• Title/Summary/Keyword: RT Component

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Particulate Matter from Asian Dust Storms Induces the Expression of Proinflammatory Cytokine in A549 Epithelial Cells (PM10이 A549 Cells에서 전염증성 Cytokine발현에 미치는 영향)

  • Kim, Jung Ho;Jeon, Hyo Keun;Kim, Mi Kyeong;Kyung, Sun Yong;An, Chang Hyeok;Lee, Sang Pyo;Park, Jung Woong;Jeong, Sung Hwan
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.6
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    • pp.663-672
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    • 2006
  • Background: $PM_{10}$(Particulate matter with a diameter ($<10{\mu}m$), which is characterized by different environmental conditions, is a complex mixture of organic and inorganic compounds. The Asian dust event caused by meteorological phenomena can also produce unique particulate matter in affected areas. This study investigated the cytokine produced by A549 epithelial cells exposed to particles collected during both the Asian dust pfenomenon and ambient air particles in a non-dusty period. Method: Air samples were collected using a high volume air sampler(Sibata Model HV500F) with an air flow at $500{\ell}/min$ for at least 6 hours. The cytokine messenger RNA(mRNA) was measured using a reverse transcriptase polymerase chain reaction(RT-PCR). The A549 cells were exposed to 10 to $500{\mu}g/m{\ell}$ of a suspension containing $PM_{10}$ for 24 hours. Each was compared with those in the non-exposed control cells. Result: The mRNA levels of interleukin(IL)-$1{\alpha}$, $IL-I{\beta}$, IL-8, and the granulocyte macrophage colony stimulating factor(GM-CSF) increased after veing exposed to $PM_{10}$ in the ambient air particles, compared with those in the non-exposed control cells. The increase in $IL-1{\alpha}$ and IL-8 were dose dependent at a $PM_{10}$ concentration between $100{\mu}g/m{\ell}$ and $500{\mu}g/m{\ell}$. The mRNA level of IL-8 in the A549 epithelial cells was higher during the in the Asian dust period($500{\mu}g/m{\ell}$) than during the non dust period. Conclusion: A549 cells exposed to the $PM_{10}$ collected during the Asian dust period produce more proinflammatory cytokine than during non-dusty period. This cytokine enhances the local inflammatory response in the airways and can also contribute to the systemic component of this inflammatory process.

The Effect of Kaempferol, guercetin on Hyaluronan-Synthesis Stimulation in Human Keratinocytes (HaCaT) (인체 피부 세포주 (HaCaT)에서 Kaempferol, Quercetin의 Hyaluronan 합성 촉진 효과에 대한 연구)

  • Kim, Seung-Hun;Nam, Gae-Won;Kang, Byung-Young;Lee, Hae-Kwang;Moon, Seong-Joon;Chang, Ih-Seop
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.97-102
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    • 2005
  • One of the key molecules involved in skin moisture is hyaluronan (hyaluronic acid, HA) with its associated water of hydration. The predominant component of the ECM (extracellular matrix) of skin is HA. It Is the primordial and the simplest of the GAGs (glycosaminoglycans), a water-sorbed macromolecule In extracellular matrix, Included between the vital cells of epidermis. In the skin, HA was previously thought to derive extlusively from dermis. But, recent studies revealed that HA could be synthesized in epidermis. Flavonoids are polyphenolic compounds that is found mainly in foods of plant origin. Kaempferol was known to increase glutathione synthesis in human keratinocyte. And quercetin blocked PPAR-meidated keratinocyte differentiation as lipoxygenase inhibitors. In this study, we sought to evaluate the effect of flavonid, kaempferol and quercetin on production HA in keratinocyte. We examined the changes of three human hyaluronan synthase genes (HASI, HAS2, HAS3) expression by semi-quantitative RT-PCR when kaempferol or quercetin was added to cultured human keratinocytes. We found that these flavonoids slightly upregulated HAS2, HAS3 mRNA after 24 h. And we investigated the effect on HA production by ELISA. When we evaluated the level of HA in culture medium after 24 h incubation. We found enhanced accumulation of HA in the culture medium. Although the effects of above flavonoids are less than retinoic acid, the data indicate that kaempferol, quercetin can dose-dependently increase the level of HA in epidermis cell line. It suggested that flavonoid, kaempferol, and quercetin increased production of HA in skin and it helped to elevate skin moisture and improve facial wrinkle.

Convergence Studies of NO Homeostasis in Cellular Signalling (세포의 신호전달 과정에서 NO 항상성에 관한 융복합 연구)

  • Oh, Hee-Kyun;Do, Eun-Young;Park, Hae-Ryoung
    • Journal of Digital Convergence
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    • v.13 no.12
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    • pp.461-467
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    • 2015
  • Saussurea lappa is known for a variety of physiological activities as a component but has not known to show the effect of the cellular signaling pathway. We investigated the anti-inflammatory effects by Saussurea lappa ethanol extract on the LPS(lipopolysaccharide) induced nitric oxide (NO) production by RAW 264.7 cell line. It shows the expressions of iNOS and COX-2 at the transcriptional level (RT-PCR). The Saussurea lappa ethanol extract showed transcriptional expression levels of pro-inflammatory cytokine TNF- and IL-$1{\beta}$ induced by LPS(lipolysaccharide) in RAW264.7 cell line. Saussurea lappa ethanol extract reduced the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the RNA levels in a concentration-dependent manner. The finding that ethanol extract of Saussurea lappa has an influence on NO (nitric oxide) homeostasis in the study of the action mechanism on the macrophage-mediated inflammatory reaction was considered in terms of convergence. And it is to provide an important basis for the prevention and treatment of inflammatory diseases in the future.

Chitosan Increases α6 Integrinhigh/CD71high Human Keratinocyte Transit-Amplifying Cell Population

  • Shin, Dong-Wook;Shim, Joong-Hyun;Kim, Yoon-Kyung;Son, Eui-Dong;Yang, Seung-Ha;Jeong, Hye-Jin;Lee, Seok-Yong;Kim, Han-Kon;Park, Soo-Nam;Noh, Min-Soo
    • Biomolecules & Therapeutics
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    • v.18 no.3
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    • pp.280-285
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    • 2010
  • Glycosaminoglycans (GAGs) and chitosan have been used as matrix materials to support the dermal part of skin equivalent which is used for both pharmacological and toxicological evaluations of drugs potentially used for dermatological diseases. However, their biological roles of GAGs and chitosan in the skin equivalent are still unknown. In the present study, we evaluated whether GAGs and chitosan directly affect keratinocyte stem cells (KSCs) and their transit-amplifying cells (TA cells). Among supporting matrix materials, chitosan significantly increased the number of ${\alpha}6$ $integrin^{high}/CD71^{high}$ human keratinocyte TA cells by 48.5%. In quantitative real-time RT-PCR analysis, chitosan significantly increased CD71 and CD200 gene transcription whereas not ${\alpha}6$ integrin. In addition, the level of the gene transcription of both keratin 1 (K1) and K10 in the chitosan-treated human keratinocytes was significantly lower than those of control, suggesting that chitosan inhibit keratinocyte differentiation. We also found that N-acetyl-D-glucosamine (NAG) and $\beta$-(1-4)-linked D-glucosamine (D-glc), two components of chitosan, have no effect on the expression of CD71, K1, and K10, suggesting that each monomer component of chitosan is not enough to regulate the number of epidermal keratinocyte lineage. Conclusively, chitosan increases keratinocyte TA cell population which may contribute to the cellular mass expansion of the epidermal part of a skin equivalent system.

Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

Prevalence and Molecular Characterization of Quinolone Antibiotic Resistance in Escherichia coli Isolates from Raw Bulk Milk in Gyeonggi-do (원유시료에서 분리한 대장균의 퀴놀론 항생제 내성 기전)

  • Kang, Sowon;Lee, Sangjin;Choi, Sungsook
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.185-190
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    • 2014
  • The aim of this study was to investigate the prevalence of quinolone resistant E. coli from raw bulk milk and to characterize the resistance determinants. In this study, the gyrA, gyrB, parC, and parE quinolone resistance determining regions (QRDR) were sequenced from quinolone resistant E. coli isolates. Also, the presence of plasmid-mediated quinolone resistance (PMQR) and the expression of efflux pump genes based on quantitative real-time PCR (qRT-PCR) were investigated. Of the 487 coliform bacteria, 9 strains showed nalidixic acid resistance, and 6 of the 9 nalidixic acid resistant isolates were also ciprofloxacin resistant. These 9 strains had a single mutation at codon 83 (S83L) in gyrA, 2 of them had double mutations at codon 83 and 87 (S83L and D87N) in gyrA and 3 of the 9 isolates had single mutations at codon 80 (S80I) in parC. None of the 9 isolates harbored PMQR determinants. Compared with wild-type E. coli ATCC 25922, an over-expression of the acrB gene (2.15-5.74 fold), encoding the pump component of the AcrAB-TolC efflux pump was observed in 4 of 6 ciprofloxacin resistant isolates. This study identified the quinolone resistance mechanism of E. coli isolated from raw milk samples in Gyeonggi-do.

A Study on the Torsional Vibration Characteristics of Super Large Two Stroke Low Speed Diesel Engines with Tuning Damper (튜닝댐퍼를 갖는 초대형 저속 2행정 디젤엔진의 비틀림진동 특성에 관한 연구)

  • Lee, Don-Chool;Barro, Ronald D.
    • Transactions of the Korean Society for Noise and Vibration Engineering
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    • v.19 no.1
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    • pp.64-75
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    • 2009
  • The shipbuilder's requirement for a higher power output rating has led to the development of a super large two stroke low speed diesel engines. Usually a large-sized bore engine ranging from $8{\sim}14$ cylinders, this engine group is capable of delivering power output of more than 100,000 bhp at maximum continuous rating(mcr). Other positive aspects of this engine type include higher thermal efficiency, reliability, durability and mobility. This plays a vital role in meeting the propulsion requirement of vessels, specifically for large container ships, of which speed is a primary concern to become more competitive. Consequently, this also resulted in the modification of engine parameters and new component designs to meet the consequential higher mean effective pressure and higher maximum combustion pressure. Even though the fundamental excitation mechanisms unchanged, torsional vibration stresses in the propulsion shafting are subsequently perceived to be higher. As such, one important viewpoint in the initial engine design is the resulting vibration characteristic expected to prevail on the propulsion shafting system(PSS). This paper investigated the torsional vibration characteristics of these super large engines. For the two node torsional vibration with a nodal point on the crankshaft, a tuning damper is necessary to reduce the torsional stresses on the crankshaft. Hence, the tuning torsional vibration damper design and compatibility to the shafting system was similarly reviewed and analyzed.

Cloning and Structural Analysis of bfmo Operon in Methylophaga aminosulfidovorans SK1 (Methylophaga aminosulfidovorans SKI bfmo 오페론의 클로닝 및 구조 분석)

  • Lim Hyun Sook;Goo Jae Whan;Kim Lee Hyun;Kim Si Wouk;Cho Eun Hee
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.1-7
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    • 2005
  • Methylophaga aminosulfidovorans SK1 (KCTC 10323 BP) can utilize trimethylamine as a sole carbon, nitrogen, and energy source. The bacterial flavin-containing monooxygenase (bFMO) gene was identified in the strain and the recombinant enzyme expressed in E. coli oxidized trimethylamine. To study the function and regulation of the bfmo, over 8,000 nucleotide sequences of the neighboring regions including the bfmo were determined. Three open reading frames proceeding to the bfmo gene encoded analogues to highly conserved nitrate/nitrite sensing two-component system regulators and a methyl accepting protein. Two small open reading frames just downstream of the bfmo gene showed no similar proteins of known functions but the sequences were conserved among other bacteria. Reverse transcription-polymerase chain reaction analysis showed that the six putative genes consisted of three transcription units. The three regulatory genes located upstream of the bfmo gene formed two separate transcription units. The bfmo and the two downstream genes were transcribed from a single promoter.

Physiochemical Properties of Functional Oils Produced Using Red Yeast-Rice Ethanol Extracts and Diacylglycerol Oil (홍국쌀 에탄올 추출물과 Diacylglycerol Oil을 이용하여 제조한 기능성 유지의 이화학적 특성 연구)

  • Kim, Nam-Sook;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.2
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    • pp.201-208
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    • 2007
  • Functional oils (FOs) were produced from commercial diacylglycerol oil and red yeast rice extracts from 80% ethanol for 1 hr in a shaking water bath at $35^{\circ}C$ and 175 rpm. FOs contained (A) 600, (B) 1200, (C) 1800, and (D) 2280 ppm of red yeast-rice extracts, respectively. The Hunter a value and b value were risen whereas L value was reduced along with the increase of extract concentration. Content of monacolin K and total phenolic compounds in FOs significantly increased according to the increase of extract concentration. The oxidation stability of FOs was observed by Rancimat at $98^{\circ}C$. Induction time decreased according to the increase of extract concentration. The major volatile compounds of FOs were compared using the electronic nose (EN) system and solid phase microextraction (SPME) method combined with gas chromatograph/mass spectrometry (GC/MS). EN was composed of 12 different metal oxide sensors. Sensitivities (Rgas/Rair) of sensors from EN were analyzed by principal component analysis (PCA), whose proportion was 99.66%. For qualitative or quantitative analysis of volatile compounds by SPME-GC/MS, the divinylbenzene/carboxene/polydimethyl-siloxane fiber and sampling temperature of $50^{\circ}C$ were applied.

Mechanism underlying NO-induced apoptosis in human gingival fibroblasts

  • Hwang, In-Nam;Jeong, Yeon-Jin;Jung, Ji-Yeon;Lee, Jin-Ha;Kim, Kang-Moon;Kim, Won-Jae
    • International Journal of Oral Biology
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    • v.34 no.1
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    • pp.7-14
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    • 2009
  • Nitric oxide (NO) acts as an intracellular messenger at the physiological level but can be cytotoxic at high concentrations. The cells within periodontal tissues, such as gingival and periodontal fibroblasts, contain nitric oxide syntheses and produce high concentrations of NO when exposed to bacterial lipopolysaccharides and cytokines. However, the cellular mechanisms underlying NO-induced cytotoxicity in periodontal tissues are unclear at present. In our current study, we examined the NO-induced cytotoxic mechanisms in human gingival fibroblasts (HGF). Cell viability and the levels of reactive oxygen species (ROS) were determined using a MTT assay and a fluorescent spectrometer, respectively. The morphological changes in the cells were examined by Diff-Quick staining. Expression of the Bcl-2 family and Fas was determined by RT-PCR or western blotting. The activity of caspase-3, -8 and -9 was assessed using a spectrophotometer. Sodium nitroprusside (SNP), a NO donor, decreased the cell viability of the HGF cells in a dose- and time-dependent manner. SNP enhanced the production of ROS, which was ameliorated by NAC, a free radical scavenger. ODQ, a soluble guanylate cyclase inhibitor, did not block the SNP-induced decrease in cell viability. SNP also caused apoptotic morphological changes, including cell shrinkage, chromatin condensation, and DNA fragmentation. The expression of Bax, a member of the proapoptotic Bcl-2 family, was upregulated in the SNP-treated HGF cells, whereas the expression of Bcl-2, a member of the anti-apoptotic Bcl-2 family, was downregulated. SNP augmented the release of cytochrome c from the mitochondria into the cytosol and enhanced the activity of caspase-8, -9, and -3. SNP also upregulated Fas, a component of the death receptor assembly. These results suggest that NO induces apoptosis in human gingival fibroblast via ROS and the Bcl-2 family through both mitochondrial- and death receptor-mediated pathways. Our data also indicate that the cyclic GMP pathway is not involved in NO-induced apoptosis.