• Title/Summary/Keyword: Specific gene expression

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Identification of Microorganisms from Eggs in Hypermarket in the Northern Gyeonggi Area (경기 북부 일부 지역 대형 마트 유통계란에 오염된 미생물의 분리)

  • Chun, Myoung-Sook;Hong, Seung-Hee
    • The Korean Journal of Food And Nutrition
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    • v.22 no.3
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    • pp.396-401
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    • 2009
  • Microorganisms or their toxins can be transferred to eggs and cause food poisoning in humans. Therefore, this study was conducted to detect microbial contamination of eggs and to identify microorganisms in any contaminated eggs. Four different brands of eggs were collected from hypermarkets in the northern Gyeonggi area. The total bacterial counts on the shells of the eggs varied greatly between brands. In addition, various bacterial species including Klebsiella pneumoniae, Pseudomonas mendocina, Alcaligenes xylosoxidans, Alcaligenes faecalis, and Enterobacter cloacae were identified on eggshells. Furthermore, mean of total bacterial counts of four brands was $3.4{\times}10^4 cfu/m{\ell}$ and E. coli was detected on the eggshell of one brand egg. However, Salmonella was not identified on all brands of collected eggs. We also demonstrated that the E. coli isolated from the eggshell was not pathogenic based on the absence of pathogen-specific gene expression patterns. Taken together, the result of this study indicate that strict quality control and improved distribution controls are required to decrease microbial contamination and improve human health.

Research article Black ginseng activates Akt signaling, thereby enhancing myoblast differentiation and myotube growth

  • Lee, Soo-Yeon;Go, Ga-Yeon;Vuong, Tuan Anh;Kim, Jee Won;Lee, Sullim;Jo, Ayoung;An, Jun Min;Kim, Su-Nam;Seo, Dong-Wan;Kim, Jin-Seok;Kim, Yong Kee;Kang, Jong-Sun;Lee, Sang-Jin;Bae, Gyu-Un
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.116-121
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    • 2018
  • Background: Black ginseng (BG) has greatly enhanced pharmacological activities relative to white or red ginseng. However, the effect and molecular mechanism of BG on muscle growth has not yet been examined. In this study, we investigated whether BG could regulate myoblast differentiation and myotube hypertrophy. Methods: BG-treated C2C12 myoblasts were differentiated, followed by immunoblotting for myogenic regulators, immunostaining for a muscle marker, myosin heavy chain or immunoprecipitation analysis for myogenic transcription factors. Results: BG treatment of C2C12 cells resulted in the activation of Akt, thereby enhancing hetero-dimerization of MyoD and E proteins, which in turn promoted muscle-specific gene expression and myoblast differentiation. BG-treated myoblasts formed larger multinucleated myotubes with increased diameter and thickness, accompanied by enhanced Akt/mTOR/p70S6K activation. Furthermore, the BG treatment of human rhabdomyosarcoma cells restored myogenic differentiation. Conclusion: BG enhances myoblast differentiation and myotube hypertrophy by activating Akt/mTOR/p70S6k axis. Thus, our study demonstrates that BG has promising potential to treat or prevent muscle loss related to aging or other pathological conditions, such as diabetes.

Molecular Cloning of Plasmodium vivax Calcium-Dependent Protein Kinase 4

  • Choi, Kyung-Mi;Kim, Jung-Yeon;Moon, Sung-Ung;Lee, Hyeong-Woo;Sattabongkot, Jetsumon;Na, Byoung-Kuk;Kim, Dae-Won;Suh, Eun-Jung;Kim, Yeon-Joo;Cho, Shin-Hyeong;Lee, Ho-Sa;Rhie, Ho-Gun;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.48 no.4
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    • pp.319-324
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    • 2010
  • A family of calcium-dependent protein kinases (CDPKs) is a unique enzyme which plays crucial roles in intracellular calcium signaling in plants, algae, and protozoa. CDPKs of malaria parasites are known to be key regulators for stage-specific cellular responses to calcium, a widespread secondary messenger that controls the progression of the parasite. In our study, we identified a gene encoding Plasmodium vivax CDPK4 (PvCDPK4) and characterized its molecular property and cellular localization. PvCDPK4 was a typical CDPK which had well-conserved N-terminal kinase domain and C-terminal calmodulin-like structure with 4-EF hand motifs for calcium-binding. The recombinant protein of EF hand domain of PvCDPK4 was expressed in Echerichia coli and a 34 kDa product was obtained. Immunofluorescence assay by confocal laser microscopy revealed that the protein was expressed at the mature schizont of P. vivax. The expression of PvCDPK4-EF in schizont suggests that it may participate in the proliferation or egress process in the life cycle of this parasite.

Raw Starch-digesting Amylase is Comprised of two Distinct Domains of Catalytic and Substrate-Adsorbable Domain: Role of the C- Terminal Region in Raw-Starch-Binding

  • Kim, Cheorl-Ho
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.40-45
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    • 2001
  • Raw starch-digesting amylase (BF-2A, M.W. 93, 000 Da) from Bacillus circulans F-2 was converted to two components during digestion with subtilisin. Two components were separated and designated as BF-2A' (63, 000 Da) and BF-2B (30, 000 Da), respectively. BF-2A' exhibited the same hydrolysis curve for soluble starch as the original amylase (BF-2A). Moreover, the catalytic activities of original and modified enzymes were indistinguishable in $K_{m}$, Vmax for, and in their specific activity for soluble starch hydrolysis. However, its adsorbability and digestibility on raw starch was greatly decreased. Furthermore, the enzymatic action pattern on soluble starch was greatly different from that of the BF-2A. A smaller peptide (BF-2B) showed adsorb ability onto raw starch. By these results, it is suggested that the larger peptide (BF-2A') has a region responsible for the expression of the enzyme activity to hydrolyze soluble substrate, and the smaller peptide (BF-2B) plays a role on raw starch adsorption. A similar phenomenon is observed during limited proteinase K, thermolysin, and endopeptidase Glu-C proteolysis of the enzyme. Fragments resulting from proteolysis were characterized by immunoblotting with anti-RSDA. The proteolytic patterns resulting from proteinase K and subtilisin were the same, producing 63- and 30-kDa fragments. Similar patterns were obtained with endopeptidase Glu-C or thermolysin. All proteolytic digests contained a common, major 63-kDa fragment. Inactivation of RSDA activity results from splitting off the C-terminal domain. Hence, it seems probable that the protease sensitive locus is in a hinge region susceptible to cleavage. Extracellular enzymes immunoreactive toward anti-RSDA were detected through whole bacterial cultivation. Proteins of sizes 93-, 75-, 63-, 55-, 38-, and 31-kDa were immunologically identical to RSDA. Of these, the 75-kDa and 63-kDa proteins correspond to the major products of proteolysis with Glu-C and thermolysin. These results postulated that enzyme heterogeneity of the raw starch-hydrolysis system might arise from the endogeneous proteolytic activity of the bacterium. Truncated forms of rsda, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional amylase that did not bind to raw starch. This indicates that the conserved region of RSDA constitutes a raw starch-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.d.

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Increment of fructan biosynthesis in rice by transformation of 1-sst and 1-fft genes isolated from jerusalem artichoke (Helianthus tuberosus L.) (돼지감자 유래 1-sst와 1-fft 유전자의 형질전환 발현에 의한 벼의 fructan 생합성 증진)

  • Kang, Kwon-Kyoo;Song, Beom-Heon;Lee, Gyong-A;Lee, Hye-Jung;Park, Jin-Ha;Jung, Yu-Jin;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.102-109
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    • 2010
  • Fructan has been found to accumulate in various tissues during periods when light levels increased carbon fixation where low temperatures reduced growth rates while photosynthesis continued. In this study, we have cloned 1-sucrose:sucrose fructosyl transferase(1-sst) and 1-fructan: fructan fructosyl transferase (1-fft, a key enzyme for the synthesis of fuctan) from Jerusalem Artichoke (Helianthus tuberosus L.). The recombinant vector with 1-sst and 1-fft has been constructed under the control of 35S promoter of KJGV-B2 vector and transgenic plants obtained by Agrobacterium tumefaciens LBA4404. PCR analysis carried out on the putative transgenic plants for amplification of the coding region of specific gene (1-sst, 1-fft), and HPT genes. Transgenic lines carrying of 1-sst and 1-fft were confirmed for integration into the rice genome using Southern blot hybridization and RT-PCR. The transgenic plants in $T_2$ generation were selected and expression pattern analysis revealed that 1-sst and 1-fft were stable. This analysis confirmed the presence of low-molecular-weight fructan in the seedling of the transgenic rices. Therefore, cold tolerance and carbohydrate metabolism will be possible to develop resistant plants using the transgenic rice.

Efficacy of callus induced from Ullengdo niche plants for skin protection (식물세포배양기술을 이용한 울릉도 자생식물의 세포주 개발 및 피부세포 효능)

  • Choi, Yun Hui;Jung, Hae Soo;Cho, Moon Jin;Song, Mi Young;Seo, Hyo Hyun;Moh, Sang Hyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.8
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    • pp.5070-5077
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    • 2014
  • Many countries in the world have protected their native plants and utilized them as industrial materials in each country. In this aspect, it is increasingly important to develop cosmetics materials using native plants in Korea. Cosmetic materials have been developed with niche plants, such as Campanula takesimana Nakai, Dianthus superbus, Aster spathulifolius in Ullengdo, in which a specific plant distribution by distinct climate and environment was present. Water and ethanol extractions were performed from the calluses of Campanula takesimana Nakai, Dianthus superbus, Aster spathulifolius. HPLC analysis revealed different compositions and functions of effective elements in each ethanol extract. For example, all types of ethanol extracts showed an ability to heal wounds. In particular, the expression of the inflammation-related gene, COX-2, was decreased in response to the ethanol extracts of Dianthus superbus. These results indicate that the ethanol extracts from niche plants' calluses in Ullengdo are natural and environmentally-friendly compounds, and can be used as medical supplies associated with anti-inflammation and wound healing.

Enzymatic Characterization of Bacillus cereus Lactate Dehydrogenase Isozymes Expressed in Escherichia coli (Bacillus cereus에서 유래한 Lactate Dehydrogenase 동질효소 유전자의 대장균 내 발현 및 효소특성 규명)

  • Jang, Myoung-Uoon;Park, Jung-Mi;Lee, Hong-Gyun;Lee, So-Ra;Kim, Tae-Jip
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.213-218
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    • 2010
  • Lactate dehydrogenases (LDHs) have been highly focused for long time, due to their important roles in biochemical and metabolic pathways of cells. On the basis of genome-wide searching results, three putative LDH genes from Bacillus cereus ATCC 14579 genome have been PCR-amplified, cloned, and well-expressed in E. coli. All three BcLDH isozymes are supposed to share highly conserved catalytic amino acid residues in common $NAD^+$-dependent LDHs. Meanwhile, BcLDH1 consisting of 314 amino acids shares 86 and 49% of identities with BcLDH2 and 3, respectively. Interestingly, only BcLDH1 showed the converting activities between L-lactate and pyruvate in the presence of $NAD^+$ coenzyme, while the other isozymes are likely to have almost no activity. As a result, it was revealed that BcLDH1 can be a typical $NAD^+$-dependent L-lactate-specific dehydrogenase.

Ethanol Extracts of Citrus Peel Inhibits Adipogenesis through AMPK Signaling Pathway in 3T3-L1 Preadipocytes (진피 에탄올 추출물의 AMPK signaling pathway를 통한 3T3-L1 지방전구세포의 adipogenesis 억제에 관한 연구)

  • Jo, Hyun Kyun;Han, Min Ho;Hong, Su Hyun;Choi, Yung Hyun;Park, Cheol
    • Journal of Life Science
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    • v.25 no.3
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    • pp.285-292
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    • 2015
  • Citrus peel (CP) is used as a traditional herb with diverse beneficial pharmacological activities, such as anti-inflammatory, anti-oxidant, and anti-allergic effects. However, the anti-obesity effects of citrus peel are poorly defined. The aim of this study was to evaluate ethanol extracts of citrus peel (EECP) for its adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes. The aim of this study was to evaluate an EECP for its adipocyte differentiation and adipogenesis in 3T3-L1 preadipocytes. Treatment with EECP significantly suppressed the terminal differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by a decrease in lipid droplet number and lipid content and an accumulation of cellular triglyceride. EECP exhibited potential adipogenesis inhibition and downregulated the expression of pro-adipogenic transcription factors, such as sterol regulatory elementbinding protein-1c (SREBP-1c), peroxisome proliferator-activated receptor-γ (PPARγ), CCAAT/enhancerbinding proteins α (C/EBPα) and C/EBPβ, and adipocyte expressed genes, such as adipocyte fatty acid binding protein (aP2) and Leptin. In addition, EECP treatment effectively activated the AMP-activated protein kinase (AMPK) signaling pathway; however, compound C, a specific inhibitor of AMPK, significantly reduced the EECP-induced inhibition of adipogenesis. Taken together, these results indicate EECP showed strong anti-obesity effects through the AMPK signaling pathway, and further studies will be needed to identify the active compounds that confer the anti-obesity activity of EECP.

5-Aza-2'-deoxycytidine Inhibits the Maintenance of Cancer Stem Cell in a Mouse Model of Breast Cancer (마우스 유방암 모델에서 5-Aza-2'-deoxycytidine의 암줄기세포 유지 억제 효과)

  • Nho, Kyoung-Jin;Yang, In-Sook;Kim, Ran-Ju;Kim, Soo-Rim;Park, Jeong-Ran;Jung, Ji-Youn;Cho, Sung-Dae;Nam, Jeong-Seok
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1164-1169
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    • 2009
  • Aberrant DNA methylation plays an important role in the development of cancer. It has been reported recently that DNA hypermethylation is involved in the maintenance of cancer stem cells. The present study was designed to test the hypothesis that the demethylating agent, 5-aza-2'-deoxycytidine (AZA), can inhibit the potential for maintenance of cancer stem cells. To validate this hypothesis, we used 4T1 syngeneic mouse models of breast cancer. The AZA pre-treated 4T1 cells showed a dramatic inhibition of tumorsphere formation, compared to their counterparts in vitro. In addition, the AZA treatment significantly suppressed the expression of stem regulator genes, such as oct-4, nanog and sox2, compared to counterparts in vivo. Therefore, selective inhibition of DNA methylation may be useful for stem-specific cancer therapy.

Upregulation of miR-23b Enhances the Autologous Therapeutic Potential for Degenerative Arthritis by Targeting PRKACB in Synovial Fluid-Derived Mesenchymal Stem Cells from Patients

  • Ham, Onju;Lee, Chang Youn;Song, Byeong-Wook;Lee, Se-Yeon;Kim, Ran;Park, Jun-Hee;Lee, Jiyun;Seo, Hyang-Hee;Lee, Chae Yoon;Chung, Yong-An;Maeng, Lee-So;Lee, Min Young;Kim, Jongmin;Hwang, Jihwan;Woo, Dong Kyun;Chang, Woochul
    • Molecules and Cells
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    • v.37 no.6
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    • pp.449-456
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    • 2014
  • The use of synovial fluid-derived mesenchymal stem cells (SFMSCs) obtained from patients with degenerative arthropathy may serve as an alternative therapeutic strategy in osteoarthritis (OA) and rheumatoid arthritis (RA). For treatment of OA and RA patients, autologous transplantation of differentiated MSCs has several beneficial effects for cartilage regeneration including immunomodulatory activity. In this study, we induced chondrogenic differentiation of SFMSCs by inhibiting protein kinase A (PKA) with a small molecule and microRNA (miRNA). Chondrogenic differentiation was confirmed by PCR and immunocytochemistry using probes specific for aggrecan, the major cartilaginous proteoglycan gene. Absorbance of alcian blue stain to detect chondrogenic differentiation was increased in H-89 and/or miRNA-23b-transfected cells. Furthermore, expression of matrix metalloproteinase (MMP)-9 and MMP-2 was decreased in treated1 cells. Therefore, differentiation of SFMSCs into chondrocytes through inhibition of PKA signaling may be a therapeutic option for OA or RA patients.