• Title/Summary/Keyword: sprA

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Ginsenoside Rd protects cerebral endothelial cells from oxygen-glucose deprivation/reoxygenation induced pyroptosis via inhibiting SLC5A1 mediated sodium influx

  • Li, Suping;Yu, Nengwei;Xu, Fei;Yu, Liang;Yu, Qian;Fu, Jing
    • Journal of Ginseng Research
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    • v.46 no.5
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    • pp.700-709
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    • 2022
  • Background: Ginsenoside Rd is a natural compound with promising neuroprotective effects. However, the underlying mechanisms are still not well-understood. In this study, we explored whether ginsenoside Rd exerts protective effects on cerebral endothelial cells after oxygen-glucose deprivation/reoxygenation (OGD/R) treatment and its potential docking proteins related to the underlying regulations. Method: Commercially available primary human brain microvessel endothelial cells (HBMECs) were used for in vitro OGD/R studies. Cell viability, pyroptosis-associated protein expression and tight junction protein degradation were evaluated. Molecular docking proteins were predicted. Subsequent surface plasmon resonance (SPR) technology was utilized for validation. Flow cytometry was performed to quantify caspase-1 positive and PI positive (caspase-1+/PI+) pyroptotic cells. Results: Ginsenoside Rd treatment attenuated OGD/R-induced damage of blood-brain barrier (BBB) integrity in vitro. It suppressed NLRP3 inflammasome activation (increased expression of NLRP3, cleaved caspase-1, IL-1β and GSDMD-N terminal (NT)) and subsequent cellular pyroptosis (caspase-1+/PI + cells). Ginsenoside Rd interacted with SLC5A1 with a high affinity and reduced OGD/R-induced sodium influx and potassium efflux in HBMECs. Inhibiting SLC5A1 using phlorizin suppressed OGD/R-activated NLRP3 inflammasome and pyroptosis in HBMECs. Conclusion: Ginsenoside Rd protects HBMECs from OGD/R-induced injury partially via binding to SLC5A1, reducing OGD/R-induced sodium influx and potassium efflux, thereby alleviating NLRP3 inflammasome activation and pyroptosis.

Microcontact Printing of Biotin for Selective Immobilization of Streptavidin-fused Proteins and SPR Analysis

  • Lee, Sang-Yup;Park, Jong-Pil;Lee, Seok-Jae;Park, Tae-Jung;Lee, Kyung-Bok;Park, Insung S.;Kim, Min-Gon;Chung, Bong-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.2
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    • pp.137-142
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    • 2004
  • In this study, a simple procedure is described for patterning biotin on a glass substrate and then selectively immobilizing proteins of interest onto the biotin-patterned surface. Microcontact printing (CP) was used to generate the micropattern of biotin and to demonstrate the selective immobilization of proteins by using enhanced green fluorescent protein (EGFP) as a model protein, of which the C-terminus was fused to a core streptavidin (cSA) gene of Streptomyces avidinii. Confocal fluorescence microscopy was used to visualize the pattern of the immobilized protein (EGFP-cSA), and surface plasmon resonance was used to characterize biological activity of the immobilized EGFP-cSA. The results suggest that this strategy, which consists of a combination of $\mu$CP and cSA-fused proteins. is an effective way for fabricating biologically active substrates that are suitable for a wide variety of applications. one such being the use in protein-protein assays.

One-step phyto-mediated fabrication of silver nanoparticles and its anti-microbial properties

  • Velmurugan Palanivel;Sung-Chul Hong;Veera Ravi Arumugam;Sivakumar Subpiramaniyam;Pyong-In Yi;Seong-Ho Jang;Jeong-Min Suh;Eun-Sang Jung;Je-Sung Park
    • Advances in nano research
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    • v.14 no.4
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    • pp.391-397
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    • 2023
  • This manuscript describes the one-step eco-friendly green fabrication of silver nanoparticles (AgNPs) through the in-situ bio-reduction of an aqueous solution of silver nitrate using Syzygium aromaticum leaf extract. UV-vis spectroscopy shows a characteristic SPR peak around 442 nm. FTIR spectroscopy showed that the AgNPs were capped with bioactive phyto-molecules. TEM images revealed oval and spherical particles with a mean diameter of ~12.6 nm. XRD analysis revealed crystalline and face-cantered cubic AgNPs. The phytosynthesized AgNPs showed broad-spectrum anti-microbial activity against two foodborne pathogenic bacteria, Listeria monocytogenes and Staphylococcus aureus. The AgNPs showed a prominent ability to inhibit biofilms formed by L. monocytogenes and S. aureus in laboratory conditions through a crystal violet assay. The results suggest that the AgNPs could be a novel nanotool to develop effective antimicrobial and anti-biofilm agents in food preservation.

A Review of SERS for Biomaterials Analysis Using Metal Nanoparticles (바이오 물질 분석을 위한 금속 나노입자를 이용한 SERS 분석 연구동향)

  • Jang, Eue-Soon
    • Ceramist
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    • v.22 no.3
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    • pp.281-300
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    • 2019
  • Surface enhanced Raman scattering (SERS) was first discovered in 1974 by an unexpected Raman signal increase from Pyridine adsorbed on rough Ag electrode surfaces by the M. Fleishmann group. M. Moskovits group suggested that this phenomenon could be caused by surface plasmon resonance (SPR), which is a collective oscillation of free electrons at the surface of metal nanostructures by an external light source. After about 40 years, the SERS study has attracted great attention as a biomolecule analysis technology, and more than 2500 new papers and 500 review papers related to SERS topic have been published each year in recently. The advantages of biomaterials analysis using SERS are as follows; ① Molecular level analysis is possible based on unique fingerprint information of biomolecule, ② There is no photo-bleaching effect of the Raman reporters, allowing long-term monitoring of biomaterials compared to fluorescence microscopy, ③ SERS peak bandwidth is approximately 10 to 100 times narrower than fluorescence emission from organic phosphor or quantum dot, resulting in higher analysis accuracy, ④ Single excitation wavelength allows analysis of various biomaterials, ⑤ By utilizing near-infrared (NIR) SERS-activated nanostructures and NIR excitation lasers, auto-fluorescence noise in the visible wavelength range can be avoided from in vivo experiment and light damage in living cells can be minimized compared to visible lasers, ⑥ The weak Raman signal of the water molecule makes it easy to analyze biomaterials in aqueous solutions. For this reason, SERS is attracting attention as a next-generation non-invasive medical diagnostic device as well as substance analysis. In this review, the principles of SERS and various biomaterial analysis principles using SERS analysis will be introduced through recent research papers.

Imaging of self-assembled monolayers by surface plasmon microscope (표면 플라즈몬 현미경을 이용한 자기조립 단분자막의 이미징)

  • 표현봉;신용범;윤현철;양해식;김윤태
    • Korean Journal of Optics and Photonics
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    • v.14 no.1
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    • pp.97-102
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    • 2003
  • Multi-channel images of 11-MUA(11-Mercaptoundecanoic acid) and 11-MUOH(11-Mercaptoundecanol) self-assembled monolayers were obtained by using two-dimensional surface plasmon resonance (SPR) absorption. The patterning process was simplified by exploiting direct photo-oxidation of thiol bonding (photolysis) instead of conventional photolithography. Sharper images were resolved by using a white light source in combination with a narrow bandpass filter in the visible region, minimizing the diffraction patterns on the images. The line profile calibration of the image contrast caused by different resonance conditions at each point on the sensor surface (at a fixed incident angle) enables us to discriminate the monolayer thickness in nanometer scale. Furthermore, there is no signal degradation such as photo bleaching or quenching, which are common in the detection methods based on fluorescence.

Electrochemical Detection of $17{\beta}-estradiol$ by using DNA Aptamer Immobilized Nanowell Gold Electrodes

  • Kim, Yeon-Seok;Jung, Ho-Sup;Lee, Hea-Yeon;Kawai, Tomoji;Gu, Man-Bock
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.88-92
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    • 2005
  • Aptamer is the single-stranded oligonucleotide which binds to various target molecules such as proteins, peptides, lipids and small organic molecules with high affinity and specificity. DNA aptamers specific for the $17{\beta}-estradiol$ were selected by SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random DNA library. These DNA aptamers have a high affinity to $17{\beta}-estradiol$ as an endocrine disrupting chemical. Nanowell and $200{\mu}m$ gold electrode were used as substrate for DNA aptamer immobilization and electrochemical analysis. Especially, nanowell gold electrode was fabricated by e-beam lithography. The size of single nanowell is 130nm and 40,000 nanowells were deposited on one gold electrode. The immobilization method was based on the interaction between the biotinylated aptamer and streptavidin deposited on gold electrode previously. Immobilization procedure was optimized by surface plasma resonance (SPR) and electrochemical analysis. After the immobilization of DNA aptamer on streptavidin modified gold electrode, $17{\beta}-estradiol$ solution was treated on aptamer immobilized gold electrode. The current of gold electrode was decreased by the binding of $17{\beta}-estradiol$ to DNA aptamer immobilized on gold electrode. However, in negative control experiments of 1-aminoanthraquinone and 2-methoxynaphthalene, the current was rarely decreased. And more sensitive data was obtained from nanowell gold electrode comparing with $200{\mu}m$ gold electrode.

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A Novel Photonic Crystal Fiber Sensor with Three D-shaped Holes Based on Surface Plasmon Resonance

  • Bing, Pibin;Sui, Jialei;Huang, Shichao;Guo, Xinyue;Li, Zhongyang;Tan, Lian;Yao, Jianquan
    • Current Optics and Photonics
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    • v.3 no.6
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    • pp.541-547
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    • 2019
  • A novel photonic crystal fiber (PCF) sensor with three D-shaped holes based on surface plasmon resonance (SPR) is analyzed in this paper. Three D-shaped holes are filled with the analyte, and the gold film is deposited on the side of three planes. The design of D-shaped holes with outward expansion can effectively solve the uniformity problem of metallized nano-coating, it is beneficial to the filling of the analyte and is convenient for real-time measurement of the analyte. Compared with the hexagonal lattice structure, the triangular arrangement of the clad air holes can significantly reduce the transmission loss of light and improve the sensitivity of the sensor. The influences of the air hole diameter, the distance between D-shaped holes and core, and the counterclockwise rotation angle of D-shaped holes on sensing performance are studied. The simulation results show that the wavelength sensitivity of the designed sensor can be as high as 10100 nm/RIU and the resolution can reach 9.9 × 10-6 RIU.

Sensing of the Insecticide Carbofuran Residues by Surface Plasmon Resonance and Immunoassay (표면플라즈몬공명과 효소면역분석법을 이용한 살충제 카보후란 잔류물 검출)

  • Yang G. M.;Cho N. H.
    • Journal of Biosystems Engineering
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    • v.30 no.6 s.113
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    • pp.333-339
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    • 2005
  • The pesticide is raising public interest in the world, because it causes damage to an environmental pollution and the human health remaining agricultural products and an ecosystem, in spite of the advantages. Particularly, each country restricts the residual pesticide and induces observance about the safety and usage standard so that they can control the amount of pesticide used and defend the safety of agricultural products. The habitual practice for the analysis of the residual pesticide depends on GC (gas chromatography), HPLC (high performance liquid chromatography) and GC/MS (gas chromatography/mass spectroscopy), which triturate the fixed quantity of samples, abstract and purify as a suitable organic solvent. These methods have the highly efficient in aspects of sensitivity and accuracy. On the other hand, they need the high cost, time consuming, much effort, expensive equipment and the skillful management. Carbofuran is highly toxic by inhalation and ingestion and moderately toxic by dermal absorption. As with other carbamate compounds, it is metabolized in the liver and eventually excreted in the urine. The half-life of carbofuran on crops is about 4 days when applied to roots, and longer than 4 days if applied to the leaves. This research was conducted to develop immunoassay for detecting carbofuran residue quickly on the basis of surface plasmon resonance and to evaluate the measurement sensitivity. Gold chip used was CM5 spreaded dextran on the surface. An applied antibody to Immunoassay was GST (glutathione-s-transferase). The association and the dissociation time were 176 second and 215 second between GST and carbofuran. The total analysis time using surface plasmon resonance was 13 minutes including regeneration time, on the other hand HPLC and GC/MS was 2 hours usually. The minimum detection limit of a permissible amount for carbofuran in the country is 0.1 ppm. The immunoassay method using surface plasmon resonance was 0.002 ppm.

A Study on the Pathological Syndromes and the Insights of the Patients in the Community Mental Health Centers (지역사회 정신보건시설 내 환자의 정신병리 증상과 병식에 관한 연구)

  • An, Hyo-Ja;Kim, Hyeon-Mi
    • The Korean Journal of Rehabilitation Nursing
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    • v.9 no.2
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    • pp.109-116
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    • 2006
  • Purpose: This study is a descriptive research in which the pathological syndromes of the schizophrenic patients in the community mental health centers, the degrees of the insights, and the correlations between them were examined. Method: The subjects included 114 SPR pt. people who were enrolled in community mental health centers located in D City. The research was conducted during the period from Aug. 1st, 2002 to Jul. 30th, 2004. The instruments were PANSS developed by Kay(1987) and SUMD developed by Amador(1993). Results: Among the pathological syndromes, positives ones were $12.64{\pm}3.80$ on the average, negative ones were $32.00{\pm}5.36$, and general pathological syndromes were $30.50{\pm}6.24$. In the evaluation of the insights, the mean score was $11.81{\pm}5.09$. There were some significant correlations between positive syndromes and general pathological ones (r=.572, p=.000), and negative syndromes and general pathological ones(r=.262, p=.029), while there was no significant correlation between the insights and the sub-measures of general pathological syndromes. Conclusion: Therefore, the schizophrenic patients taking advantage of the community mental health centers have more negative syndromes than those in hospitalized, and rehabilitation programs are needed to help them continuously. And the further study of the correlations between the pathological syndromes and the degrees of the insights are required, and still, it should be analyzed what effects the insight acquirement brings about to the improvements of the pathological syndromes after the application of the insight-oriented programs.

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Medium Optimization and Application of Affinity Column Chromatography for Trypsin Production from Recombinant Streptomyces griseus

  • Chi, Won-Jae;Song, Ju-Hyun;Oh, Eun-A.;Park, Seong-Whan;Chang, Yong-Keun;Kim, Eung-Soo;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1191-1196
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    • 2009
  • The production of Streptomyces griseus trypsin (SGT) by S. griseus IFO13350 transformed with the expression vector pWHM3-TR1R2, containing sprT encoding SGT and the two positive regulatory genes sgtR1 and sgtR2, was investigated in various media. Cultivation in Ferm-0 gave 1.4 times more trypsin activity than in C5/L medium. In addition, replacement of 2% glucose and 1% skim milk in Ferm-0 with 2% dextrin and 1% tryptone (designated Ferm-II) enhanced trypsin activity 4.1-fold. To simplify the purification process, the supernatant from the S. griseus transformant cultured in Ferm-II medium was fractionated with ammonium sulfate (25-55%), then subjected to Hitrap Benzamidine FF affinity column chromatography. The specific activity of SGT purified by one-step chromatography was 69,550 unit/mg protein and the overall purification yield was above 8%, indicating that this method is more effective than those previously reported. Purified SGT was most active at pH 8.0 and $50^{\circ}C$, and it maintained activity between pH 7.0 and 9.0 and at temperatures up to $70^{\circ}C$. These enzymatic properties are very similar to those of authentic eukaryotic trypsin purified from bovine pancreas.