• Biomolecules & Therapeutics
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• v.23 no.2
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• pp.189-194
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• 2015

P450 1A2 is responsible for the metabolism of clinically important drugs and the metabolic activation of environmental chemicals. Genetic variations of P450 1A2 can influence its ability to perform these functions, and thus, this study aimed to characterize the functional significance of three P450 1A2 allelic variants containing nonsynonymous single nucleotide polymorphisms (P450 $1A2^*8$, R456H; $^*15$, P42R; $^*16$, R377Q). Variants containing these SNPs were constructed and the recombinant enzymes were expressed and purified in Escherichia coli. Only the P42R variant displayed the typical CO-binding spectrum indicating a P450 holoenzyme with an expression level of ~ 170 nmol per liter culture, but no P450 spectra were observed for the two other variants. Western blot analysis revealed that the level of expression for the P42R variant was lower than that of the wild type, however the expression of variants R456H and R377Q was not detected. Enzyme kinetic analyses indicated that the P42R mutation in P450 1A2 resulted in significant changes in catalytic activities. The P42R variant displayed an increased catalytic turnover numbers ($k_{cat}$) in both of methoxyresorufin O-demethylation and phenacetin O-deethylation. In the case of phenacetin O-deethylation analysis, the overall catalytic efficiency ($k_{cat}/K_m$) increased up to 2.5 fold with a slight increase of its $K_m$ value. This study indicated that the substitution P42R in the N-terminal proline-rich region of P450 contributed to the improvement of catalytic activity albeit the reduction of P450 structural stability or the decrease of substrate affinity. Characterization of these polymorphisms should be carefully examined in terms of the metabolism of many clinical drugs and environmental chemicals.

• Korean Journal of Veterinary Research
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• v.54 no.4
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• pp.209-218
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• 2014

Experimental autoimmune encephalomyelitis (EAE), an animal model of human multiple sclerosis (MS), reflects pathophysiologic steps in MS such as the influence of T cells and antibodies reactive to the myelin sheath, and the cytotoxic effect of cytokines. Galectin-9 (Gal-9) is a member of animal lectins that plays an essential role in various biological functions. The expression of Gal-9 is significantly enhanced in MS lesions; however, its role in autoimmune disease has not been fully elucidated. To identify the role of Gal-9 in EAE, we measured changes in mRNA and protein expression of Gal-9 as EAE progressed. Expression increased with disease progression, with a sharp rise occurring at its peak. Gal-9 immunoreactivity was mainly expressed in astrocytes and microglia of the central nervous system (CNS) and macrophages of spleen. Flow cytometric analysis revealed that $Gal-9^+CD11b^+$ cells were dramatically increased in the spleen at the peak of disease. Increased expression of tumor necrosis factor (TNF)-R1 and p-Jun N-terminal kinase (JNK) was observed in the CNS of EAE mice, suggesting that TNF-R1 and p-JNK might be key regulators contributing to the expression of Gal-9 during EAE. These results suggest that identification of the relationship between Gal-9 and EAE progression is critical for better understanding Gal-9 biology in autoimmune disease.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.14 no.11
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• pp.2535-2540
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• 2010

In traditional Publish/Subscribe system, the first procedure to deliver event from publisher to subscriber is that publisher publishes publisher's event to broker. Next step is that broker checks simple binary notion of matching : an event either matches a subscription or it does not. Lastly, broker delivers the event matched with subscriptions to the corresponding subscribers. In this system, information delivery has been accomplished in one way only. However, current some applications require two way delivery between subscriber and publisher. Therefore, we initiate an extended Publish/Subscribe system that supports two way delivery. Extended Publish/Subscribe system requires additional functions of delivering subscription to publisher and especially deciding top-n subscriptions using priority because broker might has a number of subscriptions. In this paper, we propose two priority retrieval techniques based on SIF using IS-List with deciding priority among subscriptions and defining SIF(Specific Interval First). The performance measurements show that RSO(resulting set sorting) technique results in better performance in index creation time and ITS&IS(insertion time sorting and inverse search using stack) technique results in better performance in search time.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.34 no.10
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• pp.1427-1435
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• 2010

A mixed finite element model was developed using the classical plate theory to analyze the nonlinear bending of a plate. The appropriate weight functions for the constraints integrated over the domain were determined by the Lagrange multiplier method by using the principle of minimum virtual energy; which provides the constitutive relations between force-like variables and strains. All of detail terms of element wise coefficient matrices and associate tangent matrices to be used in the Newton iterative method are presented. Then, the linear solutions of the current model and those of the traditional displacement model under the SS (simple support) boundary conditions were compared with the existing analytical solution. The post-processed images of the nonlinear results of the force-like variables are presented to show the continuity of the solutions at the joint of the element boundaries. Finally, the converged nonlinear finite element solutions of the current model are compared with those of existing traditional displacement model.

• Earthquakes and Structures
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• v.2 no.3
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• pp.207-232
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• 2011

The development of reliable earthquake mitigation plans and seismic risk management procedures can only be based on the establishment of comprehensive earthquake hazard and loss scenarios. Two cities, Grevena (Greece) and D$\ddot{u}$zce (Turkey), were used as case studies in order to apply a comprehensive methodology for the vulnerability and loss assessment of lifelines. The methodology has the following distinctive phases: detailed inventory, identification of the typology of each component and system, evaluation of the probabilistic seismic hazard, geotechnical zonation, ground response analysis and estimation of the spatial distribution of seismic motion for different seismic scenarios, vulnerability analysis of the exposed elements at risk. Estimating adequate earthquake scenarios for different mean return periods, and selecting appropriate vulnerability functions, expected damages of the water and waste water systems in D$\ddot{u}$zce and of the roadway network and waste water system of Grevena are estimated and discussed; comparisons with observed earthquake damages are also made in the case of D$\ddot{u}$zce, proving the reliability and the efficiency of the proposed methodology. The results of the present study constitute a sound basis for the development of efficient loss scenarios for lifelines and infrastructure facilities in seismic prone areas. The first part of this paper, concerning the estimation of the seismic ground motions, has been utilized in the companion paper by Kappos et al. (2010) in the same journal.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.1
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• pp.61-66
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• 2011

P2Y receptors are metabotropic G-protein-coupled receptors, which are involved in many important biologic functions in the central nervous system including retina. Subtypes of P2Y receptors in retinal tissue vary according to the species and the cell types. We examined the molecular and pharmacologic profiles of P2Y purinoceptors in retinoblastoma cell, which has not been identified yet. To achieve this goal, we used $Ca^{2+}$ imaging technique and western blot analysis in WERI-Rb-1 cell, a human retinoblastoma cell line. ATP ($10\;{\mu}M$) elicited strong but transient $[Ca^{2+}]_i$ increase in a concentration dependent manner from more than 80% of the WERI-Rb-1 cells (n=46). Orders of potency of P2Y agonists in evoking $[Ca^{2+}]_i$ transients were 2MeS-ATP>ATP>>UTP=${\alpha}{\beta}$-MeATP, which was compatible with the subclass of $P2Y_1$ receptor. The $[Ca^{2+}]_i$ transients evoked by applications of 2MeS-ATP and/or ATP were also profoundly suppressed in the presence of $P2Y_1$ selective blocker (MRS 2179; $30\;{\mu}M$). $P2Y_1$ receptor expression in WERI-Rb-1 cells was also identified by using western blot. Taken together, $P2Y_1$ receptor is mainly expressed in a retinoblastoma cell, which elicits $Ca^{2+}$ release from internal $Ca^{2+}$ storage sites via the phospholipase C-mediated pathway. $P2Y_1$ receptor activation in retinoblastoma cell could be a useful model to investigate the role of purinergic $[Ca^{2+}]_i$ signaling in neural tissue as well as to find a novel therapeutic target to this lethal cancer.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.3
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• pp.177-183
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• 2010

Tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide synthase (NOS) activity, is known to play important roles in modulating both NO and superoxide production during vascular diseases such as atherosclerosis. However, the role of BH4 in functions of vascular smooth muscle cells is not fully known. In this study, we tested the effects of BH4 and dihydrobiopterin (BH2), a BH4 precursor, on migration and proliferation in response to platelet-derived growth factor-BB (PDGF-BB) in rat aortic smooth muscle cells (RASMCs). Cell migration and proliferation were measured using a Boyden chamber and a 5-bromo-2'-deoxyuridine incorporation assay, respectively, and these results were confirmed with an ex vivo aortic sprout assay. Cell viability was examined by 2,3-bis [2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide assays. BH4 and BH2 decreased PDGF-BBinduced cell migration and proliferation in a dose-dependent manner. The inhibition of cell migration and proliferation by BH4 and BH2 was not affected by pretreatment with $N^G$-nitro-L-arginine methyl ester, a NOS inhibitor. Moreover, the sprout outgrowth formation of aortic rings induced by PDGF-BB was inhibited by BH4 and BH2. Cell viability was not inhibited by BH4 and BH2 treatment. The present results suggest that BH4 and BH2 may inhibit PDGF-stimulated RASMC migration and proliferation via the NOS-independent pathway. Therefore, BH4 and its derivative could be useful for the development of a candidate molecule with an NO-independent anti-atherosclerotic function.

• Journal of Korean Medical classics
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• v.13 no.2 s.17
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• pp.21-21
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• 2000

All living things including human being consist of soul(spirit) and body. Soul is the root of a life and body frames it. I wrote this paper to tell how internal injury due to endogenous etiological factors and affection due to exegenous pathogenic factors, affect the internal organs and the external bodily from. This paper begins with description of the patterns of disharmony of the internal organs. General disorders of each Five-Jang(五藏) an be classified into two types of soul and body. The Liver and the Heart which lead changing to Yang(陽) of vital energy, have close relationship with spiritual symptoms because spirit is related to Yang. The Lung, Kidneys, the Spleen which lead changing to m(陰) of vital energy, mainly connected with physical disorders because body is relatively close to m. The Five-Jang are ruled by the Five-Phases(五行) system and cause troubles with Oche(五體) and the nine body orifice. Otherwise the main function of the Six-Bu(六府) is to receive food, absorb the usuable portions, and transmit and excrete waste. Therefore they can cause such problems as abdominal pain, distention, difficulty in urination, and constipation. The spleen is responsible for sending Grain-Ki)穀氣) 내 that is closely connected with the six-Bu. The Gall bladder takes care of control of giving out spirit. That's why it presents many symptoms related to the spirit that is ruled by the Five-Jang. Patterns of disharmony of external bodily form is influenced by the state of Maridians. Bodily forms get divided into many parts by the function of six-meridians(六經) to which they belong. Six-meridians have their own function related to excretion, related to excretion, retention, and balance(開闔樞). If local bodily froms get affected by pernicious influences, the Meridians to which they are attached will lose harmony and connot fulfil their own functions. Because the meridian symtem unifies all parts of the body, the whole body will be under the influence of the affection although local parts of the bodily forms get affected. Therefore the symptoms of disharmony should be considered in a synthetic view. There are however, also cases which should be focused on the very meridian channels affected. Disorder within a Meridian generates derangement in the pathway make it not to benefit the muscles and skin belong to it. Because the meridians connect the interior organs with the exterior bodily form, they are influenced by each other.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.140-144
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• 2003

Type Ⅰ signal peptidase is an integral membrane protein that functions to cleave signal peptides from secreted and membrane proteins. The enzyme serves as a potential target for the development of novel antibacterial agents due to its unique physiological properties. Despite being one of the best characterized enzymes, the catalysis of Type Ⅰ signal peptidase still remains controversy over the catalytic serine/lysine dyad mechanism. It appears that the dyad proteases are generally less efficient than the prototypical serine/histidine/aspartic acid triad found in most enzymes, although Type Ⅰ signal peptidase is an exception to this rule. In this paper, we have proposed that Type Ⅰ signal peptidase may act as the serine/lysine/aspartic acid triad cataltytic mechanism. Therefore, the aspartic acid 99 residue in the E. coli signal peptidase was chosen and mutated to an alanine to see if there is any possible role of the aspartic acid in the catalytic function. Type Ⅰ signal peptidase D99A protein was inactive in vitro assay using the procoat synthesized by in vitro transcription translation. However, the mutant was active using a highly sensitive in vivo assay. Pulse-chase experiments show that the replacement of aspartic acid 99 with alanine results in a very unstable signal peptidase molecule. Therefore, we conclude that it is unlikely that the residue is directly involved in catalysis, but rather plays an important role in stabilizing the protein structure.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.152-161
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• 2015

CodY is a highly conserved protein in low G+C gram-positive bacteria that regulates genes involved in sporulation and stationary-phase adaptation. Bacillus thuringiensis is a grampositive bacterium that forms spores and parasporal crystals during the stationary phase. To our knowledge, the regulatory mechanism of CodY in B. thuringiensis is unknown. To study the function of CodY protein in B. thuringiensis, BMB171codY^- was constructed in a BMB171 strain. A shuttle vector containing the ORF of cry1Ac10 was transformed into BMB171 and BMB171codY^-, named BMB171cry1Ac and BMB171codY^-cry1Ac, respectively. Some morphological and physiological changes of codY mutant BMB171codY^-cry1Ac were observed. A comparative proteomic analysis was conducted for both BMB171codY^-cry1Ac and BMB171cry1Ac through two-dimensional gel electrophoresis and MALDI-TOF-MS/MS analysis. The results showed that the proteins regulated by CodY are involved in microbial metabolism, including branched-chain amino acid metabolism, carbohydrate metabolism, fatty acid metabolism, and energy metabolism. Furthermore, we found CodY to be involved in sporulation, biosynthesis of poly-β-hydroxybutyrate, growth, genetic competence, and translation. According to the analysis of differentially expressed proteins, and physiological characterization of the codY mutant, we performed bacterial one-hybrid and electrophoretic mobility shift assay experiments and confirmed the direct regulation of genes by CodY, specifically those involved in metabolism of branched-chain amino acids, ribosomal recycling factor FRR, and the late competence protein ComER. Our data establish the foundation for in-depth study of the regulation of CodY in B. thuringiensis, and also offer a potential biocatalyst for functions of CodY in other bacteria.