• Journal of Biomedical Engineering Research
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• v.37 no.5
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• pp.186-194
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• 2016

In this study, a co-axial flow induced microfluidic chip to fabricate pure collagen type I microfiber via the control of collagen type I and Na-alginate gelation process. The pure collagen type I microfiber was generated by selective degradation of Ca-alginate from 'Core-Shell' structured hydrogel microfiber. To make 'Core-Shell' structure, collagen type I solution was introduced into core channel and 1.5% Na-alginate solution was injected into side channel in microfluidic chip. To evaluatethe 'Core-Shell' structure, the red and green fluorescence substances were mixed into collagen type I and Na-alginate solution, respectively. The fluorescence substances were uniformly loaded into each fiber, and the different fluorescence images were dependent on their location. By immoblizing EpH4-Ras and C6 cells within collagen type I and Na-alginate solution, we sucessfully demonstrated the co-culture of EpH4-Ras and C6 cells with 'Core-Shell' like hydrogel microfiber for 5 days. Only to produce pure collagen type I hydrogel fiber, tri-sodium citrate solution was used to dissolve the shell-like Ca-alginate hydrogel fiber from 'Core-Shell' structured hydrogel microfiber, which is an excellent advantage when the fiber is employed in three-dimensional scaffold. This novel method could apply various application in tissue engineering and biomedical engineering.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.979-985
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• 2001

A fluorometric detection technique for HDL (High Density Lipoprotein) and LDL (Low Density Lipoprotein) was developed for application in a fiber-optic immunosensor using a protein A Langmuir-Blodgget (LB) film. For the fluorescence immunoassay, antibodies specific to HDL or LDL were imobilied on the protein A LB film, and a fluorescence amplification method was developed to overcome their weak fluorescence. The deposition of protein A using the LB technique was monitored using a surface pressure-are $({\pi}-A)$ curve, and the antibody immobilization of the protein A LB film was experimentally verified. The immobilized antibody was used to separate only HDL and LDL from a sample, then the fluorescence of he separated HDL or LDL was amplified. The amount of LDL or HDL was measured using the developed fiber optic fluorescence detection system. The optical properties resulting from the reaction of HDL or LDL with o-phtaldialdehyde, detection range, response time, and stability of the immunoassay were all investigated. The respective detection ranges for HDL and LDL were sufficient to diagnose the risk of coronary heart disease. The amplification step increased the sensitivity, while selective separation using the immobilized antibody led to linearity in the sensor signal. The regeneration of the antibody-immobilized substrate could produce a stable and reproducible immunosensor.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1634-1639
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• 2015

Cytochrome P450 hydroxylase (CYP) in actinomycetes plays an important role in the biosynthesis and bioconversion of various secondary metabolites. Two unique CYPs named CYP-sb21 and CYP-pa1, which were identified from Sebekia benihana and Pseudonocardia autotrophica, respectively, were proven to transfer a hydroxyl group at the 4^th or 9^th N-methyl leucine position of immunosuppressive agent cyclosporin A (CsA). Interestingly, these two homologous CYPs showed different CsA regio-selectivities. CYP-sb21 exhibited preferential hydroxylation activity at the 4^th position over the 9^th position, whereas CYP-pa1 showed the opposite preference. To narrow down the CYP domain critical for CsA regio-selectivity, each CYP was divided into four domains, and each domain was swapped with its counterpart from the other CYP. A total of 18 hybrid CYPs were then individually tested for CsA regio-selectivity. Although most of the hybrid CYPs failed to exhibit a significant change in regio-selectivity in the context of CsA hydroxylation, hybrid CYP-pa1 swapped with the second domain of CYP-sb21 showed a higher preference for the 9^th position. Moreover, hybrid CYPsb21 containing seven amino acids from the 2nd domain of CYP-pa1 showed higher preference for the 4^th position. These results imply that the 2nd domain of CsA-specific CYP plays a critical role in CsA regio-selectivity, thereby setting the stage for biotechnological application of CsA regio-selective hydroxylation.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.911-917
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• 2009

The apoptotic caspases have been classified in accordance with their substrate specificities, as the optimal tetrapeptide recognition motifs for a variety of caspases have been determined via positional scanning substrate combinatorial library technology. Here, we focused on two proteolytic recognition motifs, DEVD and IETD, owing to their extensive use in cell death assay. Although DEVE and IETD have been generally considered to be selective for caspase-3 and -8, respectively, the proteolytic cleavage of these substrates does not display absolute specificity for a particular caspase. Thus, we attempted to monitor the cleavage preference for caspase-3, particularly using the recombinant protein substrates. For this aim, the chimeric GST:DEVD:EGFP and GST:IETD:EGFP proteins were genetically constructed by linking GST and EGFP with the linkers harboring DEVD and IETD. To our best knowledge, this work constitutes the first application for the monitoring of cleavage preference employing the recombinant protein substrates that simultaneously allow for mass and fluorescence analyses. Consequently, GST:IETD:EGFP was cleaved partially in response to caspase-3, whereas GST:DEVD:EGFP was completely proteolyzed, indicating that GST:DEVD:EGFP is a better substrate than GST:IETD:EGFP for caspase-3. Collectively, using these chimeric protein substrates, we have successfully evaluated the feasibility of the recombinant protein substrate for applicability to the monitoring of cleavage preference for caspase-3.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.4
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• pp.275-281
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• 2000

Cholecystokinin (CCK) is a gastrointestinal hormone which plays an important role in satiety and gastric motility. It is also widely distributed throughout the central nervous system, where it appears to be involved in the central control of anxiety, feeding behavior and nociception. Two distinct CCK receptor types, $CCK_A$ and $CCK_B,$ have been found in the brain. Both CCK receptors coexist in the rat nucleus tractus solitarius (NTS), which is the primary center for the coordination of peripheral and central activities related to gastrointestinal, cardiovascular and respiratory functions. In order to study ionic actions of CCK on each type of receptor, we investigated the effects of CCK-8S on neurons located in the NTS of the rat using whole-cell patch-clamp recordings in brainstem slices. Application of CCK-8S, under current clamp, produced a membrane depolarization accompanied by action potential firing. This CCK-evoked excitation was dose-dependent $(10\;nM{\sim}10\;{\mu}M)$ and observed in more than 60% of NTS neurons. Under voltage clamp conditions, CCK-8S induced an inward current with a notably increased spontaneous excitatory synaptic activity. However, CCK-8S did not significantly change the amplitude of pharmacologically isolated and evoked EPSP(C)s. Using selective $CCK_A$ and $CCK_B$ receptor antagonists, we observed two different effects of CCK-8S, which suggest $CCK_A$ receptor-mediated inhibitory and $CCK_B$ receptor-mediated excitatory effects in the NTS. These results may help to explain the ability of CCK to modulate gastrointestinal and other reflex systems in the NTS.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.2
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• pp.53-57
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• 2003

The glutamate receptors (GluRs) are key receptors for modulatory synaptic events in the central nervous system. It has been reported that glutamate increases the intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) and induces cytotoxicity. In the present study, we investigated whether the glutamate-induced $[Ca^{2+}]_i$ increase was associated with the activation of ionotropic (iGluR) and metabotropic GluRs (mGluR) in substantia gelatinosa neurons, using spinal cord slice of juvenile rats (10${\sim}21 day). $[Ca^{2+}]_i$ was measured using conventional imaging techniques, which was combined with whole-cell patch clamp recording by incorporating fura-2 in the patch pipette. At physiological concentration of extracellular $Ca^{2+}$, the inward current and $[Ca^{2+}]_i$ increase were induced by membrane depolarization and application of glutamate. Dose-response relationship with glutamate was observed in both $Ca^{2+}$ signal and inward current. The glutamate-induced $[Ca^{2+}]_i$ increase at holding potential of -70 mV was blocked by CNQX, an AMPA receptor blocker, but not by AP-5, a NMDA receptor blocker. The glutamate-induced $[Ca^{2+}]_i$ increase in $Ca^{2+}$ free condition was not affected by iGluR blockers. A selective mGluR (group I) agonist, RS-3,5-dihydroxyphenylglycine (DHPG), induced $[Ca^{2+}]_i$ increase at holding potential of -70 mV in SG neurons. These findings suggest that the glutamate-induced $[Ca^{2+}]_i$ increase is associated with AMPA-sensitive iGluR and group I mGluR in SG neurons of rats.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.6
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• pp.489-495
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• 2014

Protease-activated receptor (PAR)-2 is expressed in endothelial cells and vascular smooth muscle cells. It plays a crucial role in regulating blood pressure via the modulation of peripheral vascular tone. Although some reports have suggested involvement of a neurogenic mechanism in PAR-2-induced hypotension, the accurate mechanism remains to be elucidated. To examine this possibility, we investigated the effect of PAR-2 activation on smooth muscle contraction evoked by electrical field stimulation (EFS) in the superior mesenteric artery. In the present study, PAR-2 agonists suppressed neurogenic contractions evoked by EFS in endothelium-denuded superior mesenteric arterial strips but did not affect contraction elicited by the external application of noradrenaline (NA). However, thrombin, a potent PAR-1 agonist, had no effect on EFS-evoked contraction. Additionally, ${\omega}$-conotoxin GVIA (CgTx), a selective N-type $Ca^{2+}$ channel ($I_{Ca-N}$) blocker, significantly inhibited EFS-evoked contraction, and this blockade almost completely occluded the suppression of EFS-evoked contraction by PAR-2 agonists. Finally, PAR-2 agonists suppressed the EFS-evoked overflow of NA in endothelium-denuded rat superior mesenteric arterial strips and this suppression was nearly completely occluded by ${\omega}$-CgTx. These results suggest that activation of PAR-2 may suppress peripheral sympathetic outflow by modulating activity of $I_{Ca-N}$ which are located in peripheral sympathetic nerve terminals, which results in PAR-2-induced hypotension.

• Journal of Communications and Networks
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• v.11 no.1
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• pp.11-19
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• 2009

Multicarrier direct sequence code division multiple access (MC DS-CDMA) is an attractive technique for achieving high data rate transmission. This is valid regardless of whether or not the potentially large peak-to-average power ratio (PAPR) is an important factor for its application. On the other hand, code select CDMA (CS-CDMA) is an attractive technique with constant amplitude transmission of multicode signal regardless of subchannels. This is achieved by introducing a code select method. In this paper, we propose a new multiple access scheme based on the combination of MC DS-CDMA and CS-CDMA. The proposed scheme, which we call MC CS-CDMA, includes as special cases the subclasses of MC DS-CDMA and CS-CDMA. This paper investigates the performance of these systems over a multipath frequency selective fading channel using a RAKE receiver with maximal ratio combiner. In addition, the PAPR of the proposed system is compared with that of both MC DS-CDMA and CS-CDMA. Simulation results demonstrate that the proposed system provides better PAPR reduction than MC DS-CDMA, at the expense of the complexity of the receiver and the number of available users. The numerical result demonstrates that the proposed system has better performance than MC DS-CDMA due to the increased processing gain and time diversity gain.

• Archives of Pharmacal Research
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• v.20 no.1
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• pp.7-12
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• 1997

Glutamate uptake inhibitor, L-trans-pyrrolidine-2, 4-dicarboxylate (PDC, $20{\mu}M$) elevated basal and N-methyl-D-aspartate (NMDA, $100{\mu}M$)-induced extracellular glutamate accumulation, while it did not augment kainate $100{\mu}M$-induced glutamate accumulation in cultured cerebellar granule neurons. However, pretreatment with PDC for 1 h significantly reduced NMDA-induced glutamate accumulation, but did not affect kainate-induced response. Pretreatment with glutamate $(5{\mu}M)$ for 1 h also reduced NMDA-induced glutamate accumulation, but did not kainate-induced response. Upon a brief application (3-10 min), PDC did neither induce elevation of intracellular calcium concentration $([Ca^{2+}]_i)$ nor modulate NMDA-indLiced $[Ca^{2+}]_1$ elevation. Pretreatment with PDC for 1 h reduced NMDA-induced $[Ca^{2+}]_1$ elevation, but it did not reduce kainate-induced $[Ca^{2+}]_1$ elevation. These results suggest that glutamate concentration in synaptic clefts of neurana cells is increased by prolonged exposure (1 h) of the cells to PDC, and the accumulated glutamate subsequently induces selective desensitization of NMDA receptor.

• The Research Journal of the Costume Culture
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• v.13 no.3 s.56
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• pp.380-390
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• 2005

The effect of high purity chitosan was studied on its application to the natural dyeing using Rhusjara ica. In the dyeing experiment, examinations were introduced on the difference between the chitosan treated fabrics and chitosan untreated fabrics, and on the difference according to the type of mordanting agents and the mordanting methods. Dyeing was carried out for the chitosan treated and untreated fabrics under the conditions of non-mordanting and Al, Sn, Fe mordanting. As a result, we found that, in the case of cotton, chitosan untreated fabrics were not dyed enough, and the dyeing effect was not developed even though the mordanting was introduced. However, the chitosan treated fabrics developed excellent dyeing result even in the non-mordanting case. Through the results of cotton fabric dyeing, it was presumed that the dyeing affinity of the chitosan toward the Rhusjara ica, which prefer specific dyestuff (mainly black color), is selectively high. In the case of nylon, the dyeing effect is not developed easily under all conditions regardless of the chitosan treatment or the mordant treatment. It was inferred that a certain portion of the Rhusjara ica dyestuff made direct bonding with the functional elements in the nylon molecules without the help of the mordant. Through the results of nylon fabrics, it was presumed that some functional elements of the nylon molecules had selective affinity toward the specific colorant (mainly yellow color) of the Rhusjara ica.