• Title/Summary/Keyword: NF1-A

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A Cell-Based Assay System for Monitoring NF-$\kappa$B Activity in Human Epidermal Keratinocytes: A Screening Tool of the Antioxidants and Anti-inflammatories for Dermatological Purpose

  • Moon, Ki-Young;Hahn, Bum-Soo;Lee, Jinseon;Kim, Yeong-Shik
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.1
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    • pp.17-27
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    • 2001
  • A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.

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Pro-inflammatory Cytokine Expression Through NF-${\kappa}B/I{\kappa}B$ Pathway in Lung Epithelial Cells (폐 상피세포에서 NF-${\kappa}B/I{\kappa}B$ 경로에 의한 염증매개 사이토카인의 발현)

  • Park, Gye-Young;Lee, Seung-Hee;HwangBo, Bin;Yim, Jae-Joon;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.332-342
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    • 2000
  • Background : The importance of pro-inflammatory cytokines, especially tumor necrosis factor $\alpha$ (INF-$\alpha$) and interleukin-1$\beta$ (IL-1$\beta$), have been extensively documented in the generation of inflammatory lung disease. Lung epithelial cells are also actively involved in initiating and maintaining inflammation by producing pro-inflammatory mediators. Understanding the mechanism of pro-inflammatory cytokine expression in lung epithelial cells is crucial to the development of new therapeutic modalities for inflammatory lung disease. Transcription of most pro-inflammatory cytokines is dependent on the activation of NF-${\kappa}B$. However, the relationship between pro-inflammatory cytokine expression and NF-${\kappa}B/I{\kappa}B$ pathway in lung epithelial cells is not clear. Methods : BEAS-2B, A549, Na-H157, NCI-H719 cells were stimulated with IL-$1{\beta}$ or TNF-$\alpha$ at various times, and then IL-8 and TNF-$\alpha$mRNA expressions were assayed by Northern blot analysis. IL-$1{\beta}$ or TNF-$\alpha$-induced NF-${\kappa}B$ activation was assessed by the nuclear translocation of p65 NF-${\kappa}B$ subunit. The degradation of $I{\kappa}B{\alpha}$ and $I{\kappa}B{\beta}$ by IL-$1{\beta}$ or TNF-$\alpha$stimulation was assayed by Western blot analysis. The phosphorylation of $I{\kappa}B{\alpha}$ was evaluated by Western blot analysis after pre-treating cells with proteasome inhibitor followed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation. The basal level of IKK $\alpha$ expression was evaluated by Western blot analysis. Results: $I{\kappa}B{\alpha}$ and $I{\kappa}B{\alpha}$ was rapidly degraded after 5 minutes of incubation with IL-$1{\beta}$ or TNF-$\alpha$ in BEAS-2B, A549, and NCI-H157 cells. The activation of NF-${\kappa}B{\alpha}$ and the induction of IL-8 and TNF-$\alpha$ mRNA expression were observed by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in these cells. In contrast, neither the changes in NF-${\kappa}B/I{\kappa}B$ pathway nor IL-8 and TNF-$\alpha$mRNA expression was induced by IL-$1{\beta}$ or TNF-$\alpha$ stimulation in NCI-H719 cells. IL-$1{\beta}$ and TNF-$\alpha$-induced $I{\kappa}B$ phosphorylation was observed in BEAS-2B, A549, and NCI-H157 cells, but not in NCI-H719 cells. The basal level of IKK$\alpha$ expression was not different between cell. Conclusion : NF-${\kappa}B/I{\kappa}B$ pathway plays an important role in the expression of pro-inflammatory cytokine in most lung epithelial cells. The absence of the effect on NF-${\kappa}B/I{\kappa}B$ pathway in NCI-H719 cells sæms to be due to the defect in the intracellular signal transduction pathway upstream to IKK.

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Fertilizer Effect of Waste Nutrient Solution in Greenhouses for Young Radish Cultivation (열무 재배를 위한 시설하우스 폐양액의 비료 효과)

  • Hong, Youngsin;Moon, Jongpil;Park, Minjung;Son, Jinkwan;Yun, Sungwook
    • Journal of Bio-Environment Control
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    • v.31 no.4
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    • pp.460-467
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    • 2022
  • The purpose of this study is to enhance utilization of the waste nutrient solution (WNS) disposed at the hydroponic greenhouse. Several sets of testing were conducted to examine the effects of WNS: (a) a fertilizer effect, (b) soil column leaching, and (c) crop cultivation. The fertilizer effect test was applied in young radish cultivation by examining the growth characteristics of young radish and soil based on inorganic nitrogen according to the soil treatment of the nitrogen fertilizer (NF) and the WNS. The fertilizer effects and crop cultivation test were conducted with five treatments (A-E): A, non-treatment (water); B, 100% of NF; C, 70% of NF + 30% of WNS; D, 50% of NF + 50% of WNS; and E, 30% of NF + 70% of WNS. The soil column leaching test was conducted with three treatments: non-treatment (water), 100% of NF, 50% of WNS + 50% of NF. As a result, the chemical properties of the WNS were pH 6.0, EC 2.4dS·m-1, total phosphorus (T-P) 28mg·L-1, ammonium nitrogen (NH4-N) 5.0mg·L-1, and nitrate nitrogen (NO3-N) 301mg·L-1. The chemical properties of the soil were pH 5.51, EC 0.31dS/m, organic matter 2.08g·kg-1, NO3-N 9.64mg·kg-1, and NH4-N 3.20mg·kg-1. The results of fertilizer effects showed that the ratio of 50% or less of NF and 50% or more of WNS was high in young radish growth. There was no statistically significant difference between the soil chemistry in the C-E treatments where WNS was mixed with NF and the B treatment where only NF was applied. As a result of the soil column leaching test, there was no significant difference in the concentrations of NO3 and NH4 in the treatment of 100% of NF and 50% of NF + 50% of WNS. The study indicates, if the mixed fertilizer of WNS and NF is applied in the soil cultivation of young radish, it will reduce the use of NF and environmental pollution. This also helps reduce production costs on farmers and increase the yield of young radish.

Solid-phase Synthesis and Preliminary Evaluation of 1,6,8-Trisubstituted Tetrahydro-2H-pyrazino[1,2-a]pyrimidine-4,7-diones as a NF-kB Inhibitor

  • Kim, Jin-Woong;Cho, Jung-Hyuck;Han, Tae-hee;Lee, Jong-Baek;Oh, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.27 no.4
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    • pp.484-488
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    • 2006
  • The solid-phase synthesis of new series of 1,6,8-trisubstituted tetrahydro-2H-pyrazino[1,2-a]pyrimidine-4,7-diones as bicyclic $\beta$-turn mimetics is described. Their NF-kB inhibition activities were tested and the effect of substituents on bicyclic ring was investigated. Among the prepared compounds, the fluorobenzyl and methoxybenzyl group substituted compounds 26 and 27 at C-1 and C-8 position showed more inhibitory activities than the others. Tested at a concentration of 10 uM, these two compounds showed a 60% inhibition against the target NF kB 549.

Expression of Nuclear Factor Kappa B (NF-κB) as a Predictor of Poor Pathologic Response to Chemotherapy in Patients with Locally Advanced Breast Cancer

  • Prajoko, Yan Wisnu;Aryandono, Teguh
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.595-598
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    • 2014
  • Background: NF-${\kappa}B$ inhibits apoptosis through induction of antiapoptotic proteins and suppression of proapoptotic genes. Various chemotherapy agents induce NF-${\kappa}B$ translocation and target gene activation. We conducted the present study to assess the predictive value of NF-${\kappa}B$ regarding pathologic responses after receiving neoadjuvant chemotherapy. Materials and Methods: We enrolled 131 patients with locally advanced invasive ductal breast carcinoma. Immunohistochemistry (IHC) was used to detect NF-${\kappa}B$ expression. Evaluation of pathologic response was elaborated with the Ribero classification. Results: Expression of NF-${\kappa}B$ was significantly associated with poor pathological response (p=0.02). From the multivariate analysis, it was found that the positive expression of NF-${\kappa}B$ yielded RR=1.74 (95%CI 0.77 to 3.94). Conclusions: NF-${\kappa}B$ can be used as a predictor of poor pathological response after neoadjuvant chemotherapy.

Wideband Resistive LNA based on Noise-Cancellation Technique Achieving Minimum NF of 1.6 dB for 40MHz (40MHz에서 1.6 dB 최소잡음지수를 얻는 잡음소거 기술에 근거한 광대역 저항성 LNA)

  • Choi Goangseog
    • Journal of Korea Society of Digital Industry and Information Management
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    • v.20 no.2
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    • pp.63-74
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    • 2024
  • This Paper presents a resistive wideband fully differential low-noise amplifier (LNA) designed using a noise-cancellation technique for TV tuner applications. The front-end of the LNA employs a cascode common-gate (CG) configuration, and cross-coupled local feedback is employed between the CG and common-source (CS) stages. The moderate gain at the source of the cascode transistor in the CS stage is utilized to boost the transconductance of the cascode CG stage. This produces higher gain and lower noise figure (NF) than a conventional LNA with inductor. The NF can be further optimized by adjusting the local open-loop gain, thereby distributing the power consumption among the transistors and resistors. Finally, an optimized DC gain is obtained by designing the output resistive network. The proposed LNA, designed in SK Hynix 180 nm CMOS, exhibits improved linearity with a voltage gain of 10.7 dB, and minimum NF of 1.6-1.9 dB over a signal bandwidth of 40 MHz to 1 GHz.

Evaluation of Physicochemical Properties of Jeungpyun Prepared with Membrane-filtered Tofu Whey Concentrates

  • Joo, Sin-Youn;Choi, Min-Hee;Jung, Jin-Young;Kim, Woo-Jung;Chung, Hai-Jung
    • Food Quality and Culture
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    • v.2 no.1
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    • pp.43-47
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    • 2008
  • This study examined the quality characteristics of Jeungpyun prepared with different additions of nano-filtered (NF) tofu whey concentrates. The initial pH values of the Jeungpyun batters ranged from 5.64 to 5.78, and decreased to 4.77-4.98 after 4 hours of fermentation at $35^{\circ}C$. The volume and specific volume values of the control Jeungpyun were lower than those of Jeungpyun samples prepared with 1%, 2%, and 3% NF powder. The color of the Jeungpyun became increasingly greenish-yellow as the NF powder level increased. Hardness and brittleness decreased with increasing NF powder content, while cohesiveness and springiness were not significantly different. Sensory evaluations revealed that as the level of NF powder increased, takju smell and sourness increased, but no significant differences were observed for sweetness and moistness between the control and NF powder groups. In terms of overall acceptability, the results revealed that Jeungpyun can be prepared with up to 1% NF powder in place of rice powder and be deemed as acceptable as a control Jeungpyun product.

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ZAS3 promotes TNFα-induced apoptosis by blocking NFκB-activated expression of the anti-apoptotic genes TRAF1 and TRAF2

  • Shin, Dong-Hyeon;Park, Kye-Won;Wu, Lai-Chu;Hong, Joung-Woo
    • BMB Reports
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    • v.44 no.4
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    • pp.267-272
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    • 2011
  • ZAS3 is a large zinc finger transcription repressor that binds the ${\kappa}B$-motif via two signature domains of ZASN and ZASC. A loss-of-function study showed that lack of ZAS3 protein induced accelerated cell proliferation and tumorigenesis. Conversely, gain-of-function studies showed that ZAS3 repressed $NF{\kappa}B$-activated transcription by competing with $NF{\kappa}B$ for the ${\kappa}B$-motif. Based on these observations, we hypothesize that ZAS3 promotes apoptosis by interrupting anti-apoptotic activity of $NF{\kappa}B$. Here, we present evidence that upon $TNF{\alpha}$ stimulation, ZAS3 inhibits $NF{\kappa}B$-mediated cell survival and promotes caspase-mediated apoptosis. The inhibitory effect of ZAS3 on $NF{\kappa}B$ activity is mediated by neither direct association with $NF{\kappa}B$ nor disrupting nuclear localization of $NF{\kappa}B$. Instead, ZAS3 repressed the expression of two key anti-apoptotic genes of $NF{\kappa}B$, TRAF1 and TRAF2, thereby sensitizing cells to $TNF{\alpha}$-induced cell death. Taken together, our data suggest that ZAS3 is a tumor suppressor gene and therefore serves as a novel therapeutic target for developing anti-cancer drugs.

TAK1-dependent Activation of AP-1 and c-Jun N-terminal Kinase by Receptor Activator of NF-κB

  • Lee, Soo-Woong;Han, Sang-In;Kim, Hong-Hee;Lee, Zang-Hee
    • BMB Reports
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    • v.35 no.4
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    • pp.371-376
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    • 2002
  • The receptor activator of nuclear factor kappa B (RANK) is a member of the tumor necrosis factor (TNF) receptor superfamily. It plays a critical role in osteoclast differentiaion, lymph node organogenesis, and mammary gland development. The stimulation of RANK causes the activation of transcription factors NF-${\kappa}B$ and activator protein 1 (AP1), and the mitogen activated protein kinase (MAPK) c-Jun N-terminal kinase (JNK). In the signal transduction of RANK, the recruitment of the adaptor molecules, TNF receptor-associated factors (TRAFs), is and initial cytoplasmic event. Recently, the association of the MAPK kinase kinase, transforming growth factor-$\beta$-activated kinase 1 (TAK1), with TRAF6 was shown to mediate the IL-1 signaling to NF-${\kappa}B$ and JNK. We investigated whether or not TAK1 plays a role in RANK signaling. A dominant-negative form of TAK1 was discovered to abolish the RANK-induced activation of AP1 and JNK. The AP1 activation by TRAF2, TRAF5, and TRAF6 was also greatly suppressed by the dominant-negative TAK1. the inhibitory effect of the TAK1 mutant on RANK-and TRAF-induced NF-${\kappa}B$ activation was also observed, but less efficiently. Our findings indicate that TAK1 is involved in the MAPK cascade and NF-${\kappa}B$ pathway that is activated by RANK.

Vitamin C Blocks TNF-${\alpha}$-induced NF-kB Activation and ICAM-1 Expression in Human Neuroblastoma Cells

  • Son, Eun-Wha;Mo, Sung-Ji;Rhee, Dong-Kwon;Pyo, Suhk-Neung
    • Archives of Pharmacal Research
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    • v.27 no.10
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    • pp.1073-1079
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    • 2004
  • Interactions of the cell adhesion molecules are known to play important roles in mediating inflammation. The proinflammatory cytokine, tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), activates the NF-kB signaling pathway, which induces the expression of various genes, such as intercellular adhesion molecule-1 (ICAM-1). In this study, the effect of vitamin C on the ICAM-1 expression induced by TNF-${\alpha}$ in a human neuroblastoma cell line, SK-N-SH was investigated. Treatment with vitamin C resulted in the downregulation of the TNF-${\alpha}$-induced surface expression and ICAM-1 mRNA levels in a concentration-dependent manner. Moreover, a gel shift analysis indicated that vitamin C dose-dependently inhibited the NF-kB activation and IkB${\alpha}$ degradation induced by TNF-${\alpha}$. Taken together, these results suggest that vitamin C downregulates TNF-${\alpha}$- induced ICAM-1 expression via the inhibition of NF-kB activation.