• Molecules and Cells
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• v.40 no.9
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• pp.667-676
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• 2017

Abnormal differentiation of muscle is closely associated with aging (sarcopenia) and diseases such as cancer and type II diabetes. Thus, understanding the mechanisms that regulate muscle differentiation will be useful in the treatment and prevention of these conditions. Protein lysine acetylation and methylation are major post-translational modification mechanisms that regulate key cellular processes. In this study, to elucidate the relationship between myogenic differentiation and protein lysine acetylation/methylation, we performed a PCR array of enzymes related to protein lysine acetylation/methylation during C2C12 myoblast differentiation. Our results indicated that the expression pattern of HDAC11 was substantially increased during myoblast differentiation. Furthermore, ectopic expression of HDAC11 completely inhibited myoblast differentiation, concomitant with reduced expression of key myogenic transcription factors. However, the catalytically inactive mutant of HDAC11 (H142/143A) did not impede myoblast differentiation. In addition, wild-type HDAC11, but not the inactive HDAC11 mutant, suppressed MyoD-induced promoter activities of MEF2C and MYOG (Myogenin), and reduced histone acetylation near the E-boxes, the MyoD binding site, of the MEF2C and MYOG promoters. Collectively, our results indicate that HDAC11 would suppress myoblast differentiation via regulation of MyoD-dependent transcription. These findings suggest that HDAC11 is a novel critical target for controlling myoblast differentiation.

• The Journal of Pediatrics of Korean Medicine
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• v.28 no.4
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• pp.1-29
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• 2014

Objectives Dictamni Radicis Cortex extracts (DRC) has been known to suppress allergic reaction, however the cellular target of DRC and its mode of action remain unclear. The purpose of this study is to investigate the effects of Dictamni Radicis Cortex extracts on DNCB induced atopic dermatitis-like skin lesions of NC/Nga mouse. Methods This study was designed to investigate the effects of DRC extract in the DNP-IgE-induced activation of MC/9 murine mast cell lines in vitro and in the DNCB-induced activation of NC/Nga mouse in vivo. For this investigation, We examined IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF mRNA expression by Real-time PCR, IL-13, MIP-$1{\alpha}$ production by ELISA analysis and manifestations of NFAT1, NFAT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting in vitro. Then, we examined WBC, eosinophil and neutrophil in NC/Nga mouse, IL-5, IL-13 in serum, IFN-${\gamma}$, IL-4 in the spleenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $^+Gr-1^+CD11b$, $B220^+CD23^+$ in the ALN, PBMCs and dorsal skin, IL-5, IL-13 in the dorsal skin by Real-time PCR and the distribution of mast cells by H&E and toluidine blue. Results In vitro the mRNA expression of IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$, GM-CSF and IL-13, MIP-$1{\alpha}$ production by ELISA analysis were completely abolished by DRC and the western blot analysis decreased the expression of mast cell-specific transcription factors including NFAT-1, NF-${\kappa}B$ p65. In vivo DRC oral adminstration also decreased the counts of WBC, eosinophils and inflammatory cytokines such as IL-13 and IgE in the serum. DRC oral adminstration elevated IL-4 level in the spleenocyte culture supernatant. DRC oral adminstration decreased total ALN cells, total skin cells, cell numbers of $CD4^+$, $B220^+CD23^+$ in the ALN, $^+Gr-1^+CD11b$ in the PBMCs and $CD4^+$, $CD8^+$ in the dorsal skin. The mRNA expression of IL-5, IL-13, thickness of epidermis, inflammation immune cells and mast cells were abolished by DRC in the dorsal skin. Conclusions Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mouse were much improved by DRC oral adminstration. These results, therefore, suggest that DRC can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis induced in NC/Nga mouse, and may play an important role in recovering AD symptoms.

• The Journal of Pediatrics of Korean Medicine
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• v.30 no.4
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• pp.29-59
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• 2016

Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

• Parasites, Hosts and Diseases
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• v.54 no.6
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• pp.759-768
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• 2016

Cystic echinococcosis (CE) treatment urgently requires a novel drug. The p38 mitogen-activated protein kinases (MAPKs) are a family of Ser/Thr protein kinases, but still have to be characterized in Echinococcus granulosus. We identified a 1,107 bp cDNA encoding a 368 amino acid MAPK protein (Egp38) in E. granulosus. Egp38 exhibits 2 distinguishing features of p38-like kinases: a highly conserved T-X-Y motif and an activation loop segment. Structural homology modeling indicated a conserved structure among Egp38, EmMPK2, and H. sapiens $p38{\alpha}$, implying a common binding mechanism for the ligand domain and downstream signal transduction processing similar to that described for $p38{\alpha}$. Egp38 and its phosphorylated form are expressed in the E. granulosus larval stages vesicle and protoscolices during intermediate host infection of an intermediate host. Treatment of in vitro cultivated protoscolices with the p38-MAPK inhibitor ML3403 effectively suppressed Egp38 activity and led to significant protoscolices death within 5 days. Treatment of in vitro-cultivated protoscolices with $TGF-{\beta}1$ effectively induced Egp38 phosphorylation. In summary, the MAPK, Egp38, was identified in E. granulosus, as an anti-CE drug target and participates in the interplay between the host and E. granulosus via human $TGF-{\beta}1$.

• Journal of Life Science
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• v.30 no.5
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• pp.476-482
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• 2020

In rice agriculture, Eleocharis kuroguwai Ohwi (olbanggae) is a target for herbicidal intervention as a problem weed although it has also long been used clinically as a traditional medicine for jaundice, fever, and blood flow. E. kuroguwai has been evaluated in many clinical trials, but its molecular biological advantages are still unknown. Here, we investigate the anti-inflammatory effects of E. kuroguwai 80% ethanol extracts by screening NO production in LPS-induced macrophage activation. To find the most effective fractions, we partitioned five sub-fractions using HP20 column chromatography, namely 20%, 40%, 60%, 80%, and 100%. Of these, the 60% and 80% sub-fractions were found to significantly inhibit NO production; there were no toxicological effects at any concentration. In addition, the 80% sub-fraction inhibited significantly the iNOS and the mRNA of the pro-inflammatory mediators IL-6, TNF-α, and IL-1β by inhibiting the phosphorylation of JNK and ERK pathways associated with MAPKs signaling. Our results suggest that the 80% E. kuroguwai sub-fraction has the most significant anti-inflammatory effects of inhibiting iNOS and pro-inflammatory mediators and suppressing the phosphorylation of JNK and ERK. Therefore, the 80% sub-fraction of E. kuroguwai extract may be a therapeutic candidate for inflammatory diseases associated with the overexpression of MAPKs.

• Proceedings of the Korean Society of Medical Physics Conference
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• 2002.09a
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• pp.400-403
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• 2002

The fundamental experiments for measuring soft x-ray characteristics from the vacuum capillary are described. These experiments were primarily performed in order to generate line spectra such as x-ray lasers. The generator consists of a high-voltage power supply, a polarity-inversion ignitron pulse generator, a turbo-molecular pump, and a radiation tube with a capillary. A high-voltage condenser of 200 nF in the pulse generator is charged up to 20 kV by the power supply, and the electric charges in the condenser are discharged to the capillary in the tube after closing the ignitron. During the discharge, weakly ionized plasma forms on the inner and outer sides of a capillary. In the present work, the pump evacuates air from the tube with a pressure of about 1 mPa, and a demountable capillary was developed in order to measure x-ray spectra according to changes in the capillary length. In this capillary, the anode (target) and cathode elements can be changed corresponding to the objectives. The capillary diameter is 2.0 mm, and the length is adjusted from 1 to 50 mm. When a capillary with aluminum anode and cathode electrodes was employed, both the cathode voltage and the discharge current almost displayed damped oscillations. The peak values of the voltage and current increased when the charging voltage was increased, and their maximum values were -10.8 kV and 4.7 kA, respectively. The x-ray durations observed by a 1.6 ${\mu}$m aluminum filter were less than 30 ${\mu}$s, and we detected the aluminum characteristic x-ray intensity using a 6.8 ${\mu}$m aluminum filter. In the spectrum measurement, two sets of aluminum and titanium electrodes were employed, and we observed multi-line spectra. The line photon energies seldom varied according to changes in the condenser charging voltage and to changes in the electrode element. In the case where the titanium electrode was employed, the line number decreased with corresponding decreases in the capillary length. Compared with incoherent visible light, these rays from the capillary were diffracted and diffused greatly after passing through two slits.

• Korean journal of applied entomology
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• v.52 no.1
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• pp.35-43
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• 2013

All tested Korean populations of the diamondback moth, Plutella xylostella, are known to be resistant especially against pyrethroid insecticides by mutation in its molecular target, para-sodium channel. Moreover, P. xylostella is able to develop resistance against most commercial insecticides. This study was performed to develop an efficient control technique against P. xylostella by a combined treatment of an endoparasitoid wasp, Cotesia plutellae, and a microbial insecticide, Bacillus thuringiensis. To investigate any parasitism preference of C. plutellae against susceptible and resistant P. xylostella, five different populations of P. xylostella were compared in insecticide susceptibilities and parasitism by C. plutellae. These five P. xylostella populations showed a significant variation against three commercial insecticides including pyrethroid, organophosphate, neonicotinoid, and insect growth regulator. However, there were no significant differences among five P. xylostella populations in their parasitic rates by C. plutellae. Moreover, parasitized larvae of P. xylostella showed significantly higher susceptibility to B. thuringiensis. As an immunosuppressive agent, viral ankyrin genes (vankyrins) encoded in C. plutellae were transiently expressed in nonparasitized larvae. Expression of vankyrins significantly enhanced the efficacy of B. thuringiensis against the third instar larvae of P. xylostella. Thus an immunosuppression induced by C. plutellae enhanced the insecticidal efficacy of B. thuringiensis. These results suggest that a combined treatment of C. plutellae and B. thuringiensis may effectively control the insecticide-resistant populations of P. xylostella.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.05a
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• pp.14-14
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• 2010

Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

• Korean Journal of Plant Resources
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• v.33 no.4
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• pp.271-278
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• 2020

In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of the Berchemia floribunda (BF) which belongs to Rhamnaceae against human colorectal cancer cells. The treatment of BF decreased the cell proliferation in HCT116 cell and suppressed cellular accumulation of Cyclin D1 protein. Inhibition of proteasomal activity by MG132 attenuated BF-mediated Cyclin D1 downregulation and Cyclin D1 was decreased in the cell treated with BF. These findings indicates that BF-mediated Cyclin D1 downregulation may be result from Cyclin D1 proteasomal degradation. Additionally, BF-mediated Cyclin D1 degradation was blocked in the presence of LiCl, a GSK3β inhibitor, but not PD98059, SP600125, SB203580, Bay11-7082, LY294002 an ERK1/2 inhibitor, JNK inhibitor, p38 inhibitor, IκK inhibitor and PI3K inhibitor. Furthermore, BF phosphorylated Cyclin D1 at threonine-286 (Thr286), and LiCl-induced GSK3β inhibition reduced the BF mediated phosphorylation of Cyclin D1 at Thr286. These results suggested that BF may downregulate Cyclin D1 expression as a potential anti-cancer target through GSK3β dependent Cyclin D1 degradation. Therefore, this study provides that the extract of BF has anticancer activity against human colorectal cancer cells.

• Journal of the Korean Society for Nondestructive Testing
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• v.35 no.2
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• pp.112-119
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• 2015

Photoacoustic microscopy is capable of providing high-resolution molecular images, and its spatial resolution is typically determined by ultrasonic transducers used to receive the photoacoustic signals. Therefore, ultrasonic transducers for photoacoustic microscopy (PAM) should have a high operating frequency, broad bandwidth, and high signal-reception efficiency. Polyvinylidene fluoride (PVDF) is a suitable material. To take full advantage of this material, the selection of the backing material is crucial, as it influences the center frequency and bandwidth of the transducer. Therefore, we experimentally determined the most suitable backing material among EPO-TEK 301, E-Solder 3022, and RTV. For this, three PVDF high-frequency single-element transducers were fabricated with each backing material. The center frequency and -6 dB bandwidth of each transducer were ascertained by a pulse-echo test. The spatial resolution of each transducer was examined using wire-target images. The experimental results indicated that EPO-TEK 301 is the most suitable backing material for a PAM transducer. This material provides the highest signal magnitude and a reasonable bandwidth because a large portion of the energy propagates toward the front medium, and the PVDF resonates in the half-wave mode.