• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1810-1818
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• 2015

For the synthesis of various pharmaceuticals, chiral alcohols are useful intermediates. Among them, (R)-ethyl-4-chloro-3-hydroxybutanoate ((R)-ECHB) is an important building block for the synthesis of L-carnitine. (R)-ECHB is produced from ethyl-4-chloro-3-oxobutanoate (ECOB) by a reductase-mediated, enantioselective reduction reaction. The Saccharomyces cerevisiae YOL151W reductase that is expressed in Escherichia coli cells exhibited an enantioselective reduction reaction toward ECOB. By virtue of the C-terminal His-tag, the YOL151W reductase was purified from the cell-free extract using Ni²⁺-NTA column chromatography and immobilized onto Ni²⁺-magnetic microparticles. The physical properties of the immobilized reductase (Imm-Red) were measured using electron microscopy, a magnetic property measurement system, and a zeta potential system; the average size of the particles was approximately 1 μm and the saturated magnetic value was 31.76 emu/g. A neodymium magnet was used to recover the immobilized enzyme within 2 min. The Imm-Red showed an optimum temperature at 45℃ and an optimum pH at 6.0. In addition, Bacillus megaterium glucose dehydrogenase (GDH) was produced in the E. coli cells and was used in the coupling reaction to regenerate the NADPH cofactor. The reduction/oxidation coupling reaction composed of the Imm-Red and GDH converted 20 mM ECOB exclusively into (R)-ECHB with an e.e._p value of 98%.

• The Journal of Korean Academy of Prosthodontics
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• v.47 no.3
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• pp.348-355
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• 2009

Statement of problem: Although the use of colorimeter to improve the accuracy of tooth color measurement has increased, it is real situation that there is almost no comparative study about the accuracy among various colorimeters. Purpose: The purpose of this study is to compare the accuracy of porcelain color reproducibility using ShadeEye-Ncc dental chroma meter (Shofu Inc., Kyoto, Japan) and Shadepilot$^{TM}$ system (Degudent Inc., Hanau, Germany). Material and methods: Color of 2M2 and 3M2 shade tap of Vitapan 3D Master shade guide were measured using the above two colorimeters. Porcelain specimens were fabricated through this data. The porcelain systems used were Vintage Halo Porcelain system with ShadeEye-Ncc dental chroma meter, and VitaOmega900 Porcelain system with Shadepilot$^{TM}$ system. Colors of shade tap models and porcelain specimen were measured with spectrophotometer, and calculated delta E based on the results. Results: The following results were obtained. 1. Shadepilot$^{TM}$ system showed more suitable porcelain color reproducibility compared to ShadeEye-Ncc dental chroma meter (P <.05). 2. The result showed the difference of porcelain color reproducibility according to shade of Shade tap which is model. More suitable porcelain color reproducibility was shown at 3M2 shade tap than at 2M2 shade tap (P <.05). 3. The average of ${\Delta}E*$ of porcelain specimens which was made by the two colorimeters was 4.44-6.14, so the difference of shade was shown through the naked eye. Conclusion: Shadepilot$^{TM}$ system showed more suitable porcelain color reproducibility. When using ShadeEye-Ncc dental chromameter, it will be necessary to improve accuracy through coloring various parts of the whole tooth.

• Transactions of the Korean Society of Mechanical Engineers
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• v.16 no.4
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• pp.649-657
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• 1992

Transmission error for a stepping motor-timing belt drive system is investigated experimentally and analytically. From FFT analysis of the experimental results, it is found that the transmission error consists of three periodic errors : (1) error by the stepping motor per one resolution angle theta.$_{m}$, (2) error by the pulley eccentricity per one revolution theta.$_{e}$, and (3) error by the meshing effect between the belt and the pulley teeth per one pitch revoltion theta.$_{p}$. In order to investigate the effects of some design parameters on the transmission error, the dynamic models of the stepping motor-timing belt drive system are derived by Bondgraph. According to the simulation results, as the belt total tension increases, theta.$_{m}$ and theta.$_{e}$ decrease due to the nonlinearity of the belt. In adition, the numerical and experimental results show that theta.$_{m}$ and theta.$_{e}$ of the loaded case are larger than those of the unloaded case. The analytical results are in good accordance with the experimental results.sults.s.sults.

• KSBB Journal
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• v.24 no.4
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• pp.341-346
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• 2009

Candida Antarctica lipase A (CalA) has been used because of its suitability in industrial applications. CalA has unique features capable to accept tertiary and sterically hindered alcohols among many hydrolases. CalA gene was cloned and constructed in expression vector such as pColdIII/CalA and $pPICZ{\alpha}A$/CalA. The gene encoding pColdIII/CalA was functionally expressed in the cytoplasm of Escherichia coli $Origami^{TM}$ B (DE3) cells. The plasmid $pPICZ{\alpha}A$/CalA linearized by BstX I was integrated into 5'AOX1 region of the chromosomal DNA and was functionally expressed in the methyl atrophic yeast Pichia pastoris. Expressed CalA in P. pastoris (0.7 Unit/mL) showed 35 times higher activity than that in E. coli expression system (0.02 Unit/mL).

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.2
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• pp.144-149
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• 2001

For the production and purification of a single chain human insulin precursor, four types of fusion peptides $\beta$-galactosidase (LacZ), maltose binding protein (MBP), glutathione-S-transferase (GST), and (His)(sub)6-tagged sequence (HTS) were investigated. Recombinant E. coli harboring hybrid genes was cultivated at 37$\^{C}$ for 1h, and gene induction occurred when 0.2mM of isopropyl-D-thiogalactoside (IPTG) was added to the culture broth, except for E. coli BL21 (DE3) pLysS harboring a pET-BA cultivation with 1.0mM IPTG, followed by a longer than 4h batch fermentation respectively. DEAE-Sphacel and Sephadex G-200 gel filtration chromatography, amylose affinity chromatography, glutathione-sepharose 4B affinity chromatography, and a nickel chelating affinity chromatography system as a kind of immobilized metal ion affinity chromatography (IMAC) were all employed for the purification of a single chain human insulin precursor. The recovery yields of the HTS-fused, GST-fused, MBP-fused, and LacZ-fused single chain human insulin precursors resulted in 47%, 20%, 20%, and 18% as the total protein amounts respectively. These results show that a higher recovery yield of the finally purified recombinant peptides was achieved when affinity column chromatography was employed and when the fused peptide had a smaller molecular weight. In addition the pET expression system gave the highest productivity of a fused insulin precursor due to a two-step regulation of the gene expression, and the HTS-fused system provided the highest recovery of a fused insulin precursor based on a simple and specific separation using the IMAC technique.

• The Journal of Pediatrics of Korean Medicine
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• v.34 no.4
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• pp.22-30
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• 2020

Objectives This study was conducted to confirm the inhibitory effect of β-glucan on epithelial inflammation induced by atopic dermatitis through Endocannabinoid system (ECS) activity. Methods Six-week-old NC/Nga mice were divided into a control group (Ctrl), atopic dermatitis elicitation group (ADE), and a β-glucan-treated group (β-glucan treatment after atopy dermatitis elicitation, β-GT). After 3 weeks, CB1, CB2, and GPR55 were observed to confirm the regulation of ECS activity, and filaggrin in the stratum corneum and Kallikrein-related peptidase (KLK) 7 in the stratum corneum and protease activated receptor (PAR)-2 were observed to confirm the inhibition of the inflammation, Phosphorylated extracellular signal-related kinase (p-ERK), Phosphorylated mammalian target of rapamycin (p-mTOR), and E-Cadherin were observed to confirm microenvironmental regulation. Results β-GT was significantly increased in CB1, CB2, and GPR55 positive reactions compared to that of the ADE. In positive reaction of the filaggrin in the stratum corneum, β-GT was significantly increased than that of the ADE. For KLK7 positive and PAR2 positive, β-GT was significantly reduced compared to the ADE. The p-ERK-positive and p-mTOR-positive reactions were significantly reduced in β-GT than in ADE. E-cadherin positive reaction was significantly increased in β-GT than in ADE (All p < 0.01). Conclusions It was confirmed that β-glucan has the effect of inhibiting the epithelium induced by atopic dermatitis through the ECS activity.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.2
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• pp.155-162
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• 2020

Equine follicle stimulating hormone receptor (eFSHR) has a large extracellular domain and an intracellular domain containing approximately 10 phosphorylation sites within the G protein-coupled receptor. This study was conducted to analyze the function of phosphorylation sties at the eFSHR C-terminal region. We constructed a mutant of eFSHR, in which the C-terminal cytoplasmic tail was truncated at residue 641 (eFSHR-t641). This removed 10 potential phosphorylation sites from the C-terminal region of the intracellular loop. The eFSHR-wild type (eFSHR-wt) and eFSHR-t641 cDNAs were subcloned into the pCMV-ARMS1-PK2 expression vector. These plasmids were transfected into PathHunter CHO-K1 Parental cells expressing β-arrestin 2 enzyme acceptor fusion protein and analyzed for agonist-induced cAMP response. The cAMP response in cells expressing eFSHR-t641 was lower than the response in cells expressing eFSHR-wt. EC₅₀ values of eFSHR-wt and eFSHR-t641 were 1079 ng/mL and 1834 ng/mL, respectively. eFSHR-t641 was approximately 0.58-fold compared with that of eFSHR-wt. The maximal response in eFSHR-wt and eFSHR-t641 was 24.7 nM and 16.7 nM, respectively. The Rmax value of phosphorylation sites in eFSHR-t641 was also decreased to approximately 68.4% of that in eFSHR-wt. The collective data implicate that the phosphorylation sites in the eFSHR C-terminal region have a pivotal role in signal transduction in PathHunter CHO-K1 cells, and indicate that β-arrestin is involved in coupling the activated receptors to the internalization system.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.21 no.12
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• pp.2261-2269
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• 2017

The current LTE network system provides service by locally allocating eNodeB to the EPC(Evolved Packet Core). However, it is not suitable for inter-node communication based on the distributed network environment. In this paper, we propose an integrated system by configuring E-UTRAN and EPC as All-In-One type to enable service in each distributed node. M2M UE is mounted on a each node for wireless In-band backhaul link. The integrated node provides inter-node communication over the wireless in-band backhaul link. If a normal UE moves and an access node changes in the system all entities of LTE integrated into one, it is necessary to change the P-GW which is IP anchor. In order to support the inter-node mobility even if P-GW is changed, We defined UPFE(User Packet Forwarding Extension) scheme and implemented the handover procedure of EPC. Also, we analyze the cell range of the integrated LTE system with the addition of the node and compare the handover delay with the current system.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.6
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• pp.511-519
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• 2001

Nimopidine, one of dihydropyridine derivatives, has been widely used to pharmacologically identify L-type Ca currents. In this study, it was tested if nimodipine is a selective blocker for L-type Ca currents in sensory neurons and heterologous system. In mouse dorsal root ganglion neurons (DRG), low concentrations of nimodipine $(<10\;{\mu}M),$ mainly targeting L-type Ca currents, blocked high-voltage-activated calcium channel currents by ${\sim}38%.$ Interestingly, high concentrations of nimodipine $(>10\;{\mu}M)$ further reduced the 'residual' currents in DRG neurons from ${\alpha}_{1E}$ knock-out mice, after blocking L-, N- and P/Q-type Ca currents with $10\;{\mu}M$ nimodipine, $1\;{\mu}M\;{\omega}-conotoxin$ GVIA and 200 nM ${\omega-agatoxin$ IVA, indicating inhibitory effects of nimodipine on R-type Ca currents. Nimodipine $(>10\;{\mu}M)$ also produced the inhibition of both low-voltage-activated calcium channel currents in DRG neurons and ${\alpha}_{1B}\;and\;{\alpha}_{1E}$ subunit based Ca channel currents in heterologous system. These results suggest that higher nimodipine $(>10\;{\mu}M)$ is not necessarily selective for L-type Ca currents. While care should be taken in using nimodipine for pharmacologically defining L-type Ca currents from native macroscopic Ca currents, nimodipine $(>10\;{\mu}M)$ could be a useful pharmacological tool for characterizing R-type Ca currents when combined with toxins blocking other types of Ca channels.

• Korean Journal of Optics and Photonics
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• v.4 no.1
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• pp.101-107
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• 1993

We carried out the useful theoretical calculation for the optimum design of a 1.48 ${\mu}m$ diode laser pumped E$r^{3+}$ doped fiber amplifier. The model we established is based on the rate equations of three level laser system and the overlap integral between fundamental mode L$P_{01}$ and E$r^{3+}$ doped area. We determined several fiber parameters (N.A., V value, fiber length, E$r^{3+}$ concentration, cutoff wavelength etc.) for the optimum design of a high optical gain. We found that our theoretical results are very useful to the design of E$r^{3+}$ doped fiber used in EDFA.