• Tuberculosis and Respiratory Diseases
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• v.58 no.6
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• pp.590-599
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• 2005

Object : Cigarette smoking is a major cause of mucus hypersecretion, which is a pathophysiological feature of many inflammatory airway diseases. Mucins, which are an important part of the airway mucus, are synthesized from the Muc gene in airway epithelial cells. However, the signaling pathways for cigarette smoke-induced mucin synthesis are unknown. The aim of this study was to determine the signal pathway for smoking induced Muc5ac gene expression. Methods : A549 cells were cultured and transiently transfected with the Muc5ac promoter fragment. These cells were stimulated with 5% cigarette smoke extract (CSE) alone or with CSE after a pretreatment with various signal transduction pathway inhibitors (AG1478, PD98059 and SB203580). The Muc5ac promoter activity was examined using the luciferase reporter system, and the level of phosphorylated EGFR, ERK1/2, p38 MAPK and JNK were all examined using Western blot analysis. Muc5ac mRNA expression was also examined using reverse transcriptase polymerase chain reactions (RT-PCR). Results : 1. The peak level of luciferase activity of the Muc5ac promoter was observed at 5% concentration and after 3 hours of incubation with the CSE. The level of EGFR phosphorylation and the luciferase activity of the transfected cells caused by the CSE were significantly suppressed by AG1478 or PD98059 (P<0.01). 2. CSE phosphorylated ERK1/2 or p38 MAPK but not JNK. The Muc5ac mRNA expression level was increased by the CSE but that was suppressed by PD98059 or AG1478. 3. The CSE-induced phosphorylation of ERK1/2 was blocked by PD98059 and that of p38 MAPK was blocked by either PD98059 or SB203580. Either PD98059 or SB203580 suppressed the luciferase activity of the transfected cells (P<0.0001). Conclusion : The Muc5ac mRNA expression level was increased by the CSE. The increased CSE-induced transcriptional activity was mediated via EGF receptor activation, which led to ERK1/2 and p38 MAPK phosphorylation.

• Applied Chemistry for Engineering
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• v.5 no.3
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• pp.501-508
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• 1994

1-Ethyl-6-fluoro-1, 4-dihydro-4-oxo-7-(1-piperazinyl)quinolinea-3-car-boxylic acid-dextran was synthesized by the reaction of 1-ethyl-6-fluoro-1, 4-dihydro-4-oxo-7-(1-piperazinyl )quinoline-3-acryloyl chloride with dextran. Polymeric drug was tested for antimicrobial activity in vitro against ten species of microorganisms. Polymeric drug revealed good antibacterial activity against Bacillus subtillis ATCC 6633, Staphyloccus aureus ATCC 25923, Mycrobactertum phlei IFO 3158, Salmonella typhimurium KCTC 1925, Escherichia coli KCTC 1039, Escherichia coli ESS, Klebsiella puemouiae KCTC 1560 and Pseudomonas aeruginosa IFO 13130. Polymeric drug have no antimicrobial against Candida albicans ATCC 10231, but moderately active Micrococcus luteus ATCC 9341.

• The Journal of Natural Sciences
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• v.14 no.1
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• pp.51-61
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• 2004

T7 RNA polymerase is a single subunit RNA polymerase able to accomplish whole transcription process without auxiliary factors. T7 phage lysozyme involcing in destruction of host cell wall repress T7 transcription and affects transcription termination process. Therefore expression vector pT7lys containing T7 phage lysozyme gene was constructed and expressed. T7 phage lysozyme protein was purified to homogeneity by Ni-NTA column chromatography. Also amidase activity of the purified lysozyme was identified. In order to understand the effect of the lysozyme on the sequence specific transcription termination. T7 transcription elongation complexes at the site rrnB T1 transcription termination signal were made in the presence the lysozyme. The results shows that the transcription elongation complexes are unstable in the presence of T7 phage lysozyme.

• The Korean Journal of Pesticide Science
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• v.9 no.4
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• pp.429-436
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• 2005

As Sclerotinia sclerotiorum causing cucumber sclerotinia rot was the fastest in the mycelial growth at $25^{\circ}C$, its pathogenicity was strong at the same temperature among several temperatures. All the isolates of Sclerotinia sclerotiorum showed a strong pathogenicity against cucumber fruits, which was confirmed by a disk assay and a wound assay. A wound assay was superior to a disk assay to develop the assay system for assessing the fungicidal activity of several fungicides against Sclerotinia sclerotiorum. In a disk assay, it was very difficult to assess the fungicidal activity, because the pathogenicity of isolates used in the experiment was very strong. At 500 and $3.0{\mu}g/mL$, the activity of dichloflouanid and the mixture of carbendazim and diethofencarb against cucumber sclerotinia rot was 14.3 and 42.3%, respectively, by using a disk assay. However, at same concentration two fungicides showed the high controlling activity as 100 and 92.5%, through a wound assay in a laboratory. Also, the activity of two fungicides was good against cucumber sclerotinia rot in the greenhouse where cucumber plants were cultivated in the field, showing the control value as 91.1 and 82.9% at 100 and $825{\mu}g/mL$, respectively. All the isolates of Sclerotinia sclerotiorum from cucumber fruits sampled in the polyvinyl house were subjected to monitoring for the resistance to 7 fungicides. The $EC_{50}$ value of 7 fungicides was as follows: fenhexamid; $0.13{\mu}g/mL$, procymidon and iprodione; 0.18 and $0.24{\mu}g/mL$, carbendazim and the mixture of carbendazim and diethofencarb; 0.13과 $0.05{\mu}g/mL$, iminoctadine and dichlofluanid; 1.94 and $8.95{\mu}g/mL$. Ultimately it was not found that resistant isolates of Sclerotinia sclerotiorum were appeared in the field.

• Microbiology and Biotechnology Letters
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• v.43 no.4
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• pp.357-366
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• 2015

Quorum sensing (QS) is involved in the process of cell-to-cell communication and as a gene regulatory mechanism, which has been implicated in bacterial pathogenicity. Bacteria use this QS system to control a variety of physiological processes. In this study, pomegranate (Punica granatum L.) peel extract (PPE) was first screened for its ability to inhibit QS in bio-reporter strains (Chromobacterium violaceum and C. violaceum CV026). Next, the ability of PPE to inhibit swimming motility and biofilm formation was examined in Y. enterocolitica. Additionally, changes in the expression of specific genes involved in the synthesis of the N-acylhomoserine lactones (AHLs; yenI and yenR) and in the flagellar regulon (fliA, fleB and flhDC) were evaluated by reverse transcription (RT)-PCR. The results show that PPE specifically inhibited and reduced QS-controlled violacein production by 78.5% in C. violaceum CV026, and decreased QS-associated biofilm formation and swimming motility in Y. enterocolitica without significantly affecting bacterial growth. These inhibitory effects were also associated with the down-regulation of gene expression involved in the synthesis of AHLs and in motility. Our results suggest that PPE could be a potential therapeutic agent to prevent enteropathogens in humans, as well as highlight the need to further investigate the in vivo properties of PPE for clinical applications.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.12
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• pp.1701-1707
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• 2012

3,3'-Diindolylmethane (DIM) is a major in vivo derivative of the putative anticancer agent indole-3-carbinol, which is present in cruciferous vegetables and has been reported to have anti-carcinogenic properties. An abnorrmally elevated level of cyclooxygenase-2 (COX-2) has been implicated in the pathogenesis of carcinogenesis. To investigate the mechanism by which DIM exhibits anti-carcinogenic effects, we investigated the effects of DIM on COX-2 expression in MCF-10A human mammary epithelial cells treated with the tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA). DIM inhibited TPA-induced COX-2 expression and suppressed the synthesis of prostaglandin $E_2$, one of the major products of COX-2. Nuclear factor-kappa B ($NF-{\kappa}B$) is a transcription factor known to play a role in regulation of COX-2 expression. Treatment of MCF-10A cells with TPA increased nuclear translocation of phospho-p65, with the maximal levels being reached at 1 hour, while DIM inhibited the TPA-induced nuclear translocation of phospho-p65. Overall, we demonstrated that DIM suppresses phorbol ester-induced $PGE_2$ production and COX-2 expression in MCF-10A cells. The reduction in COX-2 levels by DIM maybe mediated through inhibition of $NF-{\kappa}B$ signaling.

• The Korean Journal of Food And Nutrition
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• v.11 no.3
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• pp.272-277
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• 1998

We carried out separation and guantitation of flavor components by GC about essential oils extracted from deodorized corn oil at the different deodorizing temperature. Flavor components were detected total 16 kinds included aldehydes of 8 kinds, major components were propane, pentane, hexanal etc. These major components content was about 70~75% of the total flavor components. According to rise of deodorizing temperature, both ethane and aldehydes of 8 kinds content were in proportion to increase, but propane, pentane, hexane, octan, pentyl furan content were decreased by contraries, respectively. On the other hand, total flavor component content was appeared the lowest level at 245$^{\circ}C$ treating group, aldehydes content was in proportion to increase according to rise of deodorizing temperature. These phenomenons consider that the undesirable reactions such as partial auto-oxidation, degradation, polymerization and hydrolysis etc. by effecting factors of stripping steam and vacuum degree. Conclusively, deodorizing temperature under high temperature was undesirable for the minimization of off-flavor materials.

• Tuberculosis and Respiratory Diseases
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• v.44 no.4
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• pp.796-805
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• 1997

Background : It is clear that deregulation of cell cycle progression is a hallmark of neoplastic transformation and genes involved in the $G_1$/S transition of the cell cycle are especially frequent targets for mutations in human cancers, including lung cancer. p16 gene product, one of the G1 cell-cycle related proteins, that is recently identified plays an important role in the negative regulation of the the kinase activity of the cyclin dependent kinase (cdk) enzymes. Therefore p16 gene is known to be an important tumor suppressor gene and is also called MTS1 (multiple tumor suppressor 1). No more oncogenes have been reported to be frequently related to multiple different malignancies than the alterations of p16 gene. Especially when it comes to non-small cell lung cancer, there was no expression of p16 in more than 70% of cell lines examined. And also it is speculated that p16 gene could exert a key role in the development of non-small cell lung cancer. This study was designed to evaluate whether p16 gene could be used as a candidate for gene therapy of non-small cell lung cancer. Methods : After the extraction of total RNA from normal fibroblast cell line and subsequent reverse transcriptase reaction and polymerase chain reaction, the amplified p16 cDNA was subcloned into eukaryotic expression plasmid vector, pRC-CMV. The constructed pRC-CMV-p16 was transfected into the NCI-H441 NSCLC cell line using lipofectin. The changes of G1 cell-cycle related proteins were investigated with Western blot analysis and immunoprecipitation after extraction of proteins from cell lysates and tumor suppressive effect was observed by clonogenic assay. Results : (1) p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 showed the formation of p16 : cdk 4 complex and decreased phosphorylated Rb protein, while control cell line did not. (2) Clonogenic assay demonstrated that the number of colony formation was markedly decreased in p16(-) NCI-H441 cell line transfected with pRC-CMV-p16 than the control cell line. Conclusion : It is confirmed that the expression of p16 protein in p16 absent NSCLC cell line with the gene transfection leads to p16 : cdk4 complex formation, subsequent decrease of phosphorylated pRb protein and ultimately tumor suppressive effects. And also it provides the foundation for the application of p16 gene as a important candidate for the gene therapy of NSCLC.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.1
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• pp.15-24
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• 2003

The purpose of this study is to investigate the sterilization effect of Er:YAG laser against the intraoral acid producing bacterium, S. mutans, by irradiating the culture solution containing S. mutans KCTC 3065 with Er:YAG laser having a $650{\mu}m$ diameter beam through the non-contact method. We obtained the following results after examining the temperature changes of the culture solution, numbers of bacterial colonies, and acid-producing ability and attaching ability on teeth by measuring the amount of extracellular polysaccharide produced by S. mutans. The number of bacterial colony was decreased in $10{\mu}l$ culture solution irradiated with laser in overall compared to the control solution. The number decreased as the irradiation intensity and pulse repetition rate were larger and as the exposure time was increased. However, it did not change significantly in $100{\mu}l$ culture solution compared to the control solution. Although the acid-producing ability of S. mutans was inhibited for a certain duration after laser irradiation in 10r1 bacterial culture solution, it did not change in $100{\mu}m$ solution compared with the control solution. The amount of extracellular polysaccharide synthesized by S. mutans was partially decreased through laser irradiation in $10{\mu}m$ culture solution but did not change in $100{\mu}m$ culture solution. Based on these findings, we concluded that Er:YAG laser has an sterilization effect on S. mutans in which we presume that the mechanism is through the heat effect rather than the mechanical effect from development of ultrasound.

• Microbiology and Biotechnology Letters
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• v.39 no.2
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• pp.104-110
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• 2011

Actinomycetes, Gram positive soil bacteria, are valuable microorganisms which produce useful secondary metabolites including antibiotics, antiparasitic substances, anti-cancer drugs, and immunosuppressants. Although a major family of actinomycetes, known as streptomycetes, has been intensively investigated at the molecular level for several decades, a potentially valuable and only recently isolated non-streptomycetes rare actinomycetes (NSRA) family has been poorly characterized due to lack of proper genetic manipulation systems. Here we report that a PCR-based genome screening strategy was performed with approximately 180 independently isolated actinomycetes strains to isolate potentially valuable NSRA strains. Thanks to this simple PCR-based genome screening strategy we were able to identify only seven NSRA strains, followed by 16S rRNA sequencing for confirmation. Through further bioassays, one potentially valuable NSRA strain (tentatively named Nocardiopsis species MMBL010) was identified which possessed both antifungal and antibacterial activities, along with the presence of polyketide synthase and non-ribosomal peptide synthase genes. Moreover, Nocardiopsis species MMBL010, which was intrinsically recalcitrant to genetic manipulation, was successfully transformed via E. coli-driven conjugation. These results suggest that PCR-based genome screening, followed by the establishment of an E. coli-driven conjugation system, is an efficient strategy to maximize potentially valuable compounds and their biosynthetic genes from NSRA strains isolated from various environments.