• BMB Reports
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• v.45 no.5
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• pp.281-286
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• 2012

Osteoclasts are multinucleated cells that are formed by the fusion of pre-fusion osteoclasts (pOCs). The fusion of pOCs is known to be important for osteoclastic bone resorption. Here, we examined the effect of IFN-${\gamma}$ on the fusion of pOCs. IFN-${\gamma}$ greatly increased the fusion of pOCs in a dose-dependent manner. Furthermore, IFN-${\gamma}$ induced pOC fusion even in hydroxyapatite-coated plates used as a substitute for bone. The resorption area of pOCs stimulated with IFN-${\gamma}$ was significantly higher than that of the control cells. IFN-${\gamma}$ induced the expression of dendritic cell-specific transmembrane protein (DC-STAMP), which is responsible for the fusion of pOCs. IFN-${\gamma}$ enhanced DC-STAMP expression in a dose-dependent manner. The mRNA expression of c-Fos and nuclear factor of activated T cells (NFAT) c1 was enhanced in the pOCs treated with IFN-${\gamma}$. Taken together, these results provide a new insight into the novel role of IFN-${\gamma}$ on the fusion of pOCs.

• IMMUNE NETWORK
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• v.8 no.3
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• pp.98-105
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• 2008

Background: Allergic inflammation was induced by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. A Th2 disproportion was shown in atopic children soon after birth. During specific allergen stimulation, an increase of Th2 cells was observed in most cases. In this study, we prepared new screening "whole blood" system for searching the anti-atopic materials. Cytokine production and IgE secretion from whole blood system were assessed and we confirmed the results by using animal system. Methods: Pathological features in NC/Nga mice are similar to those observed in human atopic dermatitis. Whole blood from NC/Nga mouse was stimulated by using TNCB (Th2 activator) or candidate materials of anti-atopic dermatitis, and the production of cytokines (IL-4, IL-12, and IFN-${\gamma}$) were measured by ELISA. In order to confirm the results of whole blood system, in vivo test was done by using NC/Nga mice. Results: In whole blood system, LPS and extracts of green tea, hardy orange and onion induced the production of IL-12 and IFN-${\gamma}$ while they reduced the production of IL-4. Also, LPS and extracts of onion reduced IgE production. Though atopic dermatitis was observed from a mouse stimulated with TNCB, it was not when a mouse was co-stimulated in LPS or extracts of onion. The results are same as those observed in whole blood system. Conclusion: Whole blood system was simple and speedy methods for searching a materials compared with the conventional high-cost animal system. And the results using whole blood system was proved to be reliable in our experiments for screening anti-atopic material. We expect that the system can be applied to other experiments for searching similar materials.

• BMB Reports
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• v.48 no.7
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• pp.407-412
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• 2015

The 12 kDa FK506-binding protein (FK506BP12), an immunosuppressor, modulates T cell activation via calcineurin inhibition. In this study, we investigated the ability of PEP-1-FK506BP12, consisting of FK506BP12 fused to the protein transduction domain PEP-1 peptide, to suppress catabolic responses in primary human chondrocytes and in a mouse carrageenan-induced paw arthritis model. Western blotting and immunofluorescence analysis showed that PEP-1-FK506BP12 efficiently penetrated chondrocytes and cartilage explants. In interleukin-1β (IL-1β)-treated chondrocytes, PEP-1-FK506BP12 significantly suppressed the expression of catabolic enzymes, including matrix metalloproteinases (MMPs)-1, -3, and -13 in addition to cyclooxygenase-2, at both the mRNA and protein levels, whereas FK506BP12 alone did not. In addition, PEP-1-FK506BP12 decreased IL-1β-induced phosphorylation of the mitogen-activated protein kinase (MAPK) complex (p38, JNK, and ERK) and the inhibitor kappa B alpha. In the mouse model of carrageenan-induced paw arthritis, PEP-1-FK506BP12 suppressed both carrageenan-induced MMP-13 production and paw inflammation. PEP-1-FK506BP12 may have therapeutic potential in the alleviation of OA progression. [BMB Reports 2015; 48(7): 407-412]

• Herbal Formula Science
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• v.27 no.4
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• pp.237-244
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• 2019

Objectives : The purpose of this study was to investigate the effects of Carthami flos on pacemaker potentials of small intestinal and colonic Interstitial Cells of Cajal (ICC). Methods : To dissociate the ICC, we used enzymatic digestions from the small intestine and colon in mice. In the ICC, the electrophysiological whole-cell patch-clamp configuration was used to record pacemaker potentials in the cultured ICC. Results : 1. The ICC generated pacemaker potentials in the murine small intestine and colon. 2. Pretreatment with a Ca²⁺ free solution and thapsigargin, a Ca²⁺-ATPase inhibitor in the endoplasmic reticulum, stopped the pacemaker potentials. In the case of Ca²⁺-free solutions, Carthami flos did not induce membrane depolarizations in the murine small intestine and colon. However, when thapsigargin in a bath solution was applied, Carthami flos induced membrane depolarizations only in the murine colon. 3. Pretreatment with 2-APB (transient receptor potential melastatin (TRPM) channel inhibitor) abolished the pacemaker potentials and suppressed Carthami flos-induced effects in the murine small intestine and colon. 4. However, pretreatment with T16Ainh-AO1 (Ca²⁺ activated Cl- channel; anoctamin 1 (ANO1) inhibitor) did not affect the pacemaker potentials and induced Carthami flos-induced effects only in the murine small intestine. Conclusions : These results suggest that Carthami flos can modulate the pacemaker activity of ICC and the mechanisms underlying pacemaking in ICC might be different in the small intestine and the colon.

• BMB Reports
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• v.41 no.11
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• pp.814-819
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• 2008

Interleukin-32 (IL-32) induces a variety of proinflammatory cytokines and chemokines. The IL-32 transcript was reported originally in activated T cells; subsequently, it was demonstrated to be abundantly expressed in epithelial and endothelial cells upon stimulation with inflammatory cytokines. IL-32 is regulated robustly by other major proinflammatory cytokines, thereby suggesting that IL-32 is crucial to inflammation and immune responses. Recently, an IL-32$\alpha$-affinity column was employed in order to isolate an IL-32 binding protein, neutrophil proteinase 3 (PR3). Proteinase 3 processes a variety of inflammatory cytokines, including TNF$\alpha$, IL-$1{\beta}$, IL-8, and IL-32, thereby enhancing their biological activities. In the current study, we designed four PR3-cleaved IL-32 separate domains, identified by potential PR3 cleavage sites in the IL-32$\alpha$ and $\gamma$ polypeptides. The separate domains of the IL-32 isoforms $\alpha$ and $\gamma$ were more active than the intrinsic $\alpha$ and $\gamma$ isoforms. Interestingly, the N-terminal IL-32 isoform $\gamma$ separate domain evidenced the highest levels of biological activity among the IL-32 separate domains.

• Korean Journal of Clinical Pharmacy
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• v.8 no.1
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• pp.29-34
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• 1998

Murine CD38 is a 42 kDa type II glycoprotein expressed on cell surface of both B and T lymphocytes. CD38 is a multifunctional enzyme that catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD and cADPR hydrolase activity of CD38 catalyzes the hydrolysis of cADPR to ADP-ribose (ADPR). And also, CD38 has the catalytic activity of NAD glycohydrolase (NADase) which catalyzes the hydrolysis of catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD to ADPR. In this study, we attempted to purify CD38 from mouse lymphocytes by using the immobilized anti-CD38 monoclonal antibody. The single step immuno-affinity column chromatography resulted in homogeneous purification, showing a single protein of 42 kDa on a SDS polyacrylamide gel. We have investigated the effects of various inhibitors on the enzyme activities of the purified CD38. Cibacron blue (0.5 mM) inhibited all three enzyme activities of CD38, NADase, ADP-ribosyl cyclase and cADPR hydrolase activities. ADPR (2 mM) showed inhibitory effect on both cADPR hydrolase activity and NADase, but not on ADP-ribosyl cyclase activity. However, ATP (2 mM) inhibited only cADPR hydrolase activity. $Zn^{2+}$ (1 mM) showed similar inhibitory effect as that of ADPR, but activated cyclase activity These results suggest that CD38 has three different catalytic activity domains which might be differentially regulated by their specific inhibitors.

• Genomics & Informatics
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• v.4 no.2
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• pp.71-76
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• 2006

We have investigated biological responses to radiofrequency (RF) radiation in in vitro and in vivo models. By measuring the levels of heat shock proteins as well as the activation of mitogen activated protein kinases (MAPKs), we could not detect any differences upon RF exposure. In this study, we used more sensitive method to find the molecular responses to RF radiation. Jurkat, human T-Iymphocyte cells were exposed to 1763 MHz RF radiation at an average specific absorption rate (SAR) of 10 W/kg for one hour and harvested immediately (R0) or after five hours (R5). From the profiles of 30,000 genes, we selected 68 differentially expressed genes among sham (S), R0 and R5 groups using a random-variance F-test. Especially 45 annotated genes were related to metabolism, apoptosis or transcription regulation. Based on support vector machine (SVM) algorithm, we designed prediction model using 68 genes to discriminate three groups. Our prediction model could predict the target class of 19 among 20 examples exactly (95% accuracy). From these data, we could select the 68 biomarkers to predict the RF radiation exposure with high accuracy, which might need to be validated in in vivo models.

• Proceedings of the Korean Vacuum Society Conference
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• 2010.02a
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• pp.322-322
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• 2010

For boron doping on n-type silicon wafer, around $1,000^{\circ}C$ doping temperature is required, because of the relatively low solubility of boron in a crystalline silicon comparing to the phosphorus case. Boron doping by fiber laser annealing and lamp furnace heat treatment were carried out for the uniformly deposited p-a-Si:H layer. Since the uniformly deposited p-a-Si:H layer by cluster is highly needed to be doped with high temperature heat treatment. Amorphous silicon layer absorption range for fiber laser did not match well to be directly annealed. To improve the annealing effect, we introduce additional lamp furnace heat treatment. For p-a-Si:H layer with the ratio of $SiH_4:B_2H_6:H_2$=30:30:120, at $200^{\circ}C$, 50 W power, 0.2 Torr for 30 min. $20\;mm\;{\times}\;20\;mm$ size fiber laser cut wafers were activated by Q-switched fiber laser (1,064 nm) with different sets of power levels and periods, and for the lamp furnace annealing, $980^{\circ}C$ for 30 min heat treatment were implemented. To make the sheet resistance expectable and uniform as important processes for the $p^+$ layer on a polished n-type silicon wafer of (100) plane, the Q-switched fiber laser used. In consequence of comparing the results of lifetime measurement and sheet resistance relation, the fiber laser treatment showed the trade-offs between the lifetime and the sheet resistance as $100\;{\omega}/sq.$ and $11.8\;{\mu}s$ vs. $17\;{\omega}/sq.$ and $8.2\;{\mu}s$. Diode level device was made to confirm the electrical properties of these experimental results by measuring C-V(-F), I-V(-T) characteristics. Uniform and expectable boron heavy doped layers by fiber laser and lamp furnace are not only basic and essential conditions for the n-type crystalline silicon solar cell fabrication processes, but also the controllable doping concentration and depth can be established according to the deposition conditions of layers.

• Biomolecules & Therapeutics
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• v.8 no.3
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• pp.276-280
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• 2000

In order to identify the domains of the G$_{\alpha}$16 subunit G protein that are responsible for its activation by the Interleukin-8 receptor, a serious of chimeras between G$_{\alpha}$16 and G$_{\alpha}$11 were assessed for their abilities to be activated by these receptors. Co-expression of IL-8 receptor and chimeras in which the carboxyl-terminal regions of G$_{\alpha}$11 were replaced from 30 up to 156 amino acid residues with the corresponding regions of G$_{\alpha}$16 demonstrated that C-terminal 156 amino acid residues of the G$_{\alpha}$16 were not sufficient to confer IL-8 receptor interaction specificity. Testing of a reciprocal serious of chimeras composed of G$_{\alpha}$16 sequences at the amino terminus and G$_{\alpha}$11 sequences at the carboxyl terminals revealed that sequences extending from the amino tar- minus to amino acid 209 of G$_{\alpha}$16 were sufficient to 7ndow the chimera with 75-80% of interaction specificity for 7-8-induced activation. These results suggest th,.7t combined interactions of the C-terminal 30 amino acid residues and certain domains extending from the arts.ino terminus to amino acid 209 of Gal 6 protein may be involved in its couplings to IL-8 receptor.tain domains extending from the arts.ino terminus to amino acid 209 of Gal 6 protein may be involved in its couplings to IL-8 receptor.

• Journal of Microbiology and Biotechnology
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• v.32 no.7
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• pp.835-843
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• 2022

Deinococcus radiodurans is an extremophilic bacterium that can thrive in harsh environments. This property can be attributed to its unique metabolites that possess strong antioxidants and other pharmacological properties. To determine the potential of D. radiodurans R1 lysate (DeinoLys) as a pharmacological candidate for inflammatory bowel disease (IBD), we investigated the antiinflammatory activity of DeinoLys in bone marrow-derived dendritic cells (BMDCs) and a colitis mice model. Lipopolysaccharide (LPS)-stimulated BMDCs treated with DeinoLys exhibited alterations in their phenotypic and functional properties by changing into tolerogenic DCs, including strongly inhibited proinflammatory cytokines (TNF-α and IL-12p70) and surface molecule expression and activated DC-induced T cell proliferation/activation with high IL-10 production. These phenotypic and functional changes in BMDCs induced by DeinoLys in the presence of LPS were abrogated by IL-10 neutralization. Furthermore, oral administration of DeinoLys significantly reduced clinical symptoms against dextran sulfate sodium-induced colitis, including body weight loss, disease activity index, histological severity in colon tissue, and lower myeloperoxidase level in mice. Our results establish DeinoLys as a potential anti-inflammatory candidate for IBD therapy.