• Title/Summary/Keyword: Fluorescence Response

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Feature Extraction for Protein Pattern Using Fuzzy Integral (퍼지적분을 이용한 단백질패턴에 관한 특징추출)

  • Song, Young-Jun;Kwon, Heak-Bong;Kim, Mi-Hye
    • The Journal of the Korea Contents Association
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    • v.7 no.1
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    • pp.40-47
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    • 2007
  • In the protein macro array image, it is important to find out the feature of the each protein chip. A decision error by the personal sense of sight occurred from long time observation while making an experiment in many protein chip image. So the feature extraction is needed by a simulator. In the case of feature analysis for macro array scan image the efficiency is maximized. In the fluorescence scan image, the response for each cell have been depend on R, G, B distribution of color image. But it is difficult to be classified as one color feature in the case of mixed color image. In this paper, the response color of a protein chip is classified according to the fuzzy integral value with respect to fuzzy measure as the user desired color. The result of the experiment for the macro array fluorescence image with the Scan Array 5000 shows that the proposed method using the fuzzy integral is important fact to be make decision for the ambiguous color.

Fluorometric Detection of Low-Abundance EGFR Exon 19 Deletion Mutation Using Tandem Gene Amplification

  • Kim, Dong-Min;Zhang, Shichen;Kim, Minhee;Kim, Dong-Eun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.5
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    • pp.662-667
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    • 2020
  • Epidermal growth factor receptor (EGFR) mutations are not only genetic markers for diagnosis but also biomarkers of clinical-response against tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). Among the EGFR mutations, the in-frame deletion mutation in EGFR exon 19 kinase domain (EGFR exon 19-del) is the most frequent mutation, accounting for about 45% of EGFR mutations in NSCLCs. Development of sensitive method for detecting the EGFR mutation is highly required to make a better screening for drug-response in the treatment of NSCLC patients. Here, we developed a fluorometric tandem gene amplification assay for sensitive detection of low-abundance EGFR exon 19-del mutant genomic DNA. The method consists of pre-amplification with PCR, thermal cycling of ligation by Taq ligase, and subsequent rolling circle amplification (RCA). PCR-amplified DNA from genomic DNA samples was used as splint DNA to conjugate both ends of linear padlock DNA, generating circular padlock DNA template for RCA. Long stretches of ssDNA harboring multiple copies of G-quadruplex structure was generated in RCA and detected by thioflavin T (ThT) fluorescence, which is specifically intercalated into the G-quadruplex, emitting strong fluorescence. Sensitivity of tandem gene amplification assay for detection of the EGFR exon 19-del from gDNA was as low as 3.6 pg, and mutant gDNA present in the pooled normal plasma was readily detected as low as 1% fraction. Hence, fluorometric detection of low-abundance EGFR exon 19 deletion mutation using tandem gene amplification may be applicable to clinical diagnosis of NSCLC patients with appropriate TKI treatment.

Evaluation through Photochemical Response Analysis on Growth Enhancing Effect of Decomposed Hatchery Waste Egg for Red Pepper (광화학적 반응 분석을 통한 부화장 폐달걀 분해 액비의 고추 생장촉진효과 평가)

  • Yoo, Sung Yung;Kang, Hong Gyu;Yoo, Jae Hong;Lee, Jeon Gyu;Shim, Myoung Yong
    • Korean Journal of Environmental Biology
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    • v.34 no.3
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    • pp.161-168
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    • 2016
  • In this study, growth enhancing effect of hatchery waste egg decomposed liquid fertilizer in pepper plant cultivation through chlorophyll fluorescence (O-J-I-P) analysis. In a whole growth period, egg decomposed fertilizer treated pepper grew well than non treated plant, though it was not statistically significantly different. Amount of chlorophyll fluorescence of non treated plant was higher thant that of fertilizer treated plant. It is determined that eventually lead to increased photosynthesis. In this study, six parameters, Fo, ABS/RC, RC/ABS, TRo/RC, DI0/RC, and DF Total ABS were the important factors represent efficiency of photochemical responses of pepper plant treated with hatchery waste egg decomposed fertilizer.

Microfluidic Fabrication of Conjugated Polymer Sensor Fibers (미세유동을 이용한 공액 고분자 센서 섬유 제작)

  • Yoo, Imsung;Song, Simon
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.38 no.10
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    • pp.853-858
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    • 2014
  • We propose a fabrication method for polydiacetylene (PDA)-embedded hydrogel microfibers on a microfluidic chip. These fibers can be applied to the detection of cyclodextrines (CDs), which are a family of sugar and aluminum ions. PDA, a family of conjugated polymers, has unique characteristics when used for a sensor, because it undergoes a blue-to-red color transition and nonfluorescence-to-fluorescence transition in response to environmental stimulation. PDAs have different sensing characteristics depending on the head group of PCDA. By taking advantage of ionic crosslinking-induced hydrogel formation and the 3D hydrodynamic focusing effect on a microfluidic chip, PCDA-EDEA-derived diacetylene (DA) monomer-embedded microfibers were successfully fabricated. UV irradiation of the fibers afforded blue-colored PDA, and the resulting blue PDA fibers underwent a phase transition to red and emitted red fluorescence upon exposure to CDs and aluminum ions. Their fluorescence intensity varied depending on the CDs and aluminum ion concentrations. This phase transition was also observed when the fibers were dried.

Computational and experimental characterization of estrogenic activities of 20(S, R)-protopanaxadiol and 20(S, R)-protopanaxatriol

  • Zhang, Tiehua;Zhong, Shuning;Hou, Ligang;Wang, Yongjun;Xing, XiaoJia;Guan, Tianzhu;Zhang, Jie;Li, Tiezhu
    • Journal of Ginseng Research
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    • v.44 no.5
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    • pp.690-696
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    • 2020
  • Background: As the main metabolites of ginsenosides, 20(S, R)-protopanaxadiol [PPD(S, R)] and 20(S, R)-protopanaxatriol [PPT(S, R)] are the structural basis response to a series of pharmacological effects of their parent components. Although the estrogenicity of several ginsenosides has been confirmed, however, the underlying mechanisms of their estrogenic effects are still largely unclear. In this work, PPD(S, R) and PPT(S, R) were assessed for their ability to bind and activate human estrogen receptor α (hERα) by a combination of in vitro and in silico analysis. Methods: The recombinant hERα ligand-binding domain (hERα-LBD) was expressed in E. coli strain. The direct binding interactions of ginsenosides with hERα-LBD and their ERα agonistic potency were investigated by fluorescence polarization and reporter gene assays, respectively. Then, molecular dynamics simulations were carried out to simulate the binding modes between ginsenosides and hERα-LBD to reveal the structural basis for their agonist activities toward receptor. Results: Fluorescence polarization assay revealed that PPD(S, R) and PPT(S, R) could bind to hERα-LBD with moderate affinities. In the dual luciferase reporter assay using transiently transfected MCF-7 cells, PPD(S, R) and PPT(S, R) acted as agonists of hERα. Molecular docking results showed that these ginsenosides adopted an agonist conformation in the flexible hydrophobic ligand-binding pocket. The stereostructure of C-20 hydroxyl group and the presence of C-6 hydroxyl group exerted significant influence on the hydrogen bond network and steric hindrance, respectively. Conclusion: This work may provide insight into the chemical and pharmacological screening of novel therapeutic agents from ginsenosides.

Highly Sensitive Fluorescent Probes for the Quantitative Determination of Singlet Oxygen (1O2)

  • Ahmed, Syed Rahin;Koh, Kwang-Nak;Kang, Nam-Lyong;Lee, Jae-Beom
    • Bulletin of the Korean Chemical Society
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    • v.33 no.5
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    • pp.1608-1612
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    • 2012
  • Singlet oxygen ($^1O_2$) is an important species for oxidation in biological processes. $^1O_2$ is implicated in the genotoxic effect, and plays an important role in the cell-signaling cascade and in the induction of gene expression. However, the rapid detection of $^1O_2$ in biological environments with sufficient specificity and sensitivity is hampered by its extremely low emission probability. Here, a layer-by-layer (LbL) film of CdTe quantum dots (QDs), polymers, and ascorbate have been designed as a rapid, highly selective, and sensitive fluorescence probe for $^1O_2$ detection. Upon reaction with $^1O_2$, the probe exhibits a strong photoluminescence (PL) response even at trace levels. This remarkable PL change should enable the probe to be used for $^1O_2$ detection in many chemical and biological systems and as an environmental sensor.

Microfilter Chip Fabrication for Bead-Based Immunoassay (비드를 이용한 면역분석용 마이크로필터 칩의 제작)

  • Lee, Seung-Woo;Ahn, Yoo-Min;Chai, Young-Gyu
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.28 no.9
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    • pp.1429-1434
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    • 2004
  • Immunoassay is one of the important analytical methods for clinical diagnoses and biochemical studies, but needs a long time, troublesome procedures and expensive reagents. In this study, therefore, we propose the micro filter chip with microbeads for immunoassay, which has pillar structures. The advantage of the proposed micro filter chip is to use simple fabrication process and cheap materials. The mold was made by the photolithography technique with Si wafer and negative photoresist SU-8. The replica was made of PDMS, bonded on the pyrex glass. The micro filter chip consists of inlet channel, filter chamber and outlet channel. HBV (Hepatitius B virus) monoclonal antibody (Ag1) labeled with biotin were immobilized onto streptavidin coated beads of 30∼50 $\mu$m size. Fluorescein isothiocyanate (FITC)-labeled HBV monoclonal antibody (Ag8) was used to detect HBsAg (Hebatitis B virus surface Antigen), and fluorescence intensity was monitored by epi-fluorescence microscope. In this study, the immune response of less than 30 min was obtained with with the use of 100 $m\ell$ of sample.

Construction and Expression of an Eukaryotic Expression Vector Containing the IER3 Gene

  • Wang, Zhen;Yu, Hong-Sheng;Yao, Ru-Yong;Qiu, Wen-Sheng;Yue, Lu;Sui, Ai-Hua;Liu, Xiang-Ping;Liu, Shi-Hai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.507-510
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    • 2013
  • Background: More and more research indicate that the immediately early response gene 3 (IER3) is involved inmany biological provesses, such as apoptosis and immunoreaction, as well as viral infection, tumorigenesis and tumour progression. Methods: Here we describe the construction of an eukaryotic expression vector containing IER3 gene and its expression in A549 cells as assessed through fluorescence microscopyand Western-blotting. Results: Fluorescence detection displayed that GFP in cytoplasm was high during 48 and 72 hours post-transfection. In addition, Western blotting showed significant increase in IER3 gene expression in the transfected cells compared with controls. Conclusion: The recombinate plasmid expression vector was constructed successfully, which may provide a basis for further exploration of function of IER3 in lung cancer.

A Cyan Fluorescent Protein Gene (cfp)-Transgenic Marine Medaka Oryzias dancena with Potential Ornamental Applications

  • Vu, Nguyen Thanh;Cho, Young Sun;Lee, Sang Yoon;Kim, Dong Soo;Nam, Yoon Kwon
    • Fisheries and Aquatic Sciences
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    • v.17 no.4
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    • pp.479-486
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    • 2014
  • To evaluate their potential utility as an ornamental organism, novel transgenic marine medaka Oryzias dancena strains with a highly vivid fluorescent phenotype were established through transgenesis of a cyan fluorescent protein gene (cfp) driven by the endogenous fast skeletal myosin light chain 2 gene (mlc2f) promoter. The transgenic marine medaka strains possessed multiple copies of transgene integrants and passed their fluorescent transgenes successfully to subsequent generations. Transgenic expression in skeletal muscles at both the mRNA and phenotypic levels was, overall, dependent upon transgene copy numbers. In the external phenotype, an authentic fluorescent color was dominant in the skeletal muscles of the transgenic fish and clearly visible to the unaided eye. The phenotypic fluorescent color presented differentially in response to different light-irradiation sources; the transgenics displayed a yellow-green color under normal daylight or white room light conditions, a strong green-glowing fluorescence under ultraviolet light, and a cyan-like fluorescence under blue light from a light-emitting diode.

Heat stress protection in Aspen sp1 transgenic Arabidopsis thaliana

  • Zhu, Bo;Xiong, Ai-Sheng;Peng, Ri-He;Xu, Jing;Zhou, Jun;Xu, Jin-Tao;Jin, Xiao-Fen;Zhang, Yang;Hou, Xi-Lin;Yao, Quan-Hong
    • BMB Reports
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    • v.41 no.5
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    • pp.382-387
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    • 2008
  • It is known that the stable protein 1 (SP1) detected in aspen plants remains soluble upon boiling and that sp1 expression in transgenic aspen is resistant to salt stress. Presently, we analyzed the effect of expression of SP1 in Arabidopsis thaliana plants and their response to high temperature stress. After $45^{\circ}C$ for 16 h, relative to wild type plants, sp1 transgenic plants exhibited stronger growth and were better in several physiological properties including chlorophyII, chlorophyII fluorescence, water content, proline content, and malondialdehyde content. These preliminarily results suggest that the over-expression of SP1 may notably enhance heat-tolerant level of transgenic A. thaliana plants.