• International Journal of Oral Biology
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• 제42권2호
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• pp.63-70
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• 2017

Selecting an appropriate antigen with optimal immunogenicity and physicochemical properties is a pivotal factor to develop a protein based subunit vaccine. Despite rapid progress in modern molecular cloning and recombinant protein technology, there remains a huge challenge for purifying and using protein antigens rich in hydrophobic domains, such as membrane associated proteins. To overcome current limitations using hydrophobic proteins as vaccine antigens, we adopted in silico analyses which included bioinformatic prediction and sequence-based protein 3D structure modeling, to develop a novel periodontitis subunit vaccine against the outer membrane protein FomA of Fusobacterium nucleatum. To generate an optimal antigen candidate, we predicted hydrophilicity and B cell epitope parameter by querying to web-based databases, and designed a truncated FomA (tFomA) candidate with better solubility and preserved B cell epitopes. The truncated recombinant protein was engineered to expose epitopes on the surface through simulating amino acid sequence-based 3D folding in aqueous environment. The recombinant tFomA was further expressed and purified, and its immunological properties were evaluated. In the mice intranasal vaccination study, tFomA significantly induced antigen-specific IgG and sIgA responses in both systemic and oral-mucosal compartments, respectively. Our results testify that intelligent in silico designing of antigens provide amenable vaccine epitopes from hard-to-manufacture hydrophobic domain rich microbial antigens.

• Molecules and Cells
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• 제41권6호
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• pp.545-552
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• 2018

Spleen tyrosine kinase (SYK) is a cytosolic non-receptor protein tyrosine kinase. Because SYK mediates key receptor signaling pathways involving the B cell receptor and Fc receptors, SYK is an attractive target for autoimmune disease and cancer treatments. To date, representative oral SYK inhibitors, including fostamatinib (R406 or R788), entospletinib (GS-9973), cerdulatinib (PRT062070), and TAK-659, have been assessed in clinical trials. Here, we report the crystal structures of SYK in complex with two newly developed inhibitors possessing 4-aminopyrido[4,3-D]pyrimidine moieties (SKI-G-618 and SKI-O-85). One SYK inhibitor (SKI-G-618) exhibited moderate inhibitory activity against SYK, whereas the other inhibitor (SKI-O-85) exhibited a low inhibitory profile against SYK. Binding mode analysis indicates that a highly potent SYK inhibitor might be developed by modifying and optimizing the functional groups that interact with Leu377, Gly378, and Val385 in the G-loop and the nearby region in SYK. In agreement with our structural analysis, one of our SYK inhibitor (SKI-G-618) shows strong inhibitory activities on the ${\beta}$-hexosaminidase release and phosphorylation of SYK/Vav in RBL-2H3 cells. Taken together, our findings have important implications for the design of high affinity SYK inhibitors.

• 동의생리병리학회지
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• 제23권6호
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• pp.1449-1459
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• 2009

Coptidis rhizoma (huanglian) is an herb that is widely used in traditional Chinese medicine that has recently been shown to possess anticancer activity. However, the molecular mechanism underlying the anticancer effects of this herb is poorly understood. In this study, we investigated the anticancer activity of a combination of CR extract and arsenic trioxide, as well as the apoptotic pathway associated with its mechanism of action in human lung cancer H157 cells. Combined treatment of H157 cells with CR extract and arsenic trioxide resulted in significant apoptotic death. In addition, combined treatment with CR extract and arsenic trioxide acted in concert to induce a loss of mitochondrial membrane potential (${\Delta}{\Psi}$), the release of cytochrome c from mitochondria, and an increase in the expression of pro-apoptotic p53 and Bax protein, which resulted in activation of caspases and apoptosis. CR extract combined with arsenic trioxide also increased the lipid peroxidation, mRNA expression of DR4 and DR5 and caspase-8 activity. These data indicate that combined treatment with CR extract and arsenic trioxide enhanced apoptotic cell death in H157 cells through diverse pathways, including mitochondrial dysfunction and death receptors, particularly DR4 and DR5. Thus, this treatment may be an effective from of chemotherapy.

• 한국한의학연구원논문집
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• 제17권3호
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• pp.77-85
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• 2011

Objectives : The purpose of this study was to develop a simulator which can analyze the anti-inflammatory effects of medical herbs based on e-cell concerning p38 MAPK signal pathway. Methods : We collected data concerning medical herbs with anti-inflammatory effects and the active compounds to provide as a fundamental databse and to validate the newly developed algorithm. At this time, we used the target database as pubmed and gathered the data by data mining tool, pathway studio. Also we have developed the web-based search system for confirming database related to anti-inflammation. We researched the mechanism of actions of proteins in p38 MAPK signal pathway when active compound has been inserted into the network. We reduced total network into TAK-MKK3-p38 and made the two types of mathematical model about active compounds' interaction. Results & Conclusion : We constructed the database which have 69 cases of medical herbs, 71 cases of active compounds, about 8,000 cases of URL(Uniform Resource Locator) related to papers and reports. We designed the ordinary differential equations for response of TAK, MKK3, p38 in e-cell's cytosol and nucleus. We used this formular as measure whether an active compound of medicinal plants which is inputted by an user would have an anti-inflammation effects. We developed the visualization program which could show the change of concentration over time.

• Journal of Welding and Joining
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• 제1권2호
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• pp.45-52
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• 1983

Many pressure vessels for the hot H$\sub$2//H$\sub$2/S service are made of 2+1/4Cr-1Mo steel with austenitic stainless steel overlay to combat agressive corrosion due to hydrogen sulfide. Hydrogen dissolves in to materials during operation, and sometimes gives rise to unfore-seeable damages. Appropriate precautions must, therefore, be taken to avoid the hydrogen induced damages in the design, fabrication and operation stage of such reactor vessels. Recently, hydrogeninduced cracking (or Disbonding) was found at the interface between base metal and stainless weld overlay of a desulfurizing reactor. Since the stainless steel overlay weld metal is subjected to thermal and internal-pressure loads in reactor operation, it is desirable for the overlay weld metal to have high strength and ductility from the stand point of structural safety. In section III of ASME Boiler and Pressure Vessel Code, Post-Weld Heat Treatment(PWHT) of more than one hour per inch at over 1100.deg. F(593.deg. C) is required for the weld joints of low alloy pressure vessel steels. This heat treatment to relieve stresses in the welded joint during construction of the pressure vessel is considered to cause sensitization of the overlay weld metal. The present study was carried out to make clear the diffusion of carbon migration by PWHT in dissimilar metal welded joint. The main conclusion reached from this study are as follows: 1) The theoretical analysis for diffusion of carbon in stainless steel overlay weld metal does not agree with Fick's 2nd law but the general law of molecular diffusion phenomenon by thermodynamic chemical potential. 2) In the stainless steel overlay welded joint, the PWHT at 720.deg. C for 10 hours causes a diffusion of carbon atoms from ferritic steel into austenitic steel according to the theoretical analysis for carbon migration and its experiment. 3) In case of PWHT at 720.deg. C for 10 hours, the micro-hardness of stainless steel weld metal in bonded zone increase very highly in the carburized layer with remarkable hardening than that of weld metal.

• Parasites, Hosts and Diseases
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• 제53권2호
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• pp.177-187
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• 2015

Merozoite surface proteins (MSPs) of malaria parasites play critical roles during the erythrocyte invasion and so are potential candidates for malaria vaccine development. However, because MSPs are often under strong immune selection, they can exhibit extensive genetic diversity. The gene encoding the merozoite surface protein-3 (MSP-3) of Plasmodium falciparum displays 2 allelic types, K1 and 3D7. In Thailand, the allelic frequency of the P. falciparum msp-3 gene was evaluated in a single P. falciparum population in Tak at the Thailand and Myanmar border. However, no study has yet looked at the extent of genetic diversity of the msp-3 gene in P. falciparum populations in other localities. Here, we genotyped the msp-3 alleles of 63 P. falciparum samples collected from 5 geographical populations along the borders of Thailand with 3 neighboring countries (Myanmar, Laos, and Cambodia). Our study indicated that the K1 and 3D7 alleles co-existed, but at different proportions in different Thai P. falciparum populations. K1 was more prevalent in populations at the Thailand-Myanmar and Thailand-Cambodia borders, whilst 3D7 was more prevalent at the Thailand-Laos border. Global analysis of the msp-3 allele frequencies revealed that proportions of K1 and 3D7 alleles of msp-3 also varied in different continents, suggesting the divergence of malaria parasite populations. In conclusion, the variation in the msp-3 allelic patterns of P. falciparum in Thailand provides fundamental knowledge for inferring the P. falciparum population structure and for the best design of msp-3 based malaria vaccines.

• 한국해양공학회지
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• 제27권5호
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• pp.105-114
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• 2013

Lots of orders of special vessels and offshore plants for developing the resources in deepwater have been increased in recent. Because the most of accidents on those structures are caused by fire and explosion, many researchers have been investigated quantitatively to predict the cause and effect of fire and explosion based on both experiments and numerical simulations. The first step of the evaluation procedures leading to fire and explosion is to predict the dispersion of flammable or toxic material, in which the released material mixes with surrounding air and be diluted. In particular turbulent mixing, but density differences due to molecular weight or temperature as well as diffusion will contribute to the mixing. In the present paper, the numerical simulation of hydrogen dispersion inside a simple-shaped offshore structure was performed using a commercial CFD program, ANSYS-CFX. The simulated results for concentration of released hydrogen are compared to those of experiment and other simulation in Jordan et al.(2007). As a result, it is seen that the present simulation results are closer to the experiments than other simulation ones. Also it seems that the hydrogen dispersion is closely related to turbulent mixing and the selection of the turbulence model properly is significantly of importance to the reproduction of dispersion phenomena.

• 대한화장품학회지
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• 제36권1호
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• pp.17-32
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• 2010

본 총설에서는 이산화탄소에 용해력이 있는 새로운 탄화수소 공중합체의 설계와 개발, 그리고 생체친화성 고분자의 초임계 중합을 위한 효과적인 계면활성제로써의 성능에 대해 소개하고, 초입계 유체의 기본적 개념을 용매로서의 성질과 고분자 합성분야에서의 응용적인 측면에서 기술한다. 이산화탄소에 높은 용해력을 지닌 탄화수소 고분자 중합을 위해 새로운 리빙라디칼 중합기술을 사용하였고, 이 물질들의 이산화탄소 내에서의 상거동을 측정하여 공중합체의 분자량과 구조가 용해도에 미치는 영향을 조사하였다. 초임계 분산중합에서의 효과적인 계면활성력을 확인하였고, 성장하는 입자의 안정화에 필요한 키 파라미터를 결정하기 위해 다양한 조건에서 실험을 수행하였으며, 화장품 분야에 응용될 수 있는 새로운 구조의 친환경 고분자 소재 개발에 이 연구가 작용될 수 있다는 잠재적인 가능성을 확인하였다.

• 한국미생물·생명공학회지
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• 제49권2호
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• pp.181-191
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• 2021

The present study addresses isolation, optimization, partial purification, and characterization of a haloalkaline serine protease from a newly isolated haloarchaeal strain isolated from Wadi El Natrun in Egypt. We expected that a two-step sequential statistical approach (one variable at a time, followed by response surface methodology) might maximize the production of the haloalkaline serine protease. The enzyme was partially purified using Hiprep 16/60 sephacryl S-100 HR gel filtration column. Molecular identification revealed the newly isolated haloarchaeon to be Natrialba hulunbeirensis strain WNHS14. Among several tested physicochemical determinants, casamino acids, KCl, and NaCl showed the most significant effects on enzyme production as determined from results of the One-Variable-At-A-time (OVAT) study. The BoxBehnken design localized the optimal levels of the three key determinants; casamino acids, KCl, and NaCl to be 0.5% (w/v), 0.02% (w/v), and 15% (w/v), respectively, obtaining 62.9 U/ml as the maximal amount of protease produced after treatment at 40℃, and pH 9 for 9 days with 6-fold enhancement in yield. The enzyme was partially purified after size exclusion chromatography with specific activity, purification fold, and yield of 1282.63 U/mg, 8.9, and 23%, respectively. The enzyme showed its maximal activity at pH, temperature, and NaCl concentration optima of 10, 75℃, and 2 M, respectively. Phenylmethylsulfonyl fluoride (PMSF, 5 mM) completely inhibited enzyme activity.

• Journal of Microbiology and Biotechnology
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• 제31권4호
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• pp.501-509
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• 2021

More than half the world's population is thought to be infected with Helicobacter pylori. Although the majority of infected people are asymptomatic, H. pylori infection may cause gastric ulcers and deadly gastric cancer. Owing to the difficulty and invasiveness of current routine culture and diagnostic methods, a highly sensitive and specific noninvasive assay for H. pylori is of interest. This study highlighted the design and performance of a colorimetric magneto loop-mediated isothermal amplification (CM-LAMP) assay to detect H. pylori in spiked saliva samples. LF primers were coated on magnetic nanoparticles by carbodiimide-induced immobilization and functionally used for solid-phase amplification. During the LAMP reaction at 66℃, biotin-tagged FIPs were incorporated into LAMP amplicons. The colorimetric signal developed after the addition of NeutrAvidin horseradish peroxidase conjugate (NA-HRP) and ABTS. None of the tested microorganisms, including closely related bacteria, was shown positive by the CM-LAMP assay except H. pylori isolates. This novel platform was highly specific and 100-fold more sensitive (40 CFU/ml or 0.2 CFU per reaction) than the PCR and conventional LAMP assays for the detection of H. pylori in spiked saliva. Our results demonstrated the feasibility of using this noninvasive molecular diagnostic test to detect H. pylori in saliva samples.