• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.6
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• pp.481-489
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• 2010

Introduction: TLR-5, a member of the toll-like receptor (TLR) family, is a element of the type I transmembrane receptors, which are characterized by an intracellular signaling domain homolog to the interleukin-1 receptor. These receptors recognize microbial components, particularly bacterial flagellin. All-trans retinoic acid (atRA, tretinoin), a natural metabolite of vitamin A, acts as a growth and differentiation factor in many tissues, and is also needed for immune functions. In this study, THP-1 human macrophage-monocytes were used to examine the mechanisms by which atRA regulated the expression of TLR-5. Because the molecular mechanism underlying this regulation at the transcriptional level is also unclear, this study examined which putative transcription factors are responsible for TLR-5 expression by atRA in immune cells. Materials and Methods: This study examined whether atRA induces the expression of TLR-5 in THP-1 cells using reverse transcription-polymerase chain reaction (RT-PCR), and which transcription factors are involved in regulating the TLR-5 promoter in RAW264.7 cells using a reporter assay system. Western blot analysis was used to determine which signal pathway is involved in the expression of TLR-5 in atRA-treated THP-1 cells. Results: atRA at a concentration of 10 nM greatly induced the expression of TLR-5 in THP-1 cells. Human TLR-5 promoter contains three Sp-1/GC binding sites around -50 bp and two NF-kB binding sites at -380 bp and -160 bp from the transcriptional start site of the TLR-5 gene. Sp-1/GC is primarily responsible for the constitutive TLR-5 expression, and may also contribute to NF-kB at -160 bp to induce TLR-5 after atRA stimulation in THP-1 cells. The role of NF-kB in TLR-5 expression was further confirmed by inhibitor pyrrolidine dithiocarbamate (PDTC) experiments, which greatly reduced the TLR-5 transcription by 70-80%. Conclusion: atRA induces the expression of the human TLR-5 gene and NF-kB is a critical transcription factor for the atRA-induced expression of TLR-5. Accordingly, it is conceivable that retinoids are required for adequate innate and adaptive immune responses to agents of infectious diseases. atRA and various synthetic retinoids have been used therapeutically in human diseases, such as leukemia and other cancers due to the antiproliferative and apoptosis inducing effects of retinoids. Therefore, understanding the molecular regulatory mechanism of TLR-5 may assist in the design of alternative strategies for the treatment of infectious diseases, leukemia and cancers.

• Biomedical Science Letters
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• v.4 no.1
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• pp.43-56
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• 1998

A multiplex PCR method was designed by employing primers specific for the eaeA gene, conserved sequences of Shiga-like toxins (SLT-I.II), and the 60-MDa plasmid of enterohemorrhagic E. coli (EHEC) O157:H7 strain. A set of six synthetic oligonucleotide primers derived from sequences of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 were used in a multiplex PCR amplification procedure to detect these genes in the same enteric pathogens. In two enterohemorrhagic E. coli O157:H7 (ATCC 35150, ATCC 43894) reference strains, PCR products of 317bps (eaeA), 228bps (SLT-I.II), and 167bps (60-MDa plasmid) were successfully amplified simultaneously in a single reaction. However, the specific PCR products were not amplified in control strains of other enteric bacteria. The sensitivity of the multiplex PCR assay for detection of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 was found to be 2.5$\times$10$^{6}$ of bacteria in diarrheal stool to amplify all three bands. The multiplex PCR technology will allow large-scale screening of many clinical specimens or contaminated foods, and will be a very useful method for the detection of a wide range of microorganisms present in the environment, including EHEC O157:H7 in various types of specimens. The multiplex PCR assay has the potential to be used as a specific and rapid method for clinical diagnosis of disease caused by EHEC O157:H7.

• Journal of Periodontal and Implant Science
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• v.24 no.3
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• pp.472-482
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• 1994

Cytokines expressed specifically in leukocytes subsets and in activated cells, which are involved in chemotaxis and activation of leukocytes, are recently defined as chemokines. Macrophage inflammatory $protein-1{\alpha}(MIP-1{\alpha})$ and $MIP-1{\beta}$ are members of C-C chemokine subfamily which produces wide immunomodulatory, proinflammatory, and hematopoietic modulatory actions. We have studied their gene expression by using Northern blot analysis in various blood cells such as cytolytic T lymphocyte(CTL), helper T lymphocyte(HTL), macrophage, and B lymphocyte. Resting CTL line CTLL-R8 expressed $MIP-1{\alpha}$ mRNA which was downregulated by ConA stimulation. Both of resting and ConA stimulated HTL line Hut78 and Jurkat did not express $MIP-1{\alpha}$ mRNA. There was detectable $MIP-1{\alpha}$ transcript in HTL hybridoma 2B4.11 which was a little upstimulated by ConA stimulation. B cell line 230, and macrophage cell line RAW264.7 and WR19M.1 showed distinct $MIP-1{\alpha}$ message which were induced after LPS stimulation. Expression pattern of $MIP-1{\beta}$ in all cell lines or cell were almost identical to that of $MIP-1{\alpha}$. Also strategies employed to identify and characterize the biological functions was preceded by receptor cloning to trace the shorcut to the final goal of cytokine research. For the cloning of $MIP-1{\alpha}$ receptor(R), we used synthetic oligonucleotides of transmembrane(T) conserved sequences of already cloned human(h) IL-8-R, and performed reverse transcription-polymerase chain reaction(RT-PCR) amplification using murine(m) macrophage cell line mRNA. Among 5RT-PCR products, we isolated a homologous cDNA with hIL-8-R which were shown to be putative mIL-8-R cDNA.

• Journal of Bio-Environment Control
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• v.25 no.4
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• pp.240-248
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• 2016

The objective of this study was to determine the effect of exogenous abscisic acid (ABA) on the red coloration and endogenous ABA contents of apple fruit skins. ABA and fluridone (an ABA synthetic inhibitor, FD) was sprayed on 'Hongro' apple fruit skins at 107 days after full bloom (DAFB). Visual coloration and hunter's color values were not affected by the ABA and FD treatments. Anthocyanin contents in fruit skins increased similarly to hunter $a^*$ values of fruit skins, but ABA and FD did not affect its accumulations. Liquid chromatography analysis revealed that endogenous ABA contents in control fruit increased at first and then decreased from 12 hours after the treatment. ABA treatment increased ABA contents in fruit skins from 2 hour after the treatment and it lasted until the end of the treatments. FD decreased ABA contents in fruit skins from 6 hours after the treatment. ABA treatment increased MdNCED2 (an ABA biosynthetic gene), MdACO1 (an ethylene biosynthetic gene), and MdCHS and MdDFR expressions. However, MdUFGT expressions were not affected by ABA treatment.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.317-322
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• 2005

The cycloinulooligosaccharide fructanotransferase (CFTase) gene (cft) from Paenibacillus macerans was subcloned into an E. coli-yeast shuttle vector, pYES2.0, resulting in pYGECFTN. The plasmid pYGECFTN (8.6 kb) was introduced into Saccharomyces cerevisiae SEY2102 cells and then the transformants were selected on the synthetic defined media lacking uracil. The cft gene expression in yeast transformant was demonstrated by the analyses cyclofructan (CF) spots on thin-layer chromatogram. The recombinant CFTase was not secreted into the medium and localized in the periplasmic space. The production of CF was observed after 5 min of the enzymatic reaction with inulin. The optimun pH and temperature for CF production were found to be at pH 8.0 and $45^{\circ}C$, respectively. Enzyme activity was stably maintained up to $55^{\circ}C$. The CF was produced from all inulin sources and was most efficiently produced from dahlia tubers and Jerusalem artichokes.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.241-247
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• 2007

The cycloinulooligosaccharide fructanotransferase (CFTase) gene (cft) from Paenibacillus macerans was subcloned into the surface display vector, pCTcon (GAL1 promoter). The constructed plasmid, pCTECFTN (9.0 kb) was introduced to S. cerevisiae EBY100 cell and then east transformants were selected on the synthetic defined medium lacking uracil and on the inulin containing medium. The surface display of CFTase was confirmed by immunofluorescence microscopy and its enzymatic ability to form cycloinulooligosaccharides(cyclofructans, CFs) from inulin. The total activity of the CFTase was reached about 5.52 unit/1 by cultivation of yeast transformant on YPDG medium. The optimized conditions determined were as follows; pH, 8.0; temperature, $50^{\circ}C$ ; substrate concentration, 5%; inulin source, Jerusalem artichoke. By the reaction with inulin, CFs consisting of cycloinulohexaose (CF6), cycloinuloheptaose (CF7), and cycloinulooctaose (CF8) were produced and CF6 was the major product.

• Korean Journal of Agricultural Science
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• v.29 no.1
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• pp.32-43
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• 2002

The peach pyralid moth (Dichocrocis punctiferalis) is reported as import ant pest insect to chestnut fruits in Asia including Korea. For seasonal occurrence of D. punctiferalis in 1996, 1998 and 1999, D. punctiferalis adults were collected by light trap and sex pheromone wing trap in the chestnut orchard. Also, damaged rate of chestnut fruits were examined each collecting site. In 1996, according to examination by using light trap at three different site in Kongju, the occurrence peak of 2nd gene ration of D. punctiferalis adults was early August. Collected individuals after September were cons ide red as 3rd generation. Damaged rate due to maturation type was 26.5% averagely. The order of damaged rate were early, late and middle maturation type as 35.04%, 19.35% and 26.03%, respectively. Using synthetic sex pheromone trap at Kongju, Cheungyang and Buyeo in 1998, occurrence peaks were occurred mid-August to 2nd generation and late September to 3nd gene ration in Kongju. However, from early-August to late-September 5 peaks were occurred sequentially in Cheungyang. Second generation adults were occurred in mid-August and 3rd ones were occurred from late-September to early-October in Buyeo. In 1999, occurrence peaks of 3rd generation were early, middle and late September in Kongju, Cheungyang and Buyeo respectively. Damaged rate were 6.8%, 6.6% and 15.3% in Kongju, Cheungyang and Buyeo, respectively. Damaged rate order due to variety were Eunki, Chukpa, Danpa, Bokpa, Wangjung, Byunggo57, Yuoma at Buyeo and were Chukpa, Dukmyung, Byunggo57 at Cheungyang in 1999. Damage rate of chestnut were different with variety. Eunki and Chukpa in Buyeo and Chukpa and Dukmyung in Cheungyang were more damaged by D. punctiferalis respectively.

• Korean Journal of Organic Agriculture
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• v.22 no.4
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• pp.855-868
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• 2014

Synthetic agro-chemicals have been widely used to control diseases on paprika but these days negative attention has been increasing to use of them because of several adverse effects. This research was conducted to isolate and to characterize the antagonistic microorganism to control major paprika diseases, gray mold rot, fruit and stem rot, phytophthora blight, sclerotium rot, and wilt disease. Analysis of the fatty acid and analysis of the 16S rDNA gene sequence revealed that YKJ1 isolated in this research belongs to a group of Serratia marcescens. Specially, 16S rDNA gene sequence of YKJ1 showed 99% of sequence similarity with S. marcescens. Observation through the optical microscope revealed that YKJ1 suppressed the spore germination and the hyphal growth of pathogens. YKJ1 treatment on pathogens induced marked morphological changes like hyphal swelling and degradation of cell wall. In the case of phytophthora blight, the zoosporangium formation was restrained. S. marcescens found in this study call as S. marcescens-YKJ1 and it may be valuable as one of biological control agents against major diseases of paprika in the future even though it is require to be tested with more study on field test.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.98-102
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• 1996

A thermostable tyrosine phenol-lyase gene of a thermophilic Symbiobacterium species was cloned and overexpressed in Escherichia coli in order to produce the biocatalyst for the synthesis of 3, 4-dihy-droxyphenyl-L-alanine (L-DOPA). The substrates used for the synthetic reaction were pyrocatechol, so-dium pyruvate, and ammonium chloride. The enzyme was stable up to $60^{\circ}C$, and the optimal temperature for the synthesis of L-DOPA was $37^{\circ}C$ . The optimal pH of the reaction was about 8.3. Enzyme activity was highly dependent on the amount of ammonium chloride and the optimal concentration was estimated to be 0.6 M. In the case of pyrocatechol, an inactivation of enzyme activity was observed at con-centrations higher than 0.1 M. Enzyme activity was increased by the presence of ethanol. Under op-timized conditions, L-DOPA production was carried out adding pyrocatechol and sodium pyruvate to the reaction solution intermittently to avoid substrate depletion during the reaction. The concentration of L-DOPA reached 29.8 g/l after 6 h, but the concentration didn t increase further because of the formation of byproducts by a non-enzymatic reaction between L-DOPA and pyruvate.

• Korean Journal of Environmental Agriculture
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• v.29 no.1
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• pp.39-46
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• 2010

Environmental-friendly agriculture (EFA) is defined as the cultivation of crops with reduced amounts or without chemical-synthetic pesticides. Recently, the use of chemical pesticides has decreased significantly; therefore, we cultivated peppers following EFA- and conventional methods and compared their antioxidant activities. To accomplish this, the environmental-friendly cultivated peppers (EFPE) and conventionally cultivated peppers (CCPE) were extracted with 70% methanol and the effects of the extracts on the cell viability, intracellular ROS generation, lipid peroxidation and catalase activity of HepG2 cells were evaluated. EFPE showed a stronger protective effect against oxidative stress induced-cell death than that of CCPE. EFPE also reduced intracellular ROS generation (42.7% to 26.4%) following treatment with hydrogen peroxide more effectively than that of CCPE (24.2% to 6.3%). Furthermore, EFPE and CCPE showed protective effects against lipid peroxidation and induced catalase activity, although these effects were not statistically significant. Taken together, these results suggest that EFPE showed stronger antioxidant activities than CCPE, and thus represent evidence that EFA with biocontrol materials may improve the functional properties of crops and/or secondary metabolites with antioxidant activities when compared with conventional agricultural practices.