• Title/Summary/Keyword: IS-PCR

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Characteristics of Coagulase-negative Staphylococci Isolates from Dental Clinic Environments in Busan, Korea (부산지역 치과환경에서 분리된 coagulase-negative staphylococci의 특성)

  • Jung, Hye-In;Jung, So Young;Park, Indal;Bae, Il Kwon
    • Journal of Life Science
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    • v.26 no.2
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    • pp.220-225
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    • 2016
  • Coagulase-negative staphylococci (CNS) have recently become the bacteria most frequently found in clinical infections. The aim of this study was to investigate the prevalence, antimicrobial susceptibilities, and molecular characteristics of CNS isolates from dental clinic environments in Busan, Korea. One hundred and fifty-four samples were collected from 10 dental clinics and dental hospitals in Busan from December 2014 to January 2015. Species were identified by matrix-assisted laser desorption/ionization–time-of-flight. Antimicrobial susceptibility was determined by disk diffusion methods. A polymerase chain reaction was performed to detect mecA, mupA gene, and SCCmec types. Of the 154 samples, 10(6.5%) isolates were identified as CNS (5 Staphylococcus epidermidis, 2 Staphylococcus capitis, 2 Staphylococcus, and 1 Staphylococcus haemolyticus). Among the 10 isolates, 6 were resistant to penicillin, 5 were resistant to gentamicin, 3 were resistant to tetracycline, and 2 were resistant to cefoxitin and erythromycin. However, clindamycin, ciprofloxacin, teicoplanin, and trimethoprim-sulfamethoxazole resistant isolates were not present. Genes encoding mecA were detected in 4 (2 S. warneri and 2 S. haemolyticus) isolates, and mupA in 1 (S. epidermidis) isolate. One methicillin-resistant CNS (S. warneri) isolate was determined as being of the SCCmec type I. It is concluded that CNS resistant to various antimicrobial agents was widely distributed in dental clinic environments in Korea.

Anti-proliferative and Pro-apoptotic Effects by Lees Extracts of Ehwa Makgeolli Containing Oriental Herbs (한방이화주 주박 추출물에 의한 암세포 항성장 및 세포사멸 기전 연구)

  • Kwon, Min-Jeong;Lee, Seung Hoon;Chung, Chung Wook;Sohn, Ho-Yong;Shin, Woo-Chang;Kim, Jong-Sik
    • Journal of Life Science
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    • v.26 no.2
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    • pp.241-246
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    • 2016
  • In the present study, ethanol extracts and their subsequent organic solvent fractions were extracted from the lees of Ehwa Makgeolli containing oriental herbs, a commercialized traditional Korean rice wine, and the prepared lees samples were designated as from KSD-E3-1 to KSD-E3-5. First, their effects on cell viability and on the expression of pro-apoptotic ATF3 and NAG-1 genes in human colorectal HCT116 cells were investigated. Among the treated lees samples, the hexane fraction (KSD-E3-2) and the ethyl acetate fraction (KSD-E3-3) of lees extracts from Ehwa Makgeolli significantly reduced cell viabilities, in a dose dependent manner. The treatment with KSD-E3-2 and KSD-E3-3 also increased the expression of pro-apoptotic NAG-1 and ATF-3 genes and their proteins, which were detected with RT-PCR and Western blot analysis, respectively. In addition, poly-(ADP-ribose) polymerase (PARP) cleavage was detected by treatment with the fraction KSD-E3-3, indicating that KSD-E3-3 could induce apoptosis in HCT116 cells. Interestingly, this PARP cleavage was recovered by transfection of NAG-1 small interfering RNA. The results indicate that NAG-1 is one of the genes responsible for apoptosis induced by the fraction KSD-E3-3 from Ehwa Makgeolli. Overall, the findings may help in understanding the molecular mechanisms of the anti-proliferative and pro-apoptotic activities mediated by the lees of Ehwa Makgeolli.

Effects of dietary polyphenol (-)-epigallocatechin-3-gallate on the differentiation of mouse C2C12 myoblasts (식이성 폴리페놀 (-)-epigallocatechin-3-gallate가 mouse C2C12 myoblast 분화에 미치는 영향)

  • Kim, Hye-Jin;Lee, Won-Jun
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.420-426
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    • 2007
  • In the present investigation, we studied the modulating effects of (-)-epigallocatechin-3-gallate(EGCG) on the differentiation of mouse C2C12 myoblasts. We found that the strong inhibitory effect of EGCG on DNA methyltransferase-mediated DNA methylation induced transdifferentiation of C2C12 myoblasts into smooth muscle cells demonstrated by both morphological changes and immunofluorescent staining. C2C12 myoblasts treated with EGCG for 4 days expressed smooth muscle ${\alpha}-actin$ protein. Real-time PCR data revealed that smooth muscle ${\alpha}-actin$ mRNA was induced by EGCG treated C2C12 myoblasts in a concentration-dependent manner. Smooth muscle ${\alpha}-actin$ mRNA concentration increased 330% and 490% after 2 and 3 days of 50 ${\mu}M$ of EGCG treatment. The expression of another smooth muscle marker, transgelin, mRNA was also increased up to 9-fold by 4 days of EGCG treatment compared with control in a concentration-dependent manner. These results suggested that C2C12 enables to transdifferentiate into smooth muscle when gene expression patterns are changed by the inhibition of DNA methylation induced by EGCG. In conclusion, transdifferentiation of C2C12 myoblasts into smooth muscle is resulted from the modulating effects of EGCG on DNA methylation which subsequently results in changing the expression pattern of several genes playing a critical role in the differentiation of C2C12 myoblasts.

The Effect of Haptoglobin on Expression of Inflammatory Cytokines in 3T3-L1 Preadipocytes. (3T3-L1 지방전구세포에서 합토글로빈에 의한 염증성 cytokine 발현 조절)

  • Cho, Jin-Kyung;Kim, Nam-Hoon;Oh, Mi-Kyung;Park, Seon-Joo;Kim, In-Sook
    • Journal of Life Science
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    • v.18 no.4
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    • pp.537-541
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    • 2008
  • White adipose tissue is now recognized as an important endocrine organ which secretes various signal factors and proteins termed 'adipokine'. Haptoglobin (Hp), which has been known as an acute phase protein, belongs to the adipokine. However, the function of Hp in adipose tissue remains unclear. To verify the role of Hp in preadipocytes, in this study, 3T3-L1 preadipocyte cells were stably transfected with human Hp gene and Hp-overexpressing cells were made. The Hp had no effect on cell growth of preadipocytes. By RT-PCR and Western blot analysis, the Hp inhibited gene expression of IL-6 and COX-2 and enhanced HO-1 synthesis in preadipocytes. Moreover, invasion assay showed the Hp suppressed migration of monocytes to preadipocytes. These findings suggest that the Hp may inhibit an inflammatory reaction in adipose tissue by regulating the expressions of pro-inflammatory and anti-inflammatory mediators, and by repressing monocytes/macrophages infiltration.

Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme (청국장으로부터 혈전용해 활성이 우수한 균주 분리 및 혈전용해효소정제)

  • Choi, Yeon Hee;Lee, Jun Seung;Bae, So Young;Yang, Keun Jae;Yeom, Kyu Won;Jo, Dong Hyeok;Kang, Ock Hwa;Baik, Hyung Suk
    • Journal of Life Science
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    • v.23 no.2
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    • pp.259-266
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    • 2013
  • To isolate the fibrinolytic enzyme, 268 strains from 21 samples were morphologically isolated from Cheonggukjang collected from Korea and Japan. Among the 268 strains, protease-producing bacteria were isolated in nutrient agar medium including 1% skimmed milk. As a result of this, 22 strains were isolated. Apiweb site was used to identify these strains based on their biochemical properties. In addition, 16S rRNA sequencing was performed to identify the strain. Most of the identified strains were Bacillus subtilis and B. amyloliquefaciens. Fibrinolytic enzyme activity was measured with the fibrin plate method. Five strains were finally selected: A2-14, A2-20, C1-05, C1-09, and F2-01. Of those five strains, the A2-20 strain, which is close to B. amyloliquefaciens, showed the strongest fibrinolytic activity. The fibrinolytic enzyme produced by the A2-20 strain was partially purified from culture supernatant by gel filtration and ion exchange chromatography. The optimal pH and temperature values of the partially purified enzyme were 7.0 and $35^{\circ}C$, respectively. Purified protein analysis was carried out with SDS-PAGE and zymography. A genetic analysis was also conducted by PCR based on the consensus sequence of fibrinolytic enzyme. Corresponding genes with a partial sequence of the A2-20 strain were identified.

Development of a simple and sensitive method to detect enteric viruses from oysters (굴로부터 장바이러스를 검출하기 위한 간단하고 민감한 방법의 개발)

  • Chung, Eun-Young;Je, Hee-Bok;Jun, Hong-Ki;Yoon, Jae-Deuk;Jee, Young-Mee;Cheon, Doo-Sung;Cho, Hae-Wol;Jang, Kyung-Lib
    • Journal of Life Science
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    • v.12 no.1
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    • pp.49-54
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    • 2002
  • Development of a rapid method possessing the requisite sensitivity and specificity for virus monitoring is necessary for protection of the shellfish-consuming public. Oysters tissue usually contains virus particles in relatively small concentrations along with various other substances that can interfere with detection steps. Therefore, the critical point concerning the detection of viruses in shellfish tissues resides in the processing of samples. The current study demonstrated the possibility of purifying small amounts of virus particles at the interface of a 10/50% sucrose gradient after a single round of sucrose gradient ultracentrifugation. We could detect HAV and poliovirus simultaneously from oyster tissues by using two different sets of primer. Furthermore, the method showed a high level of virus recovery rate (>95%) as determined by plaque assays of the final samples. Taken the advantages of the simple and sensitive methods, it was possible to detect 2 pfu of HAV in 5 g of oyster digestive tissues within 24h.

Delphinidin Suppresses Angiogenesis via the Inhibition of HIF-1α and STAT3 Expressions in PC3M Cells (전립선 암세포에서 delphinidin에 의한 HIF-1α와 STAT3 억제를 통한 혈관내피 성장 인자 발현 저해 효과)

  • Kim, Mun-Hyeon;Kim, Mi-Hyun;Park, Young-Ja;Chang, Young-Chae;Park, Yoon-Yub;Song, Hyun-Ouk
    • Korean Journal of Food Science and Technology
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    • v.48 no.1
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    • pp.66-71
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    • 2016
  • Delphinidin is a blue-red pigment and one of the major anthocyanins in plants. It plays an important role in anti-oxidant, anti-inflammatory, anti-mutagenic and anti-cancer properties. In this study, we investigated the inhibitory effects of delphinidin on vascular endothelial growth factor (VEGF) gene expression, an important factor involved in angiogenesis and tumor progression in human prostate cancer. Delphinidin decreased levels of epidermal growth factor (EGF)-induced VEGF mRNA expression in PC-3M cells. The expression of the EGF-induced hypoxia inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) and signaling transducer and activator of transcription 3 (STAT3) proteins, which are the major transcription factors for VEGF, were inhibited by delphinidin. In addition, delphinidin decreases HRE-promoter reporter gene activity, suggesting that delphinidin can suppress the transcription of HIF-$1{\alpha}$ under EGF induction, leading to a decrease in the expression of VEGF. Delphinidin specifically suppressed the phosphorylation of Akt, p70S6K, and 4EBP1, but not the phosphorylation of EGFR. Therefore, our results suggest that delphinidin may inhibit human prostate cancer progression and angiogenesis by inhibiting HIF-$1{\alpha}$, STAT3 and VEGF gene expression.

Inhibitory Effect of Rumex Crispus L. Fraction on Adipocyte Differentiation in 3T3-L1 Cells (소리쟁이 분획물의 지방세포 분화 억제 효과)

  • Park, Sung-Jin;Choi, Jun-Hyeok;Jung, Yeon-Seop;Yu, Mi Hee
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.90-96
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    • 2013
  • The anti-obesity effect of ethanol xtract and their fractions from Rumex Crispus L. on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated by suppressing adipocyte differentiation and lipid accumulation with Oil red O assay, western blot and real-time PCR analysis. Ethyl acetate fraction of Rumex crispus L. significantly inhibited adipocyte differentiation when treated during the adipocyte differentiation process, as assessed by measuring fat accumulation using Oil red O staining. In inducing differentiation of 3T3-L1 preadipocytes in the presence of an adipogenic cocktail, isobutylmethylxanthine (IBMX), dexamethasone- and insulin-along with ethyl acetate fraction residue processing treatment significantly decreased protein expression of obesity-related proteins, such as peroxisome-proliferators-activated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins ${\alpha}$ ($C/EBP{\alpha}$). These results indicate that ethyl acetate fraction of Rumex crispus L. is the most effective candidate for preventing obesity. However further studies will be needed to identify the active compounds that confer the anti-obesity activity of ethyl acetate fraction from Rumex crispus L.

Characterization of an isolated reovirus from the paradise fish Macropodus opercularis imported from Southeast Asia (동남아시아로부터 수입된 paradise fish Macropodus opercularis로부터 분리한 reovirus의 특성)

  • Kim, Wi-Sik;Kim, Soo-Jin;Kim, Jeong-Ho;Jung, Sung-Ju;Kim, Do-Hyung;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.335-342
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    • 2010
  • In 2008, mass mortality was observed in paradise fish Macropodus opercularis which was imported from Indonesia. PCR of these fish found positive for megalocytivirus and Mycobacterium sp., while an unidentified virus was culture-isolated using CHSE-214 cells. In the present study, we investigated characterization of the unidentified virus and its pathogenicity to determine whether the virus was the causative agent of the mass mortality of paradise fish. The unidentified virus induced cytopathic effect (CPE) with syncytia in CHSE-214 and other fish cells, BF-2, GF, SSN-1, FSP and FFN. The virus was resistant against treatments with IUdR, chloroform, acidity at pH 3, basicity at pH 11 and high temperature at $56^{\circ}C$ for 3h. By electron microscopy, the viral particles were spherical having a double capsid structure with approximately 65 nm in external diameter. Viral genome was composed of at least 10-segmented RNA with sizes ranging from 0.7 kb to 3.6 kb. Based on these characters, this virus can be classified into family Reoviridae. This reovirus did not cause any mortality in an artificial experiment conducted by injecting the virus to paradise fish. This indicates that the reovirus is not only responsible for the mass mortality of paradise fish in 2008.

Microbiological Quality Evaluation for Application of the HACCP System to the Bakery Products at Small Scale Bakeries (소규모 베이커리에서의 HACCP적용을 위한 미생물학적 위해도 평가)

  • 엄애선;권성희;정덕화;오상석;이헌옥
    • Korean journal of food and cookery science
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    • v.19 no.4
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    • pp.454-462
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    • 2003
  • Recently, the HACCP(Hazard Analysis Critical Control Point) system has been emphasized in food processing industries throughout the world. However, the system has, as yet, not been applied very well to domestic food industries. Due to the increase in the consumption of bakery products, more studies are required on the application of HACCP to establish the system in small-scale bakeries. This study was designed to provide basic data for setting management standards for HACCP, based on microbiological hazard evaluations of bakery products. Red bean paste filled breads, custard cream filled breads and cakes covered with fresh whipping cream were collected, and microbiological evaluations conducted on the raw materials, the manufacturing processes and potential hazards. The result showed the presence of coliforms in fresh cream of cakes and the soybean paste of soybean paste breads at levels as high as 105 CFU/g on the hazard analysis of the raw materials. Moreover, the general levels of bacteria and coliforms were over those of the standards during the intermediate fermentation and molding processes. Furthermore, high levels of coliforms were detected on the hands of the salesmen and bakers themselves. This suggests that the CCPs (Critical Control Points), such as fresh cream and red bean paste manufacturing process desperately require better management. There is also a requirement for education relating to personal hygiene for the production of hygienic bakery products and for the publics health.