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Isolation and characterization of human dental tissue-derived stem cells in the impacted wisdom teeth: comparison of dental follicle, dental pulp, and root apical papilla-derived cells (미성숙 매복지치의 치낭, 치수, 치근유두 조직에서 다능성 줄기세포의 분리와 특성화에 대한 연구)

  • Song, Jung-Ho;Park, Bong-Wook;Byun, June-Ho;Kang, Eun-Ju;Rho, Gyu-Jin;Shin, Sang-Hun;Kim, Uk-Kyu;Kim, Jong-Ryoul
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.3
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    • pp.186-196
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    • 2010
  • Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

Water Transport Characteristics of Paddy Plow Pan Soils as Estimated by Particle Size Distribution Fractal Dimension (토양입자분포 프랙탈차원을 활용한 논토양 쟁기바닥층 물이동 추정)

  • Han, Kyung-Hwa;Cho, Hyun-Jun;Hur, Seung-Oh;Ha, Sang-Geun;Cho, Hee-Rae;Jeon, Sang-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.1
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    • pp.1-7
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    • 2010
  • This study was carried out to investigate plow pan characteristics and to grasp the relationship between its particle size distribution fractal dimension ($D_m$) and water transport in paddy plow pan. Twenty four soil sampling sites with different management groups, ordinary and sandy-textured, were selected and investigated for physical properties of soils such as Yamanaka hardness in April, non-submerged condition, before rice seedling transplanting. The plow pan appearing depth and thickness was determined by penetration resistance profile. Undisturbed core samples with five replicates were sampled at plow pan layerwith 2 inch cores for measuring soil bulk density and saturated hydraulic conductivity. The particle size distribution fractal dimension ($D_m$) was calculated by the method following the procedure Tylerand Wheacraft (1992), using the USDA-based particle size analysis datawith fractions of 0-0.002, 0.002-0.053, 0.053-0.1, 0.1-0.25, 0.25-0.5, 0.5-1.0, and 1.0-2.0 mm. The plow pan of investigated fields appeared at a range from 5 to 30 cm depth, showing minimum value in sandy-textured management group and maximum value in ordinary management group. The thickness of plow pan were distributed from 5 to 17 cm, showing both minimum and maximum values in sandy-textured management group. Averagely, the plow appearing depth were deeper in ordinary management group than in sandy-textured management group, whereas the reverse in the thickness of plow pan. The particle size distribution fractal dimension ($D_m$) had higher value with finer textures, with higher fractality in coarser texture. Saturated hydraulic conductivities, $K_s$, of plow pan soils distributed from 0.5 to 1420 mm $day^{-1}$, having the highest value in sandy skeletal soils. The $K_s$ decreased with decreasing clay content and $D_m$, showing power function relationships. The coefficient of determination, $R^2$, of the fitted power functions were higher in $D_m$ as x-axis than in clay content. This means that $D_m$ could give us more effective estimation than clay content. Especially, sandy-textured paddy soils had higher $R^2$, compared to ordinary paddy soils. $K_s$ of relatively coarse-textured soils with less than 18%of clay content, therefore, was more dependent on particle size distribution than that of relatively fine-textured soils. From these results, it could be concluded that the fractal scaling gives us a unique quantity describing particle size distribution and then can be applied to estimate saturated hydraulic conductivity, especially more effective in coarse-textured soils.

Processed Panax ginseng, sun ginseng, inhibits the differentiation and proliferation of 3T3-L1 preadipocytes and fat accumulation in Caenorhabditis elegans

  • Lee, Hyejin;Kim, Jinhee;Park, Jun Yeon;Kang, Ki Sung;Park, Joeng Hill;Hwang, Gwi Seo
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.257-267
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    • 2017
  • Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.

Improving Corsican pine somatic embryo maturation: comparison of somatic and zygotic embryo morphology and germination

  • Wtpsk, Senarath;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.61-62
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    • 2003
  • Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

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An Empirical Study on the Long-Run Performance of Cross-Listings by Multinational Corporations (다국적기업 해외상장의 장기적인 성과에 관한 연구)

  • Kim, Dong-Soon;Park, Sang-An
    • The Korean Journal of Financial Management
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    • v.21 no.2
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    • pp.27-63
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    • 2004
  • Since the 1980s, many multinational corporations have been issuing stocks on foreign stock exchanges, not only to enhance their investor base and liquidity, but also to diversify risks. The phenomenon has also been intensified by the rapid financial globalization and securitization trends. The main purpose of this study is to look into the long-run performance of MNCs' cross-listings of stocks on foreign stock exchanges. We use the event study and cross-sectional regression methods. We obtained some interesting empirical results about the long-run effect of cross-listings. First before the listing data the effect of cross-listing is to increase the underlying stock Vice in the local market. It may be caused by expectation of lower risk and cost of capital. However, after the listing data the stock price has been declining, even if it is not significant. Second, we examine the difference in the long-run cross-listing effect, which may be caused by the listing direction. When listing is made from a less developed market to a more developed market, the effect is better than that in the reverse direction. Furthermore, the effect is worse, when the listing company's home country is the U.S. Third, there is a negative relation between CARs and underlying stock liquidity in the local market, So it implies that a firm, whose underlying stocks are very liquid in the local market should carefully value cross-listing based upon the cost and benefit analysis. Last, but not the least we find that the long-un cross-listing effect is better, when a listing firm's ROE is higher.

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Discounted Cost Model of Condition-Based Maintenance Regarding Cumulative Damage of Armor Units of Rubble-Mound Breakwaters as a Discrete-Time Stochastic Process (경사제 피복재의 누적피해를 이산시간 확률과정으로 고려한 조건기반 유지관리의 할인비용모형)

  • Lee, Cheol-Eung;Park, Dong-Heon
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.29 no.2
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    • pp.109-120
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    • 2017
  • A discounted cost model for preventive maintenance of armor units of rubble-mound breakwaters is mathematically derived by combining the deterioration model based on a discrete-time stochastic process of shock occurrence with the cost model of renewal process together. The discounted cost model of condition-based maintenance proposed in this paper can take into account the nonlinearity of cumulative damage process as well as the discounting effect of cost. By comparing the present results with the previous other results, the verification is carried out satisfactorily. In addition, it is known from the sensitivity analysis on variables related to the model that the more often preventive maintenance should be implemented, the more crucial the level of importance of system is. However, the tendency is shown in reverse as the interest rate is increased. Meanwhile, the present model has been applied to the armor units of rubble-mound breakwaters. The parameters of damage intensity function have been estimated through the time-dependent prediction of the expected cumulative damage level obtained from the sample path method. In particular, it is confirmed that the shock occurrences can be considered to be a discrete-time stochastic process by investigating the effects of uncertainty of the shock occurrences on the expected cumulative damage level with homogeneous Poisson process and doubly stochastic Poisson process that are the continuous-time stochastic processes. It can be also seen that the stochastic process of cumulative damage would depend directly on the design conditions, thus the preventive maintenance would be varied due to those. Finally, the optimal periods and scale for the preventive maintenance of armor units of rubble-mound breakwaters can be quantitatively determined with the failure limits, the levels of importance of structure, and the interest rates.

Interpretation of Interaction of Herbicides on Principal Paddy Weeds - By Use of Oxyfluorfen and Bensulfuron-methyl Data - (주요(主要) 논 잡초종(雜草種)에 대한 제초제간(除草劑間)의 상호작용효과(相互作用效果) 해석연구(解析硏究) - Oxyfluorfen과 Bensulfuron을 예(例)로 -)

  • Han, J.H.;Guh, J.O.;Chon, S.U.;Kwon, O.D.
    • Korean Journal of Weed Science
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    • v.12 no.2
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    • pp.144-157
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    • 1992
  • The study was conducted to compare the interprete methods and examine the feasibility of mixture use of oxyfluorfen and bensulfuron in controlling principal Paddy weeds, annuals and perennials. Application ratio of both chemicals were obtained from the combinations of 5 levels(0, 5, 10, 15, 20 g ai/ha) of each chemicals, respectively. All the treatments were applied at 5 days after transplanting and water was maintained at 3.0cm in depth. Shoot fresh-weight of weeds was assessed at 35 days after treatments. Data obtained was analysed by Colby, Isobole, Calculus, Regression and EQM method, respectively. The results from the analysis of variance on the principal weeds treated with oxyfluorfen and bensulfuron showed significant interactions at 1% level on both Echinochloa crus-galli and Eleocharis Kuroguwai, and total species at 0.5% level on both Potamogeton distinctus and Cyperus serotinus, but non significant on Scirpus juncoides and Sagittaria pygmaea. Thereafter, the results of the models applied to Echinochloa crus-galli, Eleocharis kuroguwai and total species were as follows ; 1. The Colby method gave values nearly identical to regression estimate method (both multiplicative models) as provided by Akobundu et al. The Colby method and Regression method indicated synergistic toward Echinochloa curs-galli, and total species, but antagonistic toward Eleocharis kuroguwai. 2. The Isobole method shows synergism on Echinochloa crus-galli at $ID_{50}$, and total species at $ID_{60}$ on Eleochari kuroguwai. 3. The Calculus method gave positive signs for the first differentiation and negative signs for the second differentiation except for some rates on Echinochloa crus-galli and total species, but reverse on Eleocharis kuroguwai. These result does not agree with the observed values. 4. ${\theta}$ value from the EQM method was greater than one at all combinations. This result was quite different from those of other methods. 5. The various models did not show the same results, but mixture of oxyfluorfen and bensulfuron tend to have synergistic effect. Weeding effect also was high. Treatment in terms of two chemical combination was expected to reduce rates, and to enhence weeding efficacy compared with single treatment.

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Analysis of Transfer Rate on Listeria monocytogenes Contaminated Pork Meat During Processing (돈육 가공공정 중 돈육에 오염된 Listeria monocytogenes의 전이율 분석)

  • Kim, Seong-Jo;Kim, Gwang-Hee;Park, Joong-Hyun;Park, Bo-Geum;Park, Myoung-Su;Oh, Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.27 no.4
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    • pp.432-441
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    • 2012
  • In this study, the transfer rate of wild type Listeria mon, ytogenes (LM) was investigated to establish the standard of safety management during pork meat pr, essing for meat to meat and meat to food contact surfaces contamination at 5 and $10^{\circ}C$. The transfer rate of LM from meat to meat during the pr, essing increased from 0.02% after 30 min to 0.42% after 120 min at $5^{\circ}C$, while for conveyor belt and stainless steel, it decreased from 0.015% and 0.013% after 30 min to 0.002% and 0.0003% after 120 min at $5^{\circ}C$, respectively (p < 0.05). When temperature increased to $10^{\circ}C$, the transfer rates of LM from meat to meat, conveyor belt and stainless steel were the highest at 60 min exposure, and all decreased after 120 min. In reverse, the transfer rate from food contact surface to pork meat was significantly higher than that from pork meat to food contact surface (p < 0.01). Also, the transfer rate to conveyor belt was significantly higher than stainless steel (p < 0.05) and it was highest at 30 min exposure time in both 5 and $10^{\circ}C$. This study indicates that the transfer and adherence rates of LM are influenced by the contact time and temperature. Consequently, these results were utilized to develop a predictive model with a high level of confidence which can lead to prevent cross-contamination during pork meat processing.

Isolation and Characterization of a Marine Bacterium, Pseudomonas sp. YJ-1 with Anti-Methicillin Resistant Staphylococcus aureus Activity (항 Methicillin Resistant Staphylococcus aureus 활성을 가지는 해양미생물 Pseudomonas sp. YJ-1의 분리와 특성)

  • Woo, Ye-Ju;Jeong, Seong-Yun
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.694-705
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    • 2017
  • The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

Comparative Analysis on Antioxidant Activity in Various Human Skin Fibroblasts and Mesenchymal Stem Cells (사람의 피부 섬유아세포 및 중간엽 줄기세포에서 항산화 활성의 반응에 대한 비교 분석)

  • Kong, Ji-Weon;Park, Ryeok;Park, Joon-Woo;Lee, Joo-Yeong;Choi, Yeon-Joo;Moon, Sun-Ha;Kim, Hyeon-Ji;Jeon, Byeong-Gyun
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.2
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    • pp.394-406
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    • 2019
  • The cellular senescence may be due to damage by the reactive oxygen species (ROS). This study has compared the antioxidant activity in the human cell lines of various origins, including 10S and 50S-derived normal skin fibroblasts, and 10S bone marrow, dental tissue and adipose-derived adult stem cells. After being exposed to $H_2O_2$, half inhibitory concentration ($IC_{50}$) values by cytotoxicity assay was significantly (P<0.05) lower in 50S-derived skin fibroblasts, than in 10S-derived skin fibroblasts and various adult stem cell lines. The cell population doubling time (PDT) and the cell frequency with high senescence associated-${\beta}$-galactose activity were remarkably increased in 50S-derived fibroblasts exposed to 50 ppm $H_2O_2$ for 7 days, than those of 10S-derived fibroblasts and various adult stem cell lines. Further, the expression level of antioxidant-related genes, glutathione peroxidase (GPX) and catalase (CAT), was investigated in 10S and 50S-derived skin fibroblasts, and 10S-derived various adult stem cells by reverse transcription polymerase chain reaction (RT-PCR). The expression level of GPX was higher in most of cell lines, compared to CAT, and a significantly (P<0.05) higher expression level of GPX was observed in 10S-derived skin fibroblasts and adult stem cell lines, compared to 50S-derived skin fibroblasts. We concluded that old-aged skin fibroblasts seemed to be less resistant against ROS than young-aged skin fibroblasts and adult stem cells.