• Title/Summary/Keyword: Non-Isolation

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Overexpression of the MUC1 Gene in Iranian Women with Breast Cancer Micrometastasis

  • Mansouri, Neda;Movafagh, Abolfazl;Soleimani, Shahrzad;Taheri, Mohammad;Hashemi, Mehrdad;Pour, Atefeh Heidary;Shargh, Shohreh Alizadeh;Mosavi-Jarahi, Alireza;Sasaninejad, Zahra;Zham, Hanieh;Hajian, Parastoo;Moradi, Hossein Allah;Mirzaei, Hamid Reza;Fardmanesh, Hedieh;Ohadi, Mina
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.sup3
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    • pp.275-278
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    • 2016
  • The membrane epithelial mucin MUC1 is expressed at the luminal surface of most simple epithelial cells, but expression is greatly increased in most breast cancers. The aims of present study were to investigate expression of the MUC1 gene and interactive affects in metastases. Whole cell RNA isolation from 50 sentinel lymph nodes (SNLs) of breast cancer patients was performed using reverse transcription and real-time PCR. All patients were diagnosed with breast cancer and without metastasis, confirmed by IHC staining. The evaluation of tumor and normal samples for expression of MUC1 gene, the results were 49.1% non-expressive and 45.3% expression (Student t, p = 0.03). Also in comparison of normal breast tissue and breast cancer SLN for MUC1 gene, MUC1 negative SLNs were 75.0% (18 samples) and MUC1 positive samples were 25.0% (6 samples). Over-expression of MUC1 gene may offer a target for therapy related to progression and metastasis in women with breast cancer.

Human Being in the Contemporary Society (도시적 인간상 연구 - 본인 작품을 중심으로-)

  • 박성원
    • Proceedings of the Korea Contents Association Conference
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    • 2004.05a
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    • pp.553-561
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    • 2004
  • It works as intermediation of communication of publics these days. Since 20 century, We, Koreans, have established new chaotic multi culture with traditional Korean culture and other different culture from everywhere. Meanwhile, we occupied the most powerful semi-conduct and IT indusry. Within those circumstance, people feel very confused in political, cultural and social aspect. The society armed with economy and popularization promotes material satisfaction with this potential possibility of anonymous masses. However, it results to cause loneliness, isolation, alienation, anonymity, non individuality and commodity of culture. In my work, such phenomenon reveals through human character in a city. People are exposed culture of consumption and surrounded and tempted by all those artificial and superficial atmosphere. Human are possessed and exposed to attractive products and visual images. Finally they make themselves stuck in their case of this world. People lose their own identify and shape of bodies. That is our portrait, who are living this moment. Also, this is a symbol that destroys this modern society. As a result, 1 consider such aspects through those elements above to think how to keep and rethink our identity and what to do for this world.

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Lipase Inhibitory Mode of Dieckol Isolated from Eisenia bicyclis Ethanol Extract (Eisenia bicyclis 에탄올 추출물로부터 분리한 Dieckol의 Lipase 저해 Mode)

  • Jung, Seul-A;Kim, Koth-Bong-Woo-Ri;Kim, Dong-Hyun;Cho, Ji-Young;Kim, Tae-Wan;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.112-118
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    • 2013
  • This study was performed to investigate the possible use of Eisenia bicyclis (EB) ethanol extract to inhibit activity against lipase. In tests, the lipase inhibitory activity of EB ethanol extract was noted as being 43, 27, and 24% at concentrations of 5, 2.5, and 1 mg/ml, respectively. Isolation was carried out by liquid and liquid extraction, silica-gel column chromatography, and HPLC. The results showed that the lipase inhibitory activity of the ethyl acetate (EA) fraction from EB ethanol extract exhibited the strongest lipase inhibitory activity with an $IC_{50}$ value of 1.31 mg/ml. The EA fraction was separated using silica-gel column chromatography and we obtained 22 sub-fractions. Amongst them, the EA1 fraction showed the highest lipase inhibitory activity with an $IC_{50}$ value of 0.54 mg/ml. Eight peaks were obtained from the EA1 fraction by HPLC. Fraction 5 also showed a strong lipase inhibitory activity with an $IC_{50}$ value of 0.37 mg/ml. The fraction 5 was identified as dieckol and the inhibition pattern analyzed from Lineweaver-Burk plots revealed a non-competitive inhibitor. These results suggest that EB has potential as a natural anti-obesity agent.

Isolation of Probiotic Piliated Lactobacillus rhamnosus Strains from Human Fecal Microbiota Using SpaA Antiserum-Based Colony Immunoblotting

  • Yang, Zhen-quan;Xue, Yu;Rao, Sheng-qi;Zhang, Mi;Gao, Lu;Yin, Yong-qi;Chen, Da-wei;Zhou, Xiao-hui;Jiao, Xin-an
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.1971-1982
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    • 2017
  • Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect $2.5{\times}10^3CFU/ml$ of pLR colonies spiked in $10^6CFU/ml$ of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.

Isolation of CONSTANS as a TGA4/OBF4 Interacting Protein

  • Song, Young Hun;Song, Na Young;Shin, Su Young;Kim, Hye Jin;Yun, Dae-Jin;Lim, Chae Oh;Lee, Sang Yeol;Kang, Kyu Young;Hong, Jong Chan
    • Molecules and Cells
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    • v.25 no.4
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    • pp.559-565
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    • 2008
  • Members of the TGA family of basic domain/leucine zipper transcription factors regulate defense genes through physical interaction with NON-EXPRESSOR OF PR1 (NPR1). Of the seven TGA family members, TGA4/octopine synthase (ocs)-element-binding factor 4 (OBF4) is the least understood. Here we present evidence for a novel function of OBF4 as a regulator of flowering. We identified CONSTANS (CO), a positive regulator of floral induction, as an OBF4-interacting protein, in a yeast two-hybrid library screen. OBF4 interacts with the B-box region of CO. The abundance of OBF4 mRNA cycles with a 24 h rhythm under both long-day (LD) and short-day (SD) conditions, with significantly higher levels during the night than during the day. Electrophoretic mobility shift assays revealed that OBF4 binds to the promoter of the FLOWERING LOCUS T (FT) gene, a direct target of CO. We also found that, like CO and FT, an OBF4:GUS construct was prominently expressed in the vascular tissues of leaf, indicating that OBF4 can regulate FT expression through the formation of a protein complex with CO. Taken together, our results suggest that OBF4 may act as a link between defense responses and flowering.

Multiple Confirmation and RAPD-genotyping of Enterobacter sakazakii Isolated from Sunsik (선식에서 분리한 Enterobacter sakazakii의 복합동정 및 RAPD를 이용한 genotyping)

  • Choi, Jae-Won;Kim, Yun-Ji;Lee, Jong-Kyung;Kim, Young-Ho;Kwon, Ki-Sung;Hwang, In-Gyun;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.40 no.1
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    • pp.101-105
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    • 2008
  • Enterobacter sakazakii is implicated in severe forms of neonatal infections such as meningitis and sepsis. This organism has been isolated from a wide range of foods, including cheese, vegetables, grains, herbs, and spices, but its primary environment is still unknown. Generally, dried infant milk formula has been epidemiologically identified as the source of E. sakazakii. Sunsik (a powdered mixture of roasted grains and other foodstuffs) is widely consumed in Korea as a side dish or energy supplement. Sunsik is consumed without heat treatment; thus, lacking an additional opportunity to inactivate foodborne pathogens. Therefore, its microbiological safety should be guaranteed. In this study, the prevalence of E. sakazakii was monitored in 23 different sunsik component flours, using FDA recommended methods; but E. sakazakii medium (Neogen) and Chromogenic E. sakazakii medium (Oxoid) were used as the selective media. In total, presumptive E. sakazakii strains were isolated from 8 different sunsik powders. Subsequently, an API 20E test was conducted, and 15 strains from 5 different sunsik flours (sea tangle, brown rice, non-glutinous rice, cheonggukjang, dried anchovy) were confirmed as E. sakazakii. Fifteen strains were again confirmed by PCR amplification, using three different primer sets (tDNA sequence, ITS sequence, 16S rRNA sequence), and compared to ATCC strains (12868, 29004, 29544, 51329). They were once again confirmed by their enzyme production profiles using an API ZYM kit. Finally, RAPD (random amplified polymorphic DNA)-genotyping was carried out as a monitoring tool to determine the contamination route of E. sakazakii during processing.

Phospholipids Isolation from Squid Viscera Residues After Supercritical Carbon Dioxide Extraction (오징어 내장의 초임계 이산화탄소 추출 잔류물로부터 인지질의 분리)

  • U, Pyoung-Ook;Chun, Byung-Soo
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.741-746
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    • 2010
  • Phospholipids were recovered from squid viscera residues by ethanol extraction after supercritical carbon dioxide($SCO_2$) extraction and from squid viscera was not processed $SCO_2$ by various organic solvent extraction. $SCO_2$ extraction were performed at $45^{\circ}C$ and 20 MPa for removal of non polar lipid molecules from freeze dried squid viscera sample. Phospholipids were extracted from freeze dried squid viscera sample by chloroform, hexane, methanol, and ethanol and from $SCO_2$extracted squid viscera sample by ethanol. The pH was fixed at 5.7 for all phospholipids extraction conditions. Phospholipid classes were analyzed by HPLC equipped with evaporative light scattering detector (ELSD). Phosphatidyl choline(PC) extracted by ethanol from $SCO_2$ extracted residues was higher than that of extracted by ethanol from squid viscera. But phosphatidyl ethanolamine(PE) and phosphatidic acid(PA) were extracted higher percentage in raw squid viscera. The fatty acid compositions in phospholipids extract by ethanol extract from $SCO_2$ extracted residues were analyzed by gas chromatography(GC). Docosahexanoic acid(DHA) was found in highest percentage in phospholipid extract.

Isolation and identification of soycurd forming lactic acid bacteria which produce GABA from kimchi (김치로부터 GABA를 생산하는 커드 형성 젖산균의 분리 및 동정)

  • Kim, Eun-Ah;Mann, So-Yon;Kim, Su-In;Lee, Ga-Young;Hwang, Dae-Youn;Son, Hong-Joo;Lee, Chung-Yeol;Kim, Dong-Seob
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.705-711
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    • 2013
  • Gamma amino butyric acid (GABA), known as a non-protein amino acid and major inhibitory neurotransmitter in the brain, has several functional properties such as neurotransmission, induction of hypotension, tranquilizer, and diuretic effects. The purpose of this study was to isolate and identify lactic acid bacteria, producing high GABA in fermented soy curd. Thirty-two strains of tofu-forming lactic acid bacteria were isolated from kimchi which a traditional Korean food fermented with many kind of microorganism. Among 32 strains, four strains (strain No. 10, 104, 214, 249) formed firm soycurd. In order to select lactic acid bacteria having high GABA producing potential, the isolated strains were cultured in the soymilk and fermented for 48 hr at $37^{\circ}C$. A strain No. 383, which showed highest GABA contents in fermented soycurd, was identified as L. sakei by 16S rDNA sequencing and API analysis, and named as L. sakei 383. L. sakei 383 showed optimal growth up to 24 hr at $35^{\circ}C$ in MRS broth. The optimal time and temperature for GABA production were 18 hr and $35^{\circ}C$ in soymilk. In the optimal condition time and temperature, GABA content of fermented soycurd by L. sakei 383 was 8.65 mg/100 g.

Isolation of Microorganisms and Development of Microbial Augmentation for Treatment of Paper Mill Wastewater (제지폐수 처리용 미생물의 분리 및 복합 미생물제제의 개발)

  • Kang, Dae-Ook;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.21 no.4
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    • pp.554-560
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    • 2011
  • This study was performed to investigate the effects of microbial augmentation on the biological treatment of paper mill wastewater. Three bacteria (KN11, KN13, KN27) capable of degrading aromatic compounds and a bacterial strain (GT21) producing an extracellular cellulase were isolated from soil and wastewater by selective enrichment culture. Through morphological, physiological, and biochemical taxonomies, isolated strains of KN11, KN13, KN27, and GT21 were identified as Acinetobacter sp., Neisseria sp., Bacillus sp., and Pseudomonas sp. and named Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, respectively. For analysis of non-biodegradable and chemical oxygen demand (COD)-increasing matter in a paper mill wastewater, we utilized GC/MS to detect aromatic compounds and their derivatives containing several substituted functional groups. The microbial augmentation, J30 formulated with the mixture of bacteria including Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, was used for the treatment of paper mill wastewater. The optimum temperature and pH for COD removal of the microbial augmentation, J30, were $30^{\circ}C$ and 7.5, respectively. For evaluation of the industrial applicability of the microbial augmentation, J30 in the pilot test, treatment efficiency was examined using paper mill wastewater. The microbial augmentation, J30, showed a COD removal rate of 87%. On the basis of the above results, we designed the wastewater treatment process of the activated sludge system.

Isolation and Characterization of a Diesel-Degrading Bacterium, Gordonia sp. SD8 (디젤 분해 세균 Gordonia sp. SD8 분리 및 특성)

  • Hong, Sun-Hwa;Kim, Ji-Young;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.335-339
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    • 2010
  • A diesel-degrading bacterium, Gordonia sp. SD8, was isolated from soil contaminated with petroleum, and its diesel degradation was characterized in a soil as well as a liquid culture system. SD8 could grow in the mineral salt medium supplemented with diesel as a sole carbon and energy source. The maximum specific growth rate ($0.67{\pm}0.05\;d^{-1}$) and diesel degradation rate ($1,727{\pm}145$ mg-TPH $L^{-1}\;d^{-1}$) of SD8 showed at 20,000 mg-TPH $L^{-1}$ and $30^{\circ}C$, and then this bacterium could degrade high strength of diesel of 40,000 mg-TPH $L^{-1}$. The residual diesel concentration in the inoculated soil with SD8 was 3,724 mg-TPH kg-dry $soil^{-1}$ after 17 days, whereas the diesel concentration in the non-inoculated soil was $8,150{\pm}755$ mg-TPH kg-dry $soil^{-1}$. These results indicate that Gordonia sp. SD8 can serve as a promising microbial resource for the bioremediaion of contaminated soil with petroleum hydrocarbons including diesel.