• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.2079-2094
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• 2018

Sunflower trypsin inhibitor (SFTI) is a 14-amino-acid bicyclic peptide that contains a single internal disulfide bond. We initially constructed chimeras of SFTI with N-terminal secretion signals from the Escherichia coli OmpA and Pseudomonas aeruginosa ToxA, but only detected small amounts of protease inhibition resulting from these constructs. A substantially higher degree of protease inhibition was detected from a C-terminal SFTI fusion with E. coli YebF, which radiated more than a centimeter from an individual colony of E. coli using a culture-based inhibitor assay. Inhibitory activity was further improved in YebF-SFTI fusions by the addition of a trypsin cleavage signal immediately upstream of SFTI, and resulted in production of a 14-amino-acid, disulfide-bonded SFTI free in the culture supernatant. To assess the potential of the secreted SFTI to protect the ability of a cytotoxic protein to kill tumor cells, we utilized a tumor-selective form of the Pseudomonas ToxA (OTG-PE38K) alone and expressed as a polycistronic construct with YebF-SFTI in the tumor-targeted Salmonella VNP20009. When we assessed the ability of toxin-containing culture supernatants to kill MDA-MB-468 breast cancer cells, the untreated OTG-PE38K was able to eliminate all detectable tumor cells, while pretreatment with trypsin resulted in the complete loss of anticancer cytotoxicity. However, when OTG-PE38K was co-expressed with YebF-SFTI, cytotoxicity was completely retained in the presence of trypsin. These data demonstrate SFTI chimeras are secreted in a functional form and that co-expression of protease inhibitors with therapeutic proteins by tumor-targeted bacteria has the potential to enhance the activity of therapeutic proteins by suppressing their degradation within a proteolytic environment.

• Journal of The Korean Astronomical Society
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• v.50 no.5
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• pp.139-149
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• 2017

The Korea Astronomy and Space Science Institute plans to develop a coronagraph in collaboration with National Aeronautics and Space Administration (NASA) and to install it on the International Space Station (ISS). The coronagraph is an externally occulted one-stage coronagraph with a field of view from 3 to 15 solar radii. The observation wavelength is approximately 400 nm, where strong Fraunhofer absorption lines from the photosphere experience thermal broadening and Doppler shift through scattering by coronal electrons. Photometric filter observations around this band enable the estimation of 2D electron temperature and electron velocity distribution in the corona. Together with a high time cadence (<12 min) of corona images used to determine the geometric and kinematic parameters of coronal mass ejections, the coronagraph will yield the spatial distribution of electron density by measuring the polarized brightness. For the purpose of technical demonstration, we intend to observe the total solar eclipse in August 2017 with the filter system and to perform a stratospheric balloon experiment in 2019 with the engineering model of the coronagraph. The coronagraph is planned to be installed on the ISS in 2021 for addressing a number of questions (e.g., coronal heating and solar wind acceleration) that are both fundamental and practically important in the physics of the solar corona and of the heliosphere.

• The Bulletin of The Korean Astronomical Society
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• v.46 no.2
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• pp.45.1-45.1
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• 2021

Horizon Run 5 (HR5) is a cosmological hydrodynamical simulation which captures the properties of the Universe on aGpc scale while achieving a resolution of 1kpc. This enormous dynamic range allows us to simultaneously capture the physics of the cosmic web on very large scales and account for the formation and evolution of dwarf galaxies on much smaller scales. On the back of a remarkable achievement of this, we have finished to run follow-up simulations which have 2 times larger volume than before and are expected to complementary to some limitations of previous HR simulations both for the study on the large scale features and the expansion history in a distant Universe. For these simulations, we consider the sub-grid physics of radiative heating/cooling, reionization, star formation, SN/AGN feedbacks, chemical evolution and the growth of super-massive blackholes. In order to do this project, we implemented a hybrid MPI-OpenMP version of the RAMSES code, 'RAMSES-OMP', which is specifically designed for modern many-core many thread parallel systems. These simulation successfully reproduce various observation result and provide a large amount of statistical samples of Lyman-alpha emitters and protoclusters which are important to understand the formation and expansion history of early universe. These are invaluable assets for the interpretation of current ΛCDM cosmology and current/upcoming deep surveys of the Universe, such as the world largest narrow band imaging survey, ODIN (One-hundred-square-degree Dark energy camera Imaging in Narrow band).

• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1243-1250
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• 2010

A cDNA encoding a cysteine protease inhibitor (CPI) was isolated from the cDNA library of clamworm Perinereis aibuhitensis Grube. The deduced amino acid sequence analysis showed that the protein had 51%, 48%, and 48% identity with Zgc:153129 from Danio rerio, cystatin B from Theromyzon tessulatum, and the ChainA, stefin B tetramer from Homo sapiens, respectively. The gene was cloned into the intracellular expression vector pET-15b and expressed in Escherichia coli. The recombinant CPI (PA-CPI) was purified by affinity chromatography on Ni-charged resin and ion-exchange chromatography on DEAE-Sepharose FF. The relative molecular mass of PA-CPI was 16 kDa as deduced by SDS-PAGE. Activity analysis showed that the recombinant protein could inhibit the proteolytic activity of papain. A constitutive and secretive expression vector was also constructed, and the cDNA encoding CPI was subcloned into the vector for extracellular expression. Western blotting analysis results showed that the PA-CPI was secreted into the medium. Bioassay demonstrated that E. coli DH5${\alpha}$ harboring pUC18ompAcat-CPI showed a significant difference in mortality to the Asian longhorned beetle Anoplophora glabripennis compared with untransformed E. coli DH5${\alpha}$ and control.

• Clinical and Experimental Vaccine Research
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• v.12 no.4
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• pp.304-312
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• 2023

Purpose: Due to the many problems with commercially available vaccines, the production of effective vaccines against brucellosis is a necessity. The aim of this study was to evaluate the immune responses caused by the chimeric protein consisting of trigger factor, Bp26, and Omp31 (TBO) along with aluminum hydroxide (AH/TBO) and selenium (Se/TBO) nanoparticles (NPs) as adjuvants in mouse model. Materials and Methods: Recombinant antigen expression was induced in Escherichia coli BL21 (DE3) bacteria using IPTG (isopropyl-d-1-thiogalactopyranoside). Purification and characterization of recombinant protein was conducted through NiFe₃O₄ NPs, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blot. NP characteristics, including morphology and particle size, were measured in vitro. The recombinant TBO was loaded on to AH and Se NPs and were administered subcutaneously. After mice immunization, measurement of antibody titter and protection assay was performed. Results: The average sizes of AH and Se NPs were about 60 nm and 150 nm, respectively. The enzyme-linked immunosorbent assay results showed that the serum of mice immunized by subcutaneous injection with both nanovaccines produced significant immunoglobulin G (IgG) responses against the chimeric antigen. The results of TBO-specific IgG isotype (IgG2a/IgG1) analysis showed that both AH and Se NPs induced a type to T-helper immune response. In addition, the results of the challenge with the pathogenic strain of Brucella melitensis 16M showed that vaccinated mice with AH/TBO NPs indicated a higher reduction of bacterial culture than immunized mice with Se/TBO NPs and TBO alone. Conclusion: The results showed that AH NPs carrying chimeric antigen can be a promising vaccine candidate against brucellosis by producing protective immunity.

• The Bulletin of The Korean Astronomical Society
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• v.42 no.2
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• pp.66.2-66.2
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• 2017

The Korea Astronomy and Space Science Institute plans to develop a coronagraph in collaboration with National Aeronautics and Space Administrative (NASA) and install it on the International Space Station (ISS). The coronagraph is an externally occulted one stage coronagraph with a field of view from 2.5 to 15 solar radii. The observation wavelength is approximately 400 nm where strong Fraunhofer absorption lines from the photosphere are scattered by coronal electrons. Photometric filter observation around this band enables the estimation of 2D electron temperature and electron velocity distribution in the corona. Together with the high time cadence (< 12 min) of corona images to determine the geometric and kinematic parameters of coronal mass ejections, the coronagraph will yield the spatial distribution of electron density by measuring the polarized brightness. For the purpose of technical demonstration, we intend to observe the total solar eclipse in 2017 August for the filter system and to perform a stratospheric balloon experiment in 2019 for the engineering model of the coronagraph. The coronagraph is planned to be installed on the ISS in 2021 for addressing a number of questions (e.g. coronal heating and solar wind acceleration) that are both fundamental and practically important in the physics of the solar corona and of the heliosphere.

• Journal of Applied Biological Chemistry
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• v.58 no.3
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• pp.227-232
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• 2015

A microarray study has been employed to understand changes of gene expression in E. coli KD43162 resistant to ampicillin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, cefazolin, cefepime, aztreonam, imipenem, meropenem, gentamicin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, fosfomycin, and trimethoprim-sulfamethoxazole except for amikacin using disk diffusion assay. Using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF MS analyses, 36 kDa of outer membrane proteins (OMPs) was found to be deleted in the multidrug resistant E. coli KD 43162. Microarray analysis was used to determine up- and down-regulated genes in relation to multidrug resistant E. coli KD43162. Among the up-regulated genes, these genes were corresponded to express the proteins as penicillin-binding proteins (PBPs), tartronate semialdehyde reductase, ethanolamine utilization protein, shikimate kinase I, allantoinase, predicted SAM-dependent methyltransferase, L-glutamine: D-fructose-6-phosphate aminotransferase (GFAT), phospho-glucosamine mutase, predicted N-acetylmannosamine kinase, and predicted N-acetylmannosamine-6-P epimerase. Up-regulation of PBPs, one of primary target sites of antibiotics, might be responsible for the multidrug resistance in E. coli with increasing amount of target sites. Up-regulation of GFAT enzyme may be related to the up-regulation of PBPs because GFAT produces N-acetylglucosamine, a precursor of peptidoglycans. One of GFAT inhibitors, azaserine, showed a potent inhibition on the growth of E. coli KD43162. In conclusion, up-regulation of PBPs and GFATs with the loss of 36 kDa OMP refers the multidrug resistance in E. coli KD 43162.

• Journal of Ginseng Research
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• v.45 no.1
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• pp.75-85
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• 2021

Background: Invasive infections due to foodborne pathogens, including Salmonella enterica serovar Typhimurium, are prevalent and life-threatening. This study aimed to evaluate the effects of ginsenoside Rg3 (Rg3) on the adhesion, invasion, and intracellular survival of S. Typhimurium. Methods: The impacts of Rg3 on bacterial growth and host cell viability were determined using the time kill and the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays, respectively. Gentamicin assay and confocal microscopic examination were undertaken to determine the effects of Rg3 on the adhesive and invasive abilities of S. Typhimurium to Caco-2 and RAW 264.7 cells. Quantitative reverse transcription polymerase chain reaction was performed to assess the expression of genes correlated with the adhesion, invasion, and virulence of S. Typhimurium. Results: Subinhibitory concentrations of Rg3 significantly reduced (p < 0.05) the adhesion, invasion, and intracellular survival of S. Typhimurium. Rg3 considerably reduced (p < 0.05) the bacterial motility as well as the release of nitrite from infected macrophages in a concentration-dependent manner. The expression of genes related to the adhesion, invasion, quorum sensing, and virulence of S. Typhimurium including cheY, hilA, OmpD, PrgK, rsgE, SdiA, and SipB was significantly reduced after Rg3 treatment. Besides, the compound downregulated rac-1 and Cdc-42 that are essential for actin remodeling and membrane ruffling, thereby facilitating Salmonella entry into host cells. This report is the first to describe the effects of Rg3 on "trigger" entry mechanism and intracellular survival S. Typhimurium. Conclusion: Rg3 could be considered as a supplement agent to prevent S. Typhimurium infection.

• Journal of KIISE:Information Networking
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• v.31 no.4
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• pp.384-392
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• 2004

Mobile contents service providers have some difficulties to timely and proper service deployment due to rapid development cycle of diverse portable devices with different capabilities. A way to resolve the problem is to introduce a mobile contents service paltform that can adapt original mobile contents to diverse devices dynamically and automatically. In this paper, we propose a mobile contents service platform based on active network mechanisms, called Mobile Content Adaptation Network(MobiCAN). The MobiCAN node provides effective service deployment, execution, and maintenance features and accommodates service layering and service customization capabilities for easy deployment. The basic functional units of the MobiCAN node are micro services with well-defined service interfaces and service layering features. For reliable services among the MobiCAN nodes, we design new distributed and robust Overlay Management Protocols(OMPs). As an example of practical MobiCAN applications, we finally describe Dynamic Contents Customization Proxy(DCCP) service.

• Korean Journal of Veterinary Research
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• v.50 no.4
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• pp.285-295
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• 2010

Protein expression patterns in Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) strains with diverse phenotypes, such as phage type, antibiotic resistance pattern and plasmid profiles were examined. For detailed analysis of proteins expressed by different S. Typhimurium strains, protein fractions were divided into detergent-rich phase (DP) and aqueous phase (AP) using triton X-114 detergent. The two phases were subjected to two-dimensional gel electrophoresis (2-DE), followed by protein identification using peptide mass fingerprinting (PMF). In the results, PMF showed that DP fractions consisted mainly of outer membrane proteins, whereas the AP fractions included cytosolic proteins. Comparison of 2-DE profiles of DP did not show any distinct protein spots which could be correlated with phage type, antibiotic resistance pattern or plasmid profile. However, comparisons of 2-DE profiles of the AP revealed differences in the protein spots, which could be correlated with the plasmid profile and phage types. Among these protein spots, flagellin was specific for strains containing a 90 kb plasmid. Compared to DT193 phage type, three protein spots in the range of pI 5.0-5.5 and MW 8-15 kDa of AP 2-DE profiles were absent in the DT104 phage types. Additionally, a protein spot with PI in the range of 4.5-5.0 and molecular weight (MW) between 51-69 kDa was specific for phage type DT104, while a protein spot with pI in the range of 4.0-4.8 and MW between 18-20 kDa was specific for DT193 phage type. These protein spots may be useful for discriminating phage types of S. Typhimurium.