된 산화 stress에 대한 ascorbate peroxidase 반응)

  • Koo, Jeung-Suk;Im, Kyoung-Nam;Chun, Hyun-Sik;Lee, Chin-Bum
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1173-1180
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    • 2010
  • We investigated changes in photosynthesis and activity of ascorbate peroxidase (APX) that scavenges ROS as responses to oxidative stress induced by salinity in rice (Oryza sativa L.). Photosynthetic efficiency of rice leaves, monitored in terms of Fv/Fm, declined with the increase of salt concentration (100-300 mM NaCl). Salinity caused an increase of $H_2O_2$ in leaves of rice, with an increase of APX activity. Among total APX isoforms, an isoform of stromal-APX 1 in leaves of rice was completely inactivated by 300 mM NaCl, but was not affected by chilling or drought. The results suggest that salt stress acts in quite a different mechanism in relation to the activity of stromal-APX from that of other stresses such chilling and drought. We carried out RT-PCR for analysis of genes expression of APX isoforms as affected by salt stress. The expression of cytosolic APX/thylakoid-bound APX genes in leaves of rice exposed to salt stress was increased, while stromal APX gene expression rapidly declined.

Expression of Vitellogenin Gene by Androgens in Rasinbow Trout, Oncorhynchus mykiss (웅성호르몬에 의한 무지개송어의 vitellogenin 유전자 발현)

  • 권혁추;윤종만;이종영
    • Journal of Aquaculture
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    • v.13 no.1
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    • pp.79-85
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    • 2000
  • The effects of estrogen and androgens on Vg gene expression were examined in primary hepatocyte culture and livers of the immature male trout. Specific primers of Vg cDNA were designed with already reported Vg gene nucleotide sequences. PCR product was sequenced and verified with Vg cDNA of rainbow trout. Total RNA was extracted from the cultured hepatocytes and livers of steroid-treated rainbow trout and then it was analyzed by reverse transcriptase- polymerase chain reaction (RT-PCR) analysis. The Vg mRNA and Vg protein synthesis were increased in rainbow trout in vivo and in vitro with E$_2$ and methyltestosterone (MT) There were dose and time-related effects of E$_2$ and MT on vitellogesis. Androgens such as progesterone androsterone and testosterone also stimulated Vg mRNA expression in vitro. The results show that androgens as well as E$_2$ can induce expression of Vg mRNA in trout in vivo and in vitro.

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Effects of Diffusion Layer (DL) and ORR Catalyst (MORR) on the Performance of MORR/IrO2/DL Electrodes for PEM-Type Unitized Regenerative Fuel Cells

  • Choe, Seunghoe;Lee, Byung-Seok;Jang, Jong Hyun
    • Journal of Electrochemical Science and Technology
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    • v.8 no.1
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    • pp.7-14
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    • 2017
  • This study aims to examine the influences of substrates/diffusion layers (DL) and oxygen reduction reaction catalysts ($M_{ORR}$) on the performance of $M_{ORR}/IrO_2$/DL-type bifunctional oxygen electrodes for use in polymer electrolyte membrane (PEM)-type unitized regenerative fuel cells (URFC). The $M_{ORR}/IrO_2$/DL electrodes were prepared via two sequential steps: anodic electrodeposition of $IrO_2$ on various DLs and fabrication of $M_{ORR}$ layers (Pt, Pd, and Pt-Ru) by spraying on $IrO_2/DL$. Experiments using different DLs, with Pt as the $M_{ORR}$, revealed that the roughness factor of the DL mainly determined the electrode performance for both water electrolyzer (WE) and fuel cell (FC) operations, while the contributions of porosity and substrate material were insignificant. When Pt-Ru was utilized as the $M_{ORR}$ instead of Pt, WE performance was enhanced and the electrode performance was assessed by analyzing round-trip efficiencies (${\varepsilon}_{RT}$) at current densities of 0.2 and $0.4A/cm^2$. As a result, using Pt-Ru instead of Pt alone provided better ${\varepsilon}_{RT}$ at both current densities, while Pd resulted in very low ${\varepsilon}_{RT}$. Improved efficiency was related to the additional catalytic action by Ru toward ORR during WE operation.

Thermal Interference Modeling in the Aquifer Thermal Energy Storage Systems (대수층 축열 시스템의 열 간섭 모델링)

  • Kim, Jong-chan;Lee, Young-min;Yoon, Woon-Sang;Koo, Min-Ho;Keehm, Young-seuk
    • 한국신재생에너지학회:학술대회논문집
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    • 2009.06a
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    • pp.596-599
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    • 2009
  • 대수층 축열 시스템(ATES)의 효율은 지중에 저장된 열 사이의 간섭에 의해 결정이 되며, 열 간섭에 영향을 미치는 요인들은 시추공 간 이격거리, 대수층의 수리 전도도, 주입/양수량 등이 있다. 따라서 이 연구에서는 지하수-지열 거동 모델링을 통하여 열 간섭이 일어나는 현상과 열 간섭이 시스템 효율에 미치는 영향을 분석하였다. 모델링 결과로부터 이격거리는 작을수록 그리고 대수층의 수리전도도와 주입/양수량은 클수록 열 간섭이 잘 일어나는 것을 확인하였고, 열 간섭 계수가 1일 때는 시스템 효율이 상대적으로 크게 낮아지고, 열 간섭 계수가 1 미만일 때는 변화가 미미한 것을 확인하였다. ATES 를 이용한 냉난방 시스템을 시공 하고 있는 안성 연구지역에 대한 장기 예측 모델링을 수행 하였다. 모델링 결과 이격거리가 80 m 이고, 주입/양수량이 100 $m^3/day$ 일 때, 시스템 가동 7년경과 뒤 여름철과 겨울철에 계산된 시스템 효율은 각각 36 RT 와 31 RT 로 나타났다. 따라서 주입/양수량을 100 $m^3/day$로 했을 때, 냉난방 대상건물의 필요 부하인 20 RT 를 충분히 충족할 수 있을 것으로 판단된다.

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Development of a novel reverse transcription PCR and its application to field sample testing for feline calicivirus prevalence in healthy stray cats in Korea

  • Kim, Sung Jae;Park, Yong Ho;Park, Kun Taek
    • Journal of Veterinary Science
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    • v.21 no.5
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    • pp.71.1-71.10
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    • 2020
  • Background: Feline calicivirus (FCV) is a major and highly infectious pathogen in cats worldwide. However, there have been limited studies about the status of FCV infections in Korea. Objectives: To investigate the current status of FCV infections in stray cats in Korea. Methods: A novel reverse transcription polymerase chain reaction (RT-PCR) assay was developed based on the conserved nucleotide sequences of reported FCV strains. Field swab samples were collected from 122 cats (2 hospital admitted cats and 120 stray cats) in 2016 and 2017. All the samples were tested by virus isolation and 2 different RT-PCRs, including the novel RT-PCR, for the detection of FCV. Results: The novel RT-PCR assay showed no cross-reactivity to the nucleic acids of the other feline pathogens tested, and the limit of detection was calculated as 100 TCID50/mL based on an in vitro assessment. The novel RT-PCR assay detected 5 positive samples from the 122 field samples, which showed perfect agreement with the results of the virus isolation method. In contrast, another RT-PCR assay used in a previous study in Korea detected no positive samples. The prevalence of FCV infection in stray cats was 2.5% (3/120) based on the results of virus isolation and the novel RT-PCR assays. Conclusions: The current study is the first report of the detection and prevalence of FCV in stray cats in Korea. The novel RT-PCR assay developed in this study showed high sensitivity and specificity, which indicates a useful diagnostic assay to identify FCV infection in cats.