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Analysis of Bacterial Diversity in Fermented Skate Using Culture-dependent and Culture-independent Approaches (배양 의존적 및 배양 비의존적 방법에 의한 홍어회 서식 미생물의 다양성 분석)

  • Lee, Eun-Jung;Kim, Tae-Hyung;Kim, Ha-Kun;Lee, Jung-Kee;Kwak, Hahn-Shik;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.322-328
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    • 2010
  • Fermented skate is a traditional Korean food popular in Southwestern area of Korea. It has a characteristic flavor and alkaline pH. In this study we tried to determine the microbial flora in fermented skate using two different approaches. In culture-independent method, we amplified V2 region of 16S rRNA gene by PCR and cloned them into pUC18 plasmid to construct 16S rDNA fragment library. BLAST searches for the sequences obtained from this library revealed that uncultured bacterium clone 054E11.b was the most dominant flora in this fermented fish. In culture-dependent method, we diluted suspension of skate and spreaded on MRS, PCA, and MacConkey plates. We identified colonies grown on those plates by using PCR amplification of V2 region of 16S rRNA and DNA sequencing. BLAST searches of those DNA sequences resulted in totally different species with the observations from the 16S rDNA library analysis. Discrepancies of results obtained from both approaches suggest that the agar plates used in culture-dependent method may be different from the real condition of fermented skate. Therefore, results from culture-independent approach using 16S rDNA fragment library analysis may reflect real microbial flora in fermented skate.

A Population Genetic Analysis of the Common Squid, Todarodes pacificus Steenstrup in the Korean Waters (한국 해역에 분포하는 오징어의 집단유전학적 방법에 의한 계군분석)

  • KANG Yong-Joo;KIM Yeong-Hye;HONG Yong-Ki;PARK Jung-Youn;PARK Kie-Young
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.3
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    • pp.320-331
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    • 1996
  • Electrophoretic comparisons of 482 individuals of Todarodes pacificus from 9 fishing areas were made to estimate genetic variability and differentiation using 17 enzymes. Strong activities were shown by 9 enzymes with 11 gene loci. The 9 sample lots could be divided into 3 genetic groups, based on dendrogram analysis using the Nei's genetic distance. The Summer, Autumn and Winter cohorts were identified as three seperated ecological populations which maintain genetic exchange. It is postulated that either the Summer cohort or the Autumn cohort has indepenently developed a local population that was isolated by hydrographic factors.

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Anti-skin Aging Potential of Alcoholic Extract of Phragmites communis Rhizome

  • Ha, Chang Woo;Kim, Sung Hyeok;Lee, Sung Ryul;Jang, Sohee;Namkoong, Seung;Hong, Sungsil;Lim, Hyosun;Kim, Youn Kyu;Sohn, Eun-Hwa
    • Korean Journal of Plant Resources
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    • v.33 no.6
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    • pp.604-614
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    • 2020
  • Chronological aging and photoaging affect appearance, causing wrinkles, pigmentation, texture changes, and loss of elasticity in the skin. Phragmites communis is a tall perennial herb used for its high nutritional value and for medicinal purposes, such as relief from fever and vomiting and facilitation of diuresis. In this study, we investigated the effects of ethanol extract of P. communis rhizome (PCE) on skin aging. The total flavonoid and total phenolic content in PCE were 2.92 ± 0.007 ㎍ of quercetin equivalents (QE) and 231.8 ± 0.001 ㎍ of gallic acid equivalents (GAE) per 100 mg of dried extract (n = 3). The half-maximal inhibitory concentration (IC50) values of PCE for 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and hydrogen peroxide scavenging activities were 0.96 and 0.97 mg/mL, respectively. PCE showed inhibitory effects on tyrosinase when L-tyrosine (IC50 = 1.25 mg/mL) and L-3,4-dihydroxyphenylalanine (IC50 = 0.92 mg/mL) were used as substrates. PCE treatment up to 200 ㎍/mL for 24 h did not cause any significant cytotoxicity in B16F10 melanocytes, human dermal fibroblasts (HDFs), and HaCaT keratinocytes. In B16F10 melanocytes, PCE (25 and 50 ㎍ /mL) inhibited melanin production and cellular tyrosinase activity after challenge with α-melanocyte-stimulating hormone (α-MSH; p < 0.05). In HDFs, PCE suppressed the mRNA expression of matrix metalloproteinase-1 (MMP-1) and reduced the activity of elastase (p < 0.05). In addition, ultraviolet B (UVB)-mediated downregulation of hyaluronic acid synthase-2 gene expression in HaCaT keratinocytes was also effectively suppressed by PCE treatment. Overall, our results showed that PCE has potential anti-skin aging activity associated with the suppression of hyperpigmentation, wrinkle formation, and reduction in dryness. PCE is a promising candidate for the development of an anti-skin aging cosmetic ingredient.

HIV-1 Tat-mediated protein transduction of human brain creatine kinase into PC12 cells

  • Jeong, Min-Seop;Kim, Dae-Won;Lee, Min-Jung;Lee, Yeom-Pyo;Kim, So-Young;Lee, Sun-Hwa;Jang, Sang-Ho;Lee, Kil-Soo;Park, Jin-Seu;Kang, Tae-Cheon;Cho, Sung-Woo;Kwon, Oh-Shin;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.41 no.7
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    • pp.537-541
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    • 2008
  • Epilepsy is characterized by the presence of spontaneous episodes of abnormal neuronal discharges and its pathogenic mechanisms remain poorly understood. Recently, we found that the expression of creatine kinase (CK) was markedly decreased in an epilepsy animal model using proteomic analysis. A human CK gene was fused with a HIV-1 Tat peptide to generate an in-frame Tat-CK fusion protein. The purified Tat-CK fusion protein was efficiently transduced into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced Tat-CK fusion protein was stable for 48 h. Moreover, the Tat-CK fusion protein markedly increased endogenous CK activity levels within the cells. These results suggest that Tat-CK provides a strategy for the therapeutic delivery of proteins in various human diseases including the delivery of CK for potential epilepsy treatment.

Differential Expression of Glucose Transporter Gene in Mouse Early Embryos (생쥐 초기배아의 Glucose Transporter유전자 발현 양상에 관한 연구)

  • Youm, Hye-Won;Byun, Hye-Kyung;Song, Gyun-Ji;Kim, Hae-Kwon;Lee, Ho-Joon
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.77-86
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    • 1998
  • The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture ($93.8{\pm}3.1$, n=35) is significantly higher than that of control and glucose-free group ($76.6{\pm}3.8$, n=35 and $68.2{\pm}4.3$, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.

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Modulation of Activator Protein-1 (AP-1) and MAPK Pathway by Flavonoids in Human Prostate Cancer PC3 Cells

  • Gopalakrishnan, Avanthika;Xu, Chang-Jiang;Nair, Sujit S.;Chen, Chi;Hebbar, Vidya;Kong, Ah-Ng Tony
    • Archives of Pharmacal Research
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    • v.29 no.8
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    • pp.633-644
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    • 2006
  • In last couple of decades the use of natural compounds like flavonoids as chemopreventive agents has gained much attention. Our current study focuses on identifying chemopreventive flavonoids and their mechanism of action on human prostate cancer cells. Human prostate cancer cells (PC3), stably transfected with activator protein 1 (AP-1) luciferase reporter gene were treated with four main classes of flavonoids namely flavonols, flavones, flavonones, and isoflavones. The maximum AP-1 luciferase induction of about 3 fold over control was observed with $20\;{\mu}M$ concentrations of quercetin, chrysin and genistein and $50\;{\mu}M$ concentration of kaempferol. At higher concentrations, most of the flavonoids demonstrated inhibition of AP-1 activity. The MTS assay for cell viability at 24 h showed that even at a very high concentration $(500\;{\mu}M)$, cell death was minimal for most of the flavonoids. To determine the role of MAPK pathway in the induction of AP-1 by flavonoids, Western blot of phospho MAPK proteins was performed. Four out of the eight flavonoids namely kaempferol, apigenin, genistein and naringenin were used for the Western Blot analysis. Induction of phospho-JNK and phospho-ERK activity was observed after two hour incubation of PC3-AP1 cells with flavonoids. However no induction of phospho-p38 activity was observed. Furthermore, pretreating the cells with specific inhibitors of JNK reduced the AP-1 luciferase activity that was induced by genistein while pretreatment with MEK inhibitor reduced the AP-1 luciferase activity induced by kaempferol. The pharmacological inhibitors did not affect the AP-1 luciferase activity induced by apigenin and naringenin. These results suggest the possible involvement of JNK pathway in genistein induced AP-1 activity while the ERK pathway seems to play an important role in kaempferol induced AP-1 activity.

Water Extract of Rosa laevigata Michx. Protects Hepatocytes from Arachidonic Acid and Iron-mediated Oxidative Stress (아라키돈산과 철 유도성 산화적 스트레스에 대한 금앵자(金櫻子) 열수 추출물의 간세포 보호 효능)

  • Ko, Hae Li;Jegal, Kyung Hwan;Song, Si Yeon;Kim, Nan Ee;Kang, Jiwon;Byun, Sung Hui;Kim, Young Woo;Cho, Il Je;Kim, Sang Chan
    • The Korea Journal of Herbology
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    • v.30 no.6
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    • pp.7-15
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    • 2015
  • Objectives : Rosa laevigata Michx. has been used for the treatment of renal disease in traditional Korean medicine. In this study, we investigated cytoprotective effect of R. laevigata water extract (RLE) against oxidative stress induced by arachidonic acid (AA) + iron.Methods : To evaluate the protective effects of RLE against AA + iron-induced oxidative stress in HepG2 cell, cell viability and changes on apoptosis-related proteins were assessed by MTT and immunoblot analyses. The effects of RLE on reduced glutathione level, production of reactive oxygen species and mitochondrial membrane potential were also monitored. Furthermore, to verify underlying molecular mechanism, NF-E2-related factor 2 (Nrf2) was examined by immunoblot analysis. Additionally, Nrf2 transactivation and its downstream target genes expression were also determined by reporter gene and realtime RT-PCR analyses.Results : RLE pretreatment (30-300 μg/ml) prevented cells from AA + iron-mediated cell death in a concentration dependent manner. In addition, 100 μg/ml RLE inhibited AA + iron-induced glutathione depletion, reactive oxygen species production and mitochondrial dysfunction. RLE accumulated nuclear Nrf2 and also transactivated Nrf2, which was evidenced by antioxidant response element- and glutathione S-transferase A2-driven luciferase activities and mRNA level of glutamate-cysteine ligase catalytic subunit, NAD(P)H:quinone oxidoreductase 1 and sestrin 2. Moreover, protective effect of RLE against AA + iron was abolished in Nrf2 knockout cells.Conclusions : These results indicate that RLE has the ability to protect hepatocyte against oxidative stress through Nrf2 activation.

A study on Resource Plant from Chollabuk-do Area for Developing Wild Flower (야생화 개발을 위한 전라북도 지역산 자원식물 조사)

  • 길봉섭;김영식;김창환;최성규;이종일
    • Korean Journal of Plant Resources
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    • v.12 no.1
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    • pp.43-49
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    • 1999
  • To develope wild flower collected from Chollabuk-do area resource plant species were investigated from May 1996 to April 1997. This study was focussed to make the basic data for better quality of breeding species from wild flower species and to keep proper gene pool for development with competitive power internationally in future. There was 131 species blooming in spring season(34.7%), 219 species in summer(58.1%), 25 species in autumn(6.6%) and 2 species in winter(0.5%), respectively. Flower color of all the plants was 26 kinds in diverse, among them, white flowers were occupied dominantly of 28.9%, yellow flowers, 20.7%, violet flowers, 16.4% and red purple flowers, 7.9%, respectively. Transplanted wild flower plants at the garden in the study area frequently were Chelidonium majus var. asiaticum, Hemerocallis fulva, Iris nertschinskia, Rhododendron yedoense var. poukhanense, Rhododendron mucronulatum, Pulsatilla koreana, Chrysanthemum zawadskii var. latilobum, Taraxacum platycarpum, Chrysanthemum boreale, Liriope platyphylla and Lycoris radiata. Twenty-six species of wild flowers such as Cymbidium goeringii, H. fulva, L. radiata, C. zawadskii var. latilobum, etc. were collected from their habitat and being selling at the market or road side. Total 343 taxa of wild flowers to prevent carrying out to foreign countries and to improve better quality were selected and recommended in this study, for example, they were 41 taxa of Compositae plant, 25 of Rosaceae, 20 of Liliaceae and 18 of Ranunculaceae in order. Acorus calamus var. angustatus, Cypripedium japonicum, C. macranthum, Calanthe discolor, Dicentra spectabilis, Ilex cornuta, Stewartia koreana, Abeliophyllum distichum, Forsythia saxatilis and Campanula glomerata var. dahurica should be protected as wild flowers because their numbers are decreasing remarkably, belong to rare species, being at a crisis to extinct species and important plants for research materials.

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Myxococcus xanthus socD500 mutation causes Sporulation and Induction of two C-signal Specific Genes (Myxococcus xanthus socD500에 의한 포자 형성 및 CsgA신호에 특이적 유전자의 발현에 관한 연구)

  • 이병욱
    • Journal of Life Science
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    • v.13 no.2
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    • pp.184-190
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    • 2003
  • Myxococcus xanthus is a Gram negative, rod-shaped, soil bacterium that displays a social behaviors, and multicellular development upon nutrient deprivation. The csgA gene encoding a cell surface protein is essential for developmental behaviors including rippling, aggregation, fruiting body formation and sporulation. csgA mutants show normal vegetative growth, but lack all these developmental phenotypes. Expression of the CsgA (C-signal) specific genes are eliminated or dramatically reduced in csgA mutants. In order to identify components of C-signal transduction pathway, second site mutations were introduced into csgA mutants and were identified which can fully or partially restore development of csgA mutants (Rhie, H. G. et. al. 1989. J. Bacteriol. 171, 3268-3276). One of such csgA suppressor mutations, socD500 restores only sporulation to csgA mutants at 15$^{\circ}C$. The socD500 mutaion however eliminates the three basic developmental requirements, starvation, high cell density and a solid surface. Only sporulation, not accompanied with fruiting body formation is induced simply by shifting the temperature of vegetatively growing cells from $32^{\circ}C$ to $15^{\circ}C$. Spores induced by socD500 mutation is not as thick as that of wild-type fruiting body. In socD500 genetic background, two of ten C-signal dependent genes, $\Omega$DK4506 and $\Omega$DK4406 are more highly expressed in growing cells at $15^{\circ}C$. These results indicate that the socD500 mutation may be partly involved in the regulation of expression of two C-signal dependent genes and genes for sporulation in this transduction pathway.

Probiotic Potential of Pediococcus pentosaceus BCNU 9070 (프로바이오틱 Pediococcus pentosaceus BCNU 9070 균주)

  • Shin, Hwa-Jin;Choi, Hye-Jung;Kim, Dong-Wan;Ahn, Cheol-Soo;Lee, Young-Geun;Jeong, Young-Kee;Joo, Woo-Hong
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1194-1200
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    • 2012
  • Lactic acid bacteria are generally recognized as beneficial probiotic organisms. Recent studies revealed that the potential of probiotic strains was essentially dependent on the bacterial-binding and adhesion capabilities to gut epithelial cells and the hydrophobicity of the cell surface. In this study, we screened some indigenous lactic acid bacteria from Kimchi and selected one lactic acid bacterium as a potential probiotic based on its cell surface hydrophobicity. Analysis of the 16S rRNA gene sequences of probiotic isolates indicated that the selected isolate (BCNU 9070 strain) was a member of Pediococcus pentosaceus. P. pentosaceus BCNU 9070 showed resistance to bile acids and acidic pH. The P. pentosaceus BCNU 9070 strain also inhibited the cell growth of six food-borne pathogens including Listeria monocytogenes and Shigella sonnei. In addition, the P. pentosaceus BCNU 9070 strain expressed bile salt hydrolase activity and showed an ability to assimilate cholesterol in vitro. On the basis of these results, P. pentosaceus BCNU 9070 is considered to have probiotic potential for applications in functional foodstuffs.