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Zika Virus-Encoded NS2A and NS4A Strongly Downregulate NF-κB Promoter Activity

  • Lee, Jeong Yoon;Nguyen, Thi Thuy Ngan;Myoung, Jinjong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1651-1658
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    • 2020
  • Since Zika virus (ZIKV) was first detected in Uganda in 1947, serious outbreaks have occurred globally in Yap Island, French Polynesia and Brazil. Even though the number of infections and spread of ZIKV have risen sharply, the pathogenesis and replication mechanisms of ZIKV have not been well studied. ZIKV, a recently highlighted Flavivirus, is a mosquito-borne emerging virus causing microcephaly and the Guillain-Barre syndrome in fetuses and adults, respectively. ZIKV polyprotein consists of three structural proteins named C, prM and E and seven nonstructural proteins named NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5 in an 11-kb single-stranded positive sense RNA genome. The function of individual ZIKV genes on the host innate immune response has barely been studied. In this study, we investigated the modulations of the NF-κB promoter activity induced by the MDA5/RIG-I signaling pathway. According to our results, two nonstructural proteins, NS2A and NS4A, dramatically suppressed the NF-κB promoter activity by inhibiting signaling factors involved in the MDA5/RIG-I signaling pathway. Interestingly, NS2A suppressed all components of MDA5/RIG-I signaling pathway, but NS4A inhibited most signaling molecules, except IKKε and IRF3-5D. In addition, both NS2A and NS4A downregulated MDA5-induced NF-κB promoter activity in a dosedependent manner. Taken together, our results suggest that NS2A and NS4A signifcantly antagonize MDA5/RIG-I-mediated NF-κB production, and these proteins seem to be controlled by different mechanisms. This study could help understand the mechanisms of how ZIKV controls innate immune responses and may also assist in the development of ZIKV-specific therapeutics.

Gene Expression Profiling of Human Bronchial Epithelial (BEAS-2B) Cells Treated with Nitrofurantoin, a Pulmonary Toxicant

  • Kim, Youn-Jung;Song, Mee;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.222-230
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    • 2007
  • Some drugs may be limited in their clinical application due to their propensity towards their adverse effects. Toxicogenomic technology represents a useful approach for evaluating the toxic properties of new drug candidates early in the drug discovery process. Nitrofurantoin (NF) is clinical chemotherapeutic agent and antimicrobial and used to treatment of urinary tract infections. However, NF has been shown to result in pulmonary toxic effects. In this research, we revealed the changing expression gene profiles in BEAS-2B, human bronchial epithelial cell line, exposed to NF by using human oligonucleotide chip. Through the clustering analysis of gene expression profiles, we identified 136 up-regulated genes and 379 down-regulated genes changed by more than 2-fold by NF. This study identifies several interesting targets and functions in relation to NF-induced toxicity through a gene ontology analysis method including biological process, cellular components, molecular function and KEGG pathway.

Effect of Reaction Gases on PFCs Treatment Using Arc Plasma Process (아크 플라즈마를 이용한 과불화합물 처리공정에서 반응가스에 의한 효과)

  • Park, Hyun-Woo;Choi, Sooseok;Park, Dong-Wha
    • Clean Technology
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    • v.19 no.2
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    • pp.113-120
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    • 2013
  • The treatment of chemically stable perflourocompounds (PFCs) requires a large amount of energy. An energy efficient arc plasma system has been developed to overcome such disadvantage. $CF_4$, $SF_6$ and $NF_3$ were injected into the plasma torch directly, and net plasma power was estimated from the measurement of thermal efficiency of the system. Effects of net plasma power, waste gas flow rate and additive gases on the destruction and removal efficiency (DRE) of PFCs were examined. The calculation of thermodynamic equilibrium composition was also conducted to compare with experimental results. The average thermal efficiency was ranged from 60 to 66% with increasing waste gas flow rate, while DRE of PFCs was decreased with increasing gas flow rate. On the other hand, DRE of each PFCs was increased with the increasing input power. Maximum DREs of $CF_4$, $SF_6$ and $NF_3$ were 4%, 15% and 90%, respectively, without reaction gas at the fixed input power and waste gas flow rate of 3 kW and 70 L/min. A rapid increase of DRE was found using hydrogen or oxygen additional gases. Hydrogen was more effective than oxygen to decompose PFCs and to control by-products. The major by-product in the arc plasma process with hydrogen was hydrofluoric acid that is easy to be removed by a wet scrubber. DREs of $CF_4$, $SF_6$ and $NF_3$ were 25%, 39% and 99%, respectively, using hydrogen additional gas at the waste gas flow rate of 100 L/min and the input power of 3 kW.

Therapeutic Effects of Yijungtang on Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse Induced by Mite Antigen (이중탕(理中湯)이 Mite Antigen으로 유발된 NC/Nga 생쥐의 아토피 피부염에 미치는 영향)

  • Seo, Hui-Yeon;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.25 no.1
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    • pp.1-27
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    • 2011
  • Objectives: The purpose of this study is to investigate the effects of Yijungtang(YJT) on atopic dermatitis in an in-vitro and in-vivo experiment using a RBL-2H3 mast cells and a NC/Nga atopic dermatitis mouse. Methods: In-vitro experiment, IL-4, IL-13 mRNA expression were evaluated by a real-time PCR, IL-4, IL-13 production by ELISA and transcription factor as GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-kB by western blotting. In-vivo experiment, clinical skin score we evaluated by, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mouse, cytokine level, total number of cell, Immunohistochemical staining and Histological features of auxiliary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue in NC/Nga mouse. Results: YJT decreased IL-4, IL-13 mRNA expression, IL-4, IL-13 production and prominently decreased the expression of mast cell specific transcription factors including GATA-2, NF-AT2, c-Fos and NF-kB. YJT oral administration reduced the levels of skin severity scores. It also decreased the level of inflammatory cytokines such as IL-5, IL-13, histamine and IgE in the serum. It elevated IFN-gamma level in the spleenocyte culture supernatant but decreased. $CD3e^+$, $CD19^+$, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $CD11b^+Gr-1^+$, $CCR3^+$ in the PBMCs, $CD4^+$, $CD8^+$, $CD3e^+CD69^+$, $B220^+CD23^+$ in the ALN, $CD4^+$, $CD3e^+CD69^+$ in the ALN and $CD4^+$, $CD11b^+Gr-1^+$ in the dorsal skin. Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mice were much improved by YJT oral administration. Conclusions: The anti-allergic activities of YJT may be mediated by down-regulation of Th2 cytokines, such as IL-4 and IL-13, through the regulation GATA-2, NF-AT2 and NF-kB transcription factors in mast cells. YJT would be regulate molecular mediators and immune cells which are functionally associated with atopic dermatitis induced in NC/Nga mice, and may play an important role in recovering AD symptoms.

Purification and Characterization of a Thermostable Xylanase from Paenibacillus sp. NF1 and its Application in Xylooligosaccharides Production

  • Zheng, Hong-Chen;Sun, Ming-Zhe;Meng, Ling-Cai;Pei, Hai-Sheng;Zhang, Xiu-Qing;Yan, Zheng;Zeng, Wen-Hui;Zhang, Jing-Sheng;Hu, Jin-Rong;Lu, Fu-Ping;Sun, Jun-She
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.489-496
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    • 2014
  • High levels of extracellular xylanase activity (211.79 IU/mg) produced by Paenibacillus sp. NF1 were detected when it was submerged-cultured. After three consecutive purification steps using Octyl-Sepharose, Sephadex G75, and Q-Sepharose columns, a thermostable xylanase (XynNF) was purified to homogeneity and showed a molecular mass of 37 kDa according to SDS-PAGE. The specific activity of the purified XynNF was up to 3,081.05 IU/mg with a 14.55-fold purification. The activity of XynNF was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Fe^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $Cu^{2+}$, SDS, and EDTA. The purified XynNF displayed a greater affinity for oat spelt xylan with the maximal enzymatic activity at $60^{\circ}C$ and pH 6.0. XynNF, which was shown to be cellulose-free, with high stability at high temperature ($70^{\circ}C-80^{\circ}C$) and low pH range (pH 4.0-7.0), is potentially valuable for various industrial applications. The enzyme hydrolyzed oat spelt xylan to yield mainly xylooligosaccharides (95.8%) of 2-4 degree of polymerization (DP2-4). Moreover, the majority of the xylooligosacharides (DP2-4) products was xylobiose (61.5%). The thermostable xylanase (XynNF) thus seems potentially usefull in the production of xylooligosaccharides.

Treatment of AP Solutions Extracted from Solid Propellant by NF/RO Membrane Process (NF/RO 멤브레인 공정을 적용한 고체추진제에서 추출된 암모늄 퍼클로레이트 (AP) 처리 연구)

  • Kong, Choongsik;Heo, Jiyong;Yoon, Yeomin;Han, Jonghun;Her, Namguk
    • Membrane Journal
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    • v.22 no.4
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    • pp.235-242
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    • 2012
  • Ammonium perchlorate (AP) is primarily derived from the process of liquid incineration treatment when dismantling a solid rocket propellant. A series of batch dead-end nanofiltration (NF) and reverse osmosis (RO) membrane experiments were conducted to explore the retention mechanisms of AP under various hydrodynamic and solution conditions. Low levels of silicate type of siloxane had been detected through the GC/MS and FTIR analysis of liquid solutions extracted from solid ammonium perchlorate composite propellant (APCP). It is indicated that NF/RO membranes fouling in the presence of APCP was mainly attributed to the AP interactions because the concentration of silicate type of siloxane was negligible compared to that of AP. The osmotic pressure of AP was presumably resulted in the flux declines ranging from 13 to 17% in the case of the application of low-pressure (551 and 896 kPa for NF and RO) compared to those in application of high-pressure. The retention of AP by NF/RO membranes significantly varied from approximately 10 to 70% for NF and 26 to 87% for RO, depending on the operating and solution water chemistry conditions. The results suggested that retention efficiency of AP was fairly increased by reducing concentration polarization (i.e. application of low-pressure and stirring speed of 600 rpm) and increasing the pH of a solution. The result of this study was also consistent with the previous modeling of 'solute mass transfer of NF/RO membranes' and demonstrated that hydrodynamic and solution water chemistry conditions are to be a key factor in the retention of AP by NF/RO membranes.

The Effects of Sunbanghwalmyung-eum Extract on Anti-oxidant, Anti-allergic and Anti-inflammatory ability in mouse cell (Mouse cell에서 선방활명음(仙方活命飮)의 항산화작용과 항알러지 및 항염증 효과)

  • Park, Min-Chul;Hong, Seung-Ug
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.21 no.2
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    • pp.46-53
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    • 2008
  • Background and Objectives : The aim of this study was to investigate the anti-oxidant, anti-allergic and anti-inflammatory ability of the Taglisodog-eum(SHE) extract on the RAW 264.7 and EL4 cells Materials and Methods : Three types of experiments were implemented for this study: first, the experiment to study the anti-oxidant effect of SHE using Riboflavin; second, in vitro experiment to investigate the inhibition of Th 2 cell differentiation by SHE using EL4 cells (IL-4 mRNA expression); third, the suppression of $NF-{\kappa}B$ activation using RAW 264.7 cells (iNOS and COX-2 mRNA expression). Results : The anti-oxidant ability of SHE were dose-dependantly increased. From in vitro, the LPS-induced iNOS and COX-2 mRNA expression were dose-dependantly decreased in the RAW264.7 cells treated with SHE and the PMA-induced IL-4 mRNA expression were also dose-dependantly decreased in EL4 cells. $NF-{\kappa}B$ activation was suppressed, and iNOS & COX-2 production were inhibited by SHE Conclusion : The results suggest that SHE has dose-dependant anti-oxidant ability, and has anti-allergic and anti-inflammatory effects through the suppression of $NF-{\kappa}B$ activation and the inhibition of Th 2 cell differentiation.

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Involvement of TLR4-JNK/NF-κB signaling pathway in RAW264.7 cell activation of Protaetia brevitarsis seulensis larvae extracts (흰점박이꽃무지 유충 추출물의 RAW264.7 세포 활성화에서 TLR4-JNK/NF-κB 신호전달 경로의 관여)

  • Ju-Hwi Park;Jongbeom Chae;Joon Ha Lee;Dongyup Hahn;Ju-Ock Nam
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.447-454
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    • 2023
  • In the environment in which humans live, there are various antigens that invade the human body and interfere with humans leading a healthy life, so the immune system recognizes the antigen then removes them through a complex mechanism. Macrophages are widely distributed immune cells involved in the innate immune system, and produce various immune modulators such as inducible nitric oxide synthase-induced nitric oxide, cyclooxygenase-2 induced prostaglandin E2 and proinflammatory cytokines such as tumor necrosis factor-alpha. On the other hand, Protaetia brevitarsis seulensis larvae are a type of edible insect that have emerged as an alternative to the future food supply problem. The immuno-modulatory effect through the activation of murine macrophage RAW264.7 cell via mitogen-activated protein kinases (MAPKs)/nuclear factor-kappa B (NF-κB) signaling pathways has been reported. Based on this report, in this study, we confirmed how the expression of immune modulators induced by Protaetia brevitarsis seulensis larvae extracts in RAW264.7 cells was changed by treatment with pharmacological inhibitors of toll-like receptor 4 (TLR4), MAPKs and NF-κB signaling pathways. As a result, reduction of immune modulators was confirmed in the c-Jun N-terminal kinase (JNK) inhibitor treatment group and NF-κB inhibitor treatment group among the Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell. Furthermore, in the TLR4 inhibitor-treated group, decreases in phosphorylation of JNK and NF-κB factors were confirmed in Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell, as well as decreases in immune modulators. This results suggest that Protaetia brevitarsis seulensis larvae activates RAW264.7 cells by the engagement of TLR4-JNK/NF-κB signaling pathway.

Effect of Bcl-2 on Apoptosis and Transcription Factor NF-κB Activation Induced by Adriamycin in Bladder Carcinoma BIU87 Cells

  • Zhang, Guo-Jun;Zhang, Zhe
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2387-2391
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    • 2013
  • Resistance to apoptosis is a major obstacle preventing effective therapy for malignancies. Bcl-2 plays a significant role in inhibiting apoptosis. We reconstructed a stable human Bcl-2 transfected cell line, BIU87-Bcl-2, that was derived from the transfection of human bladder carcinoma cell line BIU87 with a plasmid vector containing recombinant Bcl-2 [pcDNA3.1(+)-Bcl-2]. A cell line transfected with the plasmid alone [pcDNA3.1(+)-neo] was also established as a control. BIU87 and BIU87-neo proved sensitive to adriamycin induced apoptosis, while BIU87-Bcl-2 was more resistant. In view of the growing evidence that NF-${\kappa}B$ may play an important role in regulating apoptosis, we determined whether Bcl-2 could modulate the activity of NF-${\kappa}B$ in bladder carcinoma cells. Stimulation of BIU87, BIU87-neo and BIU87-Bcl-2 with ADR resulted in an increase expression of NF-${\kappa}B$ (p<0.001). The expression of NF-${\kappa}B$ in BIU87-Bcl-2 was higher than in the other two cases, with a concomitant reduction in the $I{\kappa}B{\kappa}$ protein level. These results suggest that the overexpression of Bcl-2 renders human bladder carcinoma cells resistant to adriamycin-induced cytotoxicity and there is a link between Bcl-2 and the NF-${\kappa}B$ signaling pathway in the suppression of apoptosis.

Anti-inflammatory Effects of Flavokavain C from Kava (Piper methysticum) Root in the LPS-induced Macrophages (LPS로 유도된 대식세포에서 카바뿌리로부터 분리한 Flavokavain C의 항염증 효과)

  • Park, Chung;Han, Jong-Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.311-320
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    • 2016
  • Kava (Piper methysticum, P. methysticum) is used as traditional herbal medicine for urogenital diseases, rheumatisms, gastrointestinal problems, respiratory irritations, and pulmonary pains. We identified a flavokavain C (FKC) from P. methysticum, which showed anti-inflammatory activity on nuclear factor ${\kappa}B$ (NF-${\kappa}B$)-dependent nitric oxide (NO) production and expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. FKC inhibited accumulation of reactive oxygen species (ROS), such as hydrogen peroxide, and was able to dose-dependently reduce the LPS-induced NO production and the expression of various inflammation-associated genes (iNOS, IL-$1{\beta}$, IL-6) through inhibition of NF-${\kappa}B$ and MAPKs (ERK and JNK). In conclusion, these results indicate that FKC may have the potential to prevent inflammation process including NF-${\kappa}B$ and MAPKs pathways, and it could be applicable to functional cosmetics for anti-inflammation and antioxidant properties.