• The Korean Journal of Physiology and Pharmacology
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• 제19권6호
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• pp.549-555
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• 2015

We attempted to investigate molecular mechanisms underlying phenotypic change of vascular smooth muscle cells (VSMCs) by determining signaling molecules involved in chemokine production. Treatment of human aortic smooth muscle cells (HAoSMCs) with thrombin resulted not only in elevated transcription of the (C-C motif) ligand 11 (CCL11) gene but also in enhanced secretion of CCL11 protein. Co-treatment of HAoSMCs with GF109230X, an inhibitor of protein kinase C, or GW5074, an inhibitor of Raf-1 kinase, caused inhibition of ERK1/2 phosphorylation and significantly attenuated expression of CCL11 at transcriptional and protein levels induced by thrombin. Both Akt phosphorylation and CCL11 expression induced by thrombin were attenuated in the presence of pertussis toxin (PTX), an inhibitor of Gi protein-coupled receptor, or LY294002, a PI3K inhibitor. In addition, thrombin-induced production of CCL11 was significantly attenuated by pharmacological inhibition of Akt or MEK which phosphorylates ERK1/2. These results indicate that thrombin is likely to promote expression of CCL11 via PKC/Raf-1/ERK1/2 and PTX-sensitive protease-activated receptors /PI3K/Akt pathways in HAoSMCs. We propose that multiple signaling pathways are involved in change of VSMCs to a secretory phenotype.

• 한국임상약학회지
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• 제28권3호
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• pp.174-180
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• 2018

Objective: Clostridium difficile Infection (CDI) is one of the common nosocomial infections. As elderly population increases, the proper treatment has been emphasized. We investigated the risk factors associated with CDI unimprovement in elderly patients. Furthermore, we performed drug use evaluation of old CDI patients and oldest-old CDI patients. Methods: It was a retrospective study using electronic medical record at Kangbuk Samsung Medical Center (KBSMC) from January 2016 to December 2017. Seventy three patients aged 65 years or older, diagnosed with CDI by Clostridium difficile Toxin B Gene [Xpert] were screened and they were assessed for risk factors regarding unimprovement status. We also evaluated drug use evaluation in old patients ($65{\leq}age$<80) and oldest-old patients ($80{\leq}age$) by assessing the use of initial therapy, severity, dose, route, treatment course, days of use, total days of use and treatment outcome of initial therapy. Results: Out of 73 patients aged over 65 years, four patients were excluded because they did not receive any treatment. There were 31 improved patients and 38 unimproved patients after initial therapy. We were able to find out patients with surgical comorbidity or endocrine comorbidity (especially, diabetes mellitus) had 2.885 more risk of becoming unimproved than those patients without surgical comorbidity or endocrine comorbidity. Drug use evaluation for CDI was generally fair, but vancomycin as initial therapy is more recommended than metronidazole. Conclusion: Although age, antibiotics exposure, use of antacids are all important risk factors for CDI, our result did not show statistical significance for these risk factors. However, the study is meaningful because the number of elderly population keeps increasing and recently updated guideline suggests the use of vancomycin as drug of choice for CDI.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.5063-5067
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• 2012

New molecular markers of cancer had emerged with novel applications in cancer prevention and therapeutics, including for breast cancer of unknown causes, which has a high impact on the health of women worldwide. The purpose of this research was to detemine protein and mRNA expression of synaptic vesicle 2 (SV2) isoforms A, B and C in breast cancer cell lines. Cultured cell lines MDA-MB-231, SKBR3, T47D were lysed and their protein and mRNA expression analyzed by real-time PCR and western blot technique, respectively. SV2A, B proteins were identified in non-tumor (MCF-10A) and tumor cell lines (MDA-MB-231 and T47D) while SV2C only was found in the T47D cell line. Furthermore, the genomic expression was consistent with protein expression for a such cell line, but in MDA-MB-231 there was no SV2B genomic expression, and the SV2C mRNA and protein were not found in the non tumoral cell line. These findings suggest a possible cellular transdifferentiation to neural character in breast cancer, of possible relevance to cancer development, and point to possible use of SV2 as molecular marker and a vehicle for cancer treatment with botulinum toxin.

• The Plant Pathology Journal
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• 제31권2호
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• pp.108-114
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• 2015

Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.

• Journal of Microbiology and Biotechnology
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• 제28권8호
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• pp.1352-1359
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• 2018

Lactobacillus plantarum is a lactic acid bacterium that promotes animal intestinal health as a probiotic and is found in a wide variety of habitats. Here, we investigated the genomic features of different clusters of L. plantarum strains via pan-genomic analysis. We compared the genomes of 108 L. plantarum strains that were available from the NCBI GenBank database. These genomes were 2.9-3.7 Mbp in size and 44-45% in G+C content. A total of 8,847 orthologs were collected, and 1,709 genes were identified to be shared as core genes by all the strains analyzed. On the basis of SNPs from the core genes, 108 strains were clustered into five major groups (G1-G5) that are different from previous reports and are not clearly associated with habitats. Analysis of group-specific enriched or depleted genes revealed that G1 and G2 were rich in genes for carbohydrate utilization (${\text\tiny{L}}-arabinose$, ${\text\tiny{L}}-rhamnose$, and fructooligosaccharides) and that G3, G4, and G5 possessed more genes for the restriction-modification system and MazEF toxin-antitoxin. These results indicate that there are critical differences in gene content and survival strategies among genetically clustered L. plantarum strains, regardless of habitats.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.8-8
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• 2019

The stringent response (SR) is characterized as a bacterial defense mechanism in response to various growth-inhibiting stresses. It is activated by accumulation of a small nucleotide regulator, (p)ppGpp, and induces global changes in bacterial transcription and translation. Recent work from our group has shown that (p)ppGpp plays a critical role in virulence and survival in Vibrio cholerae. The genes, relA and relV, are involved in the production of (p)ppGpp, while the spoT gene encodes an enzyme that hydrolyzes it in V. cholerae. A mutant strain defective in (p)ppGpp production (i.e. ${\Delta}relA{\Delta}relV{\Delta}spoT$ mutant) lost the ability to produce cholera toxin (CT) and lost their viability due to uncontrolled production of organic acids, when grown with extra glucose. In contrast, the ${\Delta}relA{\Delta}spoT$ mutant, a (p)ppGpp overproducer strain, produced enhanced level of CT and exhibited better growth in glucose supplemented media via glucose metabolic switch from organic fermentation to acetoin, a neutral fermentation end product, fermentation. These findings indicates that (p)ppGpp, in addition to its well-known role as a SR mediator, positively regulates CT production and maintenance of growth fitness in V. cholerae. This implicates SR as a promising drug target, inhibition of which may possibly downregulate V. cholerae virulence and survival fitness. Therefore, we screened a chemical library and identified a compound that induces medium acidification (termed iMAC) and thereby loss of wild type V. cholerae viability under glucose-rich conditions. Further, we present a potential mechanism by which the compound inhibits (p)ppGpp accumulation. Together, these results indicate that iMAC treatment causes V. cholerae cells to produce significantly less (p)ppGpp, an important regulator of the bacterial virulence and survival response, and further suggesting that it has a therapeutic potential to be developed as a novel antibacterial agent against cholera.

• 한국식품영양학회지
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• 제33권6호
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• pp.702-709
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• 2020

The purpose of this study was to evaluate microbiological contamination of knives and cutting boards in child-care centers. Materials used in this study were swabbed of cutting boards and knives (blade, handle of knife, and joint of handle and blade) in 129 child-care centers. Mean values of total aerobic bacteria of swabs of knives and cutting boards were 1.7±0.7 log cfu/100 ㎠ and 1.7±0.9 log cfu/100 ㎠, respectively. Contamination levels of coliform bacteria from knives and cutting boards were 1.5±0.6 log cfu/100 ㎠ and 1.7±0.8 log cfu/100 ㎠, respectively. Comparing microbiological contamination levels of knives and cutting boards according to type and size of child-care centers, there was no significant difference. Bacillus cereus was detected in knife handles and cutting boards. Diarrhea-type toxin gene (entFM) was detected in B. cereus isolates. Antibiotic resistance tests showed that B. cereus was resistant to β-lactam antibiotics. To reduce microbiological contamination levels of knives and cutting boards in child-care centers and prevent food poisoning from bacteria contamination, continuous education by children's food-service management center is needed for sterilization and disinfection of knives and cutting boards.

• Journal of Microbiology and Biotechnology
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• 제33권3호
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• pp.329-338
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• 2023

Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that produces attaching and effacing lesions on the large intestine and causes hemorrhagic colitis. It is primarily transmitted through the consumption of contaminated meat or fresh produce. Similar to other bacterial pathogens, antibiotic resistance is of concern for EHEC. Furthermore, since the production of Shiga toxin by this pathogen is enhanced after antibiotic treatment, alternative agents that control EHEC are necessary. This study aimed to discover alternative treatments that target virulence factors and reduce EHEC toxicity. The locus of enterocyte effacement (LEE) is essential for EHEC attachment to host cells and virulence, and most of the LEE genes are positively regulated by the transcriptional regulator, Ler. GrlA protein, a transcriptional activator of ler, is thus a potential target for virulence inhibitors of EHEC. To identify the GrlA inhibitors, an in vivo high-throughput screening (HTS) system consisting of a GrlA-expressing plasmid and a reporter plasmid was constructed. Since the reporter luminescence gene was fused to the ler promoter, the bioluminescence would decrease if inhibitors affected the GrlA. By screening 8,201 compounds from the Korea Chemical Bank, we identified a novel GrlA inhibitor named Grlactin [3-[(2,4-dichlorophenoxy)methyl]-4-(3-methylbut-2-en-1-yl)-4,5-dihydro-1,2,4-oxadiazol-5-one], which suppresses the expression of LEE genes. Grlactin significantly diminished the adhesion of EHEC strain EDL933 to human epithelial cells without inhibiting bacterial growth. These findings suggest that the developed screening system was effective at identifying GrlA inhibitors, and Grlactin has potential for use as a novel anti-adhesion agent for EHEC while reducing the incidence of resistance.

• KSBB Journal
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• 제18권6호
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• pp.522-528
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• 2003

본 연구는 식중독 환자로부터 분리한 장염비브리오균 120 균주를 세균학적 특성시험, 항균제 감수성 여부, 독소 유전자와 독소조절유전자의 검출 및 보유 여부를 PCR, RPLA 로 시험하였으며, 각 지역간의 분리 세균을 GS-PCR, PFGE 시험으로 병원체 상관 관계를 분석한 결과 다음과 같은 결론을 얻었다. 장염비브리오균은 0%의 NaCl 농도에서는 성장을 보이지 않았고, 8% NaCl이 첨가된 농도에서는 성장을 보였다. 국내 설사환자의 장염비브리오균의 O, K 혈청형은 17가지로 나타났으며, 이 중 O3:K6형이 68.3%로 가장 많았다. Ampicillin 등 18가지 항균제 시험에서 Ampicillin, Ticacillin에 높은 내성을 보였으며, Ampicillin, Ticacillin, Vancomycin에 동시 내성을 나타낸 경우가 52.5%로 나타났다. 독소조절 유전자 toxR은 PCR시험에서 시험균주 모두 368bp 크기의 유전자를 가지고 있었으며 장염 비브리오균의 조기진단에 유용한 것으로 확인되었고, 독소유전자 tdh는 120균주 중 109 균주만이 199bp 크기의 유전자를 보유하고 11균주가 음성이었다. trh 독소 유전자를 보유한 균주는 시험균주 중 3 균주만이 250bp 크기의 유전자를 보유하고 있었으며, tdh, trh 유전자를 동시에 가지고 있는 균주는 3균주로 이 균주의 TDH 독소 생성능은 x16 정도로 독소 생성능이 미약했다. 독소생성 Kanagawa 시험에서 120균주 중 107균주가 양성반응 을 나타내었으며, Kanagawa 양성반응을 보인 균주은 모두 tdh 유전자를 보유하고 있었다. Group Specific-PCR에서 최근에 유행을 일으키는 O3:K6 혈청형의 유연관계를 찾는데 유용한 것으로 나타났다. 조절유전자 toxRS 염기서열을 분석한 결과 같은 혈청형이라도 3균주는 7개의 염기서열 차이를 나타내었으며, 이는 현재 유행을 일으키는 균주와는 다른 균주임을 확인하였다. PFGE로 보다 더 세분화하여 혈청관계를 분류할 수 있었으며, 식중독 환자 유래 장염비브리오균은 3가지 type으로 나타났으며, 이들 균주 사이에는 상호 밀접한 상관관계를 나타내었다. 따라서 PFGE 기법으로 얻어진 결과는 분자 역학적으로 유용한 도구로 사용될 수 있음을 확인하였다.

• 한국식품과학회지
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• 제47권1호
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• pp.126-131
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• 2015

본 연구에서는 한국의 전통적인 식품인 비빔밥에서 2005년에서 2011년에 분리한 대장균에 대한 특성을 연구하였다. 전체 1,142건의 비빔밥에서 84 균주(7.4%)의 대장균이 분리되었다. 분리한 균주의 항생제 내성 분석 및 다제 내성 패턴은 ampicillin의 내성이 가장 높았으며 6 균주(7.2%)에서는 다제 내성을 보였다. 분리한 균주의 병원성 유전자 13종의 유무를 확인한 결과 5 균주(5.9%)에서 ent 유전자를 보유하여 STEC/EPEC 형의 대장균으로 분류되었다. 그러나 EIEC, EAEC, ETEC 형은 검출되지 않았다. 병원성 유전자가 검출되지 않은 대장균에 대한 혈청형은 O103 (1.2%), O91 (1.2%), O128 (6.0%)으로 확인되었고 O26, O157, O145, O111, O121 혈청형은 없었다. 동일한 O 혈청형 균주는 독소 유전자를 가지고 있는 것과 없는 것이 동시에 있다고 보고 되어 있고 혈청형으로 병원성 유전자를 구분하는 것은 균주의 특성에 따라 차이가 많이 나므로 향후 연구가 필요하다. 위해 요소가 있는 균주에 대해서 유전적 상동성을 시험한 결과 균주의 상동성이 낮아서 서로 다른 오염원에서 분리 되었음을 유추 할 수 있었다. 비빔밥은 다양한 재료를 사용하며 제조 과정이 복잡하기 때문에 대장균에 의한 오염의 가능성이 매우 높은 식품이다. 그러므로 원재료부터 완성까지 위생적으로 철저하게 조리해야 하며 교차 오염 방지 및 관리를 통해 안심하고 섭취할 수 있도록 해야 한다.