• Journal of Life Science
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• v.25 no.2
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• pp.121-129
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• 2015

Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.

• Molecules and Cells
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• v.20 no.3
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• pp.385-391
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• 2005

As a first step towards identifying genes involving in the signal transduction pathways mediating rice blast resistance, we isolated 3 mutants lines that showed enhanced susceptibility to rice blast KJ105 (91-033) from a T-DNA insertion library of the japonica rice cultivar, Hwayeong. Since none of the susceptible phenotypes co-segregated with the T-DNA insertion we adapted a map-based cloning strategy to isolate the gene(s) responsible for the enhanced susceptibility of the Hwayeong mutants. A genetic mapping population was produced by crossing the resistant wild type Hwayeong with the susceptible cultivar, Nagdong. Chi-square analysis of the $F_2$ segregating population indicated that resistance in Hwayeong was controlled by a single major gene that we tentatively named Pi-hy. Randomly selected susceptible plants in the $F_2$ population were used to build an initial map of Pi-hy. The SSLP marker RM2265 on chromosome 2 was closely linked to resistance. High resolution mapping using 105 $F_2$ plants revealed that the resistance gene was tightly linked, or identical, to Pib, a resistance gene with a nucleotide binding sequence and leucine-rich repeats (NB-LRR) previously isolated. Sequence analysis of the Pib locus amplified from three susceptible mutants revealed lesions within this gene, demonstrating that the Pi-hy gene is Pib. The Pib mutations in 1D-22-10-13, 1D-54-16-8, and 1C-143-16-1 were, respectively, a missense mutation in the conserved NB domain 3, a nonsense mutation in the 5th LRR, and a nonsense mutation in the C terminus following the LRRs that causes a small deletion of the C terminus. These findings provide evidence that NB domain 3 and the C terminus are required for full activity of the plant R gene. They also suggest that alterations of the resistance gene can cause major differences in pathogen specificity by affecting interactions with an avirulence factor.

• Molecules and Cells
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• v.19 no.3
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• pp.402-407
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• 2005

Bone marrow mesenchymal stem cells (MSCs) have shown potential for cardiac repair following myocardial injury, but this approach is limited by their poor viability after transplantation. To reduce cell loss after transplantation, we introduced the fibroblast growth factor-2 (FGF-2) gene ex vivo before transplantation. The isolated MSCs produced colonies with a fibroblast-like morphology in 2 weeks; over 95% expressed CD71, and 28% expressed the cardiomyocyte-specific transcription factor, Nkx2.5, as well as ${\alpha}$-skeletal actin, Nkx2.5, and GATA4. In hypoxic culture, the FGF-2-transfected MSCs (FGF-2-MSCs) secreted increased levels of FGF-2 and displayed a threefold increase in viability, as well as increased expression of the anti-apoptotic gene, Bcl2, and reduced DNA laddering. They had functional adrenergic receptors, like cardiomyocytes, and exposure to norepinephrine led to phosphorylation of ERK1/2. Viable cells persisted 4 weeks after implantation of $5.0{\times}10^5$ FGF-2-MSCs into infarcted myocardia. Expression of cardiac troponin T (CTn T) and a voltage-gated $Ca^{2+}$ channel (CaV2.1) increased, and new blood vessels formed. These data suggest that genetic modification of MSCs before transplantation could be useful for treating myocardial infarction and end-stage cardiac failure.

• Applied Biological Chemistry
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• v.41 no.5
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• pp.390-394
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• 1998

The tuber of Liriope platyphylla was extracted with 80% aqueous MeOH and solvent-fractionated with EtOAc, n-BuOH and $H_2O$. Evaluation of growth inhibitory activity of each fractions on various carcinoma cells, A549, SK-OV-3, SK-Mel-2, XF-498 and HCT-15, indicated the n-BuOH fraction to be the highest in the activity. From the fraction, two saponin compounds were isolated through Amberlite XAD-II and silica gel column chromatographies, repeatedly, and their chemical structures were elucidated as spicatoside A and B by interpretation of spectral data, NMR and IR, and adaptation of acid hydrolysis. Spicatoside A showed growth inhibitory activity on carcinoma cells, and the $IC_{50}$ values against A549, SK-OV-3, SK-Mel-2, XF-498 and HCT-15 cells were determined to be 17.3, 21.7, 14.9, 18.8 and $15.6\;{\mu}g/ml$, respectively.

• Restorative Dentistry and Endodontics
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• v.30 no.5
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• pp.402-408
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• 2005

The purpose of this study was to investigate the effects of estrogen deficiency on pulpodentinal complex of tooth in ovariectomized rats. Thirty female Sprague-Dawley rats, 10 weeks old, were used. Rats were grouped into two groups. One group (n = 15) was subjected to sham surgery (SHAM) and the other group (n = 15) was ovariectomized bilaterally (OVX). Animals were sacrificed 12 weeks later, and their mandibular molars and associated periodontal supporting tissues were dissected out, and fixed in $10\%$ buffered formalin. For comparison of groups, immunostained for osteonectin. Histomorphometrical measurement of change of teeth was performed using an image analysis system and paired t-test was used and the level of significance for overall differences was set at p < 0.05. In immunostaining of osteonectin, they were significantly different from each other. The predentin thickness in OVX rats was wider than in SHAM rats. And in SHAM rats, osteonectin was more specifically stained in predentin areas than in OVX rats. These results indicate that estrogen deficiency increased the unmineralized predentin areas and decreased osteonectin content in pulpal tissues in rats. If our result is applicable to human studies, odotoblast is affected by estrogen deficiency.

• Journal of Gastric Cancer
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• v.5 no.1
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• pp.52-56
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• 2005

Despite improvements in the surgical treatment of gastric adenocarcinomas, the recurrence rates remain high in patients with advanced-stage disease. Most of the recurrence occurs within 3 years of the surgical resection, and nearly $90\%$ of the patients with recurrence die within 2 years of the diagnosis of recurrence. A recent study analyzed recurrence patterns for patients who had undergone a potentially curative gastrectomy. For those patients, $33\%$ of the recurrences involved locoregional sites, $44\%$ the peritoneum, and $38\%$ distant sites. A 51-year-old female patient was diagnosed with stomach cancer and underwent a total gastrectomy with D2 lymph node dissection during Oct. 1999. The pathologic report indicated a T3N1M0 tumor. We performed immunochemotherapy for 2 years with regular follow up. A gastrofiberscopic examination done during sep. 2004, cancer recurrence was found at the Kim's tie site of the jejunual loop. We did an abdominal exploration and a segmental resection of cancer site with pathologically negative resection margins. After the operation, we started secondary chemotherapy with TS-1.

• KSBB Journal
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• v.30 no.4
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• pp.175-181
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• 2015

Cells proliferate via repeating process that growth and division. This process is G1, S, G2 and M four phases consists. Monitoring the progression of the cell cycle is a specific step that to be a continuous process is repeated to adjust the start of the next step. At this time, this process is called a Checkpoint. Currently, there are three known checkpoints that G1-S phase, G2-M phase, and the M phase. In this study, we confirmed that cell cycle arrest effects by ethanol extracts of Artemisia annua Linne (AAE) in Hep3B liver cancer cells. AAE was regulated proteins which involved in cell cycle such as pAkt, pMDM2, p53, p21, pCDK2 (T14/Y15). AAE induced cell cycle arrest in G1 checkpoint through phosphorylation of CDK2. Akt and p53 upstream is inhibited by AAE and p53 activated by non-activated pMDM2, p53 inhibitor. Thereby, activated p53 is transcript to p21 and activated p21 protein is combined with Cyclin E-pCDK2 complex. Therefore, we confirmed that AAE-induced cell cycle arrest was occurred by p21-Cyclin E-pCDK2 complex by inhibition of pAkt signal. Because of this cell cycle can't pass to S phase from G1 phase.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.44 no.2
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• pp.191-200
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• 2018

In this study, we conducted research to find anti-wrinkle skin care ingredients from natural sources via the measurement of procollagen type 1 levels in the medium of normal human dermal fibroblast (NHDF) and found that a Nelumbo nucifera leave extract (NLE) was as the best effective ingredient. By the high performance liquid chromatography analysis, we confirmed isoquercitrin was one of the main compounds of NLE. Moreover, it also increased the procollagen type 1 production without cytotoxicity of NHDF. NLE and isoquercitrin exerted free radical scavenging activity. Especially, isoquercitrin exhibited strong intracellular antioxidant capacity in human skin-derived cells, HaCaT and NHDF. Finally, we performed clinical test for the inhibitory effect of NLE on skin wrinkle formation. A randomized study was conducted in 22 healthy female volunteers, aged between 30 and 65 yrs, with moderate to moderately severe facial wrinkles. Volunteer applied a 1.7% NLE cream (isoquercitrin 51 ppm) and a control cream at each facial side (left/right) twice daily for 8 weeks. The 1.7% NLE cream improved skin roughness through reducing the wrinkle of the craw's feet significantly without any skin side effect. Our results demonstrate that NLE and isoquercitrin can increase the collagen production and exert antioxidant activity. Therefore, we expect that the new non-toxic herbal extract, isoquercitrin-containing NLE will be interesting natural ingredient of cosmetics with anti-wrinkle efficacy.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.1 s.32
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• pp.66-79
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• 2007

Objectives : This study was carried out to evaluate anti-tummor effect about apoptosis of Cheongyulsodok-Eum (CSE) Results : 1. Anti-tumor(HL-60 cells) effects of CSE water extracts(Exts) were more effective in high density.($IC_{50:}:572$ ${\mu}g/ml$) 2. The generation of $O_2\;^-$ in HL-60 cells were according to the concentration of CSE water Exts, specially more effective on 100 ${\mu}g/ml$ and 1000 ${\mu}g/ml$ concentration. 3. The SOD activities in HL-60 cells were in proportion as cytotoxicity against HL-60 cells of CSE water Exts. 4. The GPx activities in HL-60 cells were in proportion as cytotoxicity against HL-60 cells of CSE water Exts(more effective on 100 ${\mu}g/ml$ and 1000 ${\mu}g/ml$ concentration), but the catalase activities in HL-60 cells were not effective. 5. DPPH radical scavenging activity of CSE water Exts was effective.(3 ${\mu}g/ml:31.2{\pm}5.2$ %, 10 ${\mu}g/ml:49.6{\pm}7.3$ %, 30 ${\mu}g/ml:35.8{\pm}5.7$ % 100 ${\mu}g/ml:42.3{\pm}6.4$ %) 6. The results of cytotoxicity against HL-60 cells of CSE were as follows. 1) In hexane fraction, the cytotoxicity against HL-60 cells($IC_{50:}:592$ ${\mu}g/ml$) was more effective than against NIH3T3 cells. 2) In ethyl acetate fraction, the cytotoxicity against HL-60 cells was not effective. 3) In butanol fraction, the cytotoxicity against HL-60 cell($IC_{50:}:306$ ${\mu}g/ml$) was more effective than against NIH3T3 cells. 4) In $H_2O$ fraction, the cytotoxicity against HL-60 cells was not effective. Conclusion : These result suggest that CSE has antioxidative effects and anti-tumor effects by apoptosis of free radical($O_2\;^-$) activity, especially butanol and hexane fraction from water extract has more effective in anti-twnor effects.

• Korean Journal of Plant Resources
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• v.16 no.3
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• pp.230-237
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• 2003

This study was conducted to investigate the occurrence and characterization of tobacco mosaic virus(TMV) in Chinese foxglove isolated from the field of the Chonbuk province(Jinan, Jangsu, Jeongeup). TMV was detected in all three regions and confirmed positive reaction by ELISA test. In the host range test, Chenopodium amaranticola, Nicotiana glutinosa, N. tabacum cv. 'Bright yellow', N. tabacum cv. 'KY­57, Datura stramonium were locally infected with the virus. The virus produced mosaic symptom on inoculated leaves of N. tabacum cv. 'Samson'. However, Chenopodium quinoa, Glycine max, Raphanus sativus, Cucumis sativus, Cucurbita moschata, Brassica rape and Lycopersion esculentum did not show any symptoms. TMV particles were revealed as a stiff rod shape by transmission electron microscopic(TEM) and measured as 300 nm in length with 18 nm in diameter. Total RNA was extracted from showing symptom loaves infected with TMV and the reverse transcription­polymerase chain reaction (RT­PCR) obtained 531 bp DNA product of RNA with specific primer used. The capsid protein of TMV­RE showed higher amino acid sequence homology(97.7％) with TMV­To than with TMV­P(72.2％). The capsid protein of TMV­152 showed same amino acid sequence homology with TMV­F. The result of comparison of nucleotides sequence homology between TMV­RE strain and other TMV strain showed 94％ homology with others except TMV­P(67.3％) and TMV ­ C(68.6％).