• Development and Reproduction
• /
• v.15 no.1
• /
• pp.25-30
• /
• 2011

Embryonic stem (ES) cells can self-renew maintaining the undifferentiated state. Self renewal requires many factors such as Oct4, Sox2, FoxD3, and Nanog. NF-${\kappa}B$ is a transcription factor involved in many biological activities. Expression and activity of NF-${\kappa}B$ increase upon differentiation of ES cells. Reportedly, Nanog protein directly binds to NF-${\kappa}B$ protein and inhibits its activity in ES cells. Here, we found a potential binding site of NF-${\kappa}B$ in the human Nanog promoter and postulated that NF-${\kappa}B$ protein may regulate expression of the Nanog gene. We used human embryonic carcinoma (EC) cells as a model system of ES cells and confirmed decrease of Nanog and increase of NF-${\kappa}B$ upon differentiation induced by retinoic acid. Although deletion analysis on the DNA fragment including NF-${\kappa}B$ binding site suggested involvement of NF-${\kappa}B$ in the negative regulation of the promoter, site-directed mutation of NF-${\kappa}B$ binding site had no effect on the Nanog promoter activity. Furthermore, no direct association of NF-${\kappa}B$ with the Nanog promoter was detected during differentiation. Therefore, we conclude that NF-${\kappa}B$ protein may not be involved in transcriptional regulation of Nanog gene expression in EC cells and possibly in ES cells.

• Korean Journal of Veterinary Research
• /
• v.54 no.3
• /
• pp.179-184
• /
• 2014

This study investigated the anatomy of the nutrient foramen (NF) in German Shepherds by recording the number, site, position, and direction of penetration of the nutrient canal (NC) in the humerus, radius, and ulna of 50 individuals. The site index of the nutrient foramen (SI) was calculated as the ratio of the length to the NF site from the proximal end to the greatest length of the bone. The NF diameter was measured using different sized needles. Most humeri had only one NF on the caudal surface, particularly on the lateral supracondylar crest, or distal cranial surface. All radii had one NF, usually on the caudal surface, while most ulnae had one NF located on either the cranial or lateral surfaces. The SI and NF diameters were 58.0~59.5% and 0.73~0.78 mm in the humerus, 30.4~30.9% and 0.74~0.76 mm in the radius, and 29.3~29.8% and 0.67~0.68 mm in the ulna, respectively. With the exception of the relatively proximal NF of the radius, the direction of penetration followed Berard's rule. This study provides novel information on the location and diameter of the NF and direction of the NC in the long bones of the pectoral limb of German Shepherds.

• Journal of the Semiconductor & Display Technology
• /
• v.17 no.4
• /
• pp.97-100
• /
• 2018

$NF_3$ Plasma etching of silicon was conducted by injecting only $NF_3$ gas into reactive ion etching. $NF_3$ Plasma etching was done in intermediate pressure. Silicon etching by $NF_3$ plasma in reactive ion etching was diagnosed through residual gas analyzer and optical emission spectroscopy. In plasma etching, optical emission spectroscopy is generally used to know what kinds of species in plasma. Also, residual gas analyzer is mainly to know the byproducts of etching process. Through experiments, the results of optical emission spectroscopy during silicon etching by $NF_3$ plasma was analyzed with connecting the results of etch rate of silicon and residual gas analyzer. It was confirmed that $NF_3$ plasma etching of silicon in reactive ion etching accords with the characteristic of reactive ion etching.

• Tuberculosis and Respiratory Diseases
• /
• v.51 no.4
• /
• pp.315-324
• /
• 2001

Background : IL-8 is a potent chemotactic cytokine that plays an important role in the host defense mechanism against M. tuberculosis by recruiting inflammatory cells to the site of the infection. Lung epithelial cells, as well as alveolar macrophages are known to produce IL-8 in response to M. tuberculosis. IL-8 gene expression is mainly regulated on the level of transcription by NF-${\kappa}B$. This study investigated whether or not A549 cells produce IL-8 in NF-${\kappa}B$ dependent mechanism in response to macrophages phagocytosing M. tuberculosis. Methods : Peripheral blood monocytes that were obtained from healthy donors were cultured for 24 h with M. tuberculosis and a conditioned medium(CoMTB) was obtained. As a negative control, the conditioned medium without M. tuberculosis (CoMCont) was used. A549 cells were stimulated with M. tuberculosis, CoMCont and CoMTB and the IL-8 concentration in the culture media was measured by ELISA. The CoMTB induced IL-8 mRNA expression in the A549 cells was evaluated using RT-PCR, and CoMTB induced $I{\kappa}B{\alpha}$ degradation was measured using western blot analysis. CoMTB induced nuclear translocation and DNA binding of NF-${\kappa}B$ was also examined using an electrophoretic mobility shift assay(EMSA), and the CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity was measured using a luciferase reporter gene assay. Results : CoMTB induced IL-8 production by A549 cells($46.8{\pm}4.8\;ng/ml$) was higher than with direct stimulation with M. tuberculosis ($6.8{\pm}2.9\;ng/ml$). CoMTB induced IL-8 mRNA expression increased after 2 h of stimulation and was sustained for 24 h. $I{\kappa}B{\alpha}$ was degraded after 10 min of CoMTB stimulation and reappeared by 60 min. CoMTB stimulated the nuclear translocation and DNA binding of NF-${\kappa}B$. The CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity($13.6{\pm}4.3$ times control) was higher than either CoMCont($2.0{\pm}0.6$ times control) or M. tuberculosis ($1.4{\pm}0.6$ times control). Conclusion : A conditioned medium of peripheral blood monocytes phagocytosing M. tuberculosis stimulates NF-${\kappa}B$ dependent IL-8 production by the lung epithelial cells.

• Membrane Journal
• /
• v.19 no.4
• /
• pp.317-323
• /
• 2009

In this study the nanofiltration (NF) membrane treatment of a nitric acid waste solutions containing $Pb^{+2}$ heavy metal ion discharging from the etching processes of an electronics and semiconductors industry has been studied for the purpose of recycling of nitric acid etching solutions. Three kinds of NF membranes (General Electric Co. Duraslick NF-4040 membrane, Dow Co. Filmtec LP-4040 membrane and Koch Co. SelRO MPS-34 4040 membrane) were tested for their separation efficiency (total rejection) of $Pb^{+2}$ ion and membrane stability in nitric acid solution. NF experiments were carried out with a dead-end membrane filtration laboratory system. The membrane permeate flux was increased with the increasing storage time in nitric acid solution and lowering pH of acid solution because of the enhancing of NF membrane damage by nitric acid. The membrane stability in nitric acid solution was more superior in the order of Filmtec LP-4040 < Duraslick NF-4040 < SelRO MPS-34 4040 membrane. The total rejection of Pb+2 ion was decreased with the increasing storage time in nitric acid solution and lowering the pH of acid solution. The total rejection of $Pb^{+2}$ ion after 4 months NF treatment was decreased from 95% initial value to 20% in the case of Duraslick NF-4040 membrane, from 85% initial value to 65% in the case of SelRO MPS-34 4040 membrane and from 90% initial value to 10% in the case of Filmtec LP-4040 membrane. These results showed that SelRO MPS-34 4040 NF membrane was more suitable for the treatment of an acidic etching waste solutions containing heavy metal ions.

• Tuberculosis and Respiratory Diseases
• /
• v.65 no.6
• /
• pp.476-486
• /
• 2008

Background: TRAIL (TNF-related apoptosis inducing ligand) is a newly identified member of the TNF gene family which appears to have tumor-selective cytotoxicity due to the distinct decoy receptor system. TRAIL has direct access to caspase machinery and induces apoptosis regardless of p53 phenotype. Therefore, TRAIL has a therapeutic potential in lung cancer which frequently harbors p53 mutation in more than 50% of cases. However, it was shown that TRAIL also could activates $NF-{\kappa}B$ in some cell lines which might inhibit TRAIL-induced apoptosis. This study was designed to investigate whether TRAIL can activate $NF-{\kappa}B$ in lung cancer cell lines relatively resistant to TRAIL-induced apoptosis and inhibition of $NF-{\kappa}B$ activation using proteasome inhibitor MG132 which blocks $I{\kappa}B{\alpha}$ degradation can sensitize lung cancer cells to TRAIL-induced apoptosis. Methods: A549 (wt p53) and NCI-H1299 (null p53) lung cancer cells were used and cell viability test was done by MTT assay. Apoptosis was confirmed with Annexin V assay followed by FACS analysis. To study $NF-{\kappa}B$-dependent transcriptional activation, a luciferase reporter gene assay was used after making A549 and NCI-H1299 cells stably transfected with IgG ${\kappa}-NF-{\kappa}B$ luciferase construct. To investigate DNA binding of $NF-{\kappa}B$ activated by TRAIL, electromobility shift assay was used and supershift assay was done using anti-p65 antibody. Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation. Results: A549 and NCI-H1299 cells were relatively resistant to TRAIL-induced apoptosis showing only 20~30% cell death even at the concentration 100 ng/ml, but MG132 ($3{\mu}M$) pre-treatment 1 hour prior to TRAIL addition greatly increased cell death more than 80%. Luciferase assay showed TRAIL-induced $NF-{\kappa}B$ transcriptional activity in both cell lines. Electromobility shift assay demonstrated DNA binding complex of $NF-{\kappa}B$ activated by TRAIL and supershift with p65 antibody. $I{\kappa}B{\alpha}$ degradation was proven by western blot. MG132 completely blocked both TRAIL-induced $NF-{\kappa}B$ dependent luciferase activity and DNA binding of $NF-{\kappa}B$. Conclusion: This results suggest that inhibition of $NF-{\kappa}B$ can be a potentially useful strategy to enhance TRAIL-induced tumor cell killing in lung cancer.

• Journal of Bio-Environment Control
• /
• v.31 no.4
• /
• pp.460-467
• /
• 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.

• The Journal of Pediatrics of Korean Medicine
• /
• v.25 no.1
• /
• pp.28-39
• /
• 2011

Objectives: SaengRyoSaMulTang is a herbal formula in Oriental Medicine, known anti-allergens. However, its mechanism and cellular targets have not been found yet. Thus the study has developed to investigate the suppressive effect of SaengRyoSaMulTang. Methods: In the study, cellular viability, IL-4, IL-13 mRNA expression, IL-4, IL-13 production, manifestations of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}$B p65 transcription factors were examined by Real-Time PCR, ELISA analysis and western blotting. Results: As a result of treating with SaengRyoSaMulTang extract(SRSMT), the study has shown that the amount of Th2 cytokines, which include PI induced IL-4 and IL-13, plays a significant role in suppressing effect. RBL-2H3 mast cells significantly suppressed the PI-induced Th2 cytokine production including IL-4 and IL-13 in a dose dependent manner. PI-induced IL-4 and IL-13 production was significantly suppressed by SRSMT intervention. Western blot analysis of transcription factors involving IL-4 and IL-13 expression also revealed a prominent decreases of mast cell's specific transcription factors including GATA-1, GATA-2, NF-AT2, c-Jun and c-Fos, but NF-${\kappa}$B p65. Conclusions: The study suggests that the anti-allergenic activities of SRSMT may regulate the transcription factors GATA-1, GATA-2, NF-AT2, c-Jun and c-Fos inhibiting Th2 cytokines IL-4 and IL-13 in mast cells.

• BMB Reports
• /
• v.44 no.4
• /
• pp.267-272
• /
• 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.

• The Korean Journal of Physiology and Pharmacology
• /
• v.25 no.4
• /
• pp.307-319
• /
• 2021

Dysregulation of the Wnt pathway causes various diseases including cancer, Parkinson's disease, Alzheimer's disease, schizophrenia, osteoporosis, obesity and chronic kidney diseases. The modulation of dysregulated Wnt pathway is absolutely necessary. In the present study, we evaluated the anti-inflammatory effect and the mechanism of action of Wnt-C59, a Wnt signaling inhibitor, in lipopolysaccharide (LPS)-stimulated epithelial cells and macrophage cells. Wnt-C59 showed a dose-dependent anti-inflammatory effect by suppressing the expression of proinflammatory cytokines including IL6, CCL2, IL1A, IL1B, and TNF in LPS-stimulated cells. The dysregulation of the Wnt/β-catenin pathway in LPS stimulated cells was suppressed by WntC59 treatment. The level of β-catenin, the executor protein of Wnt/β-catenin pathway, was elevated by LPS and suppressed by Wnt-C59. Overexpression of β-catenin rescued the suppressive effect of Wnt-C59 on proinflammatory cytokine expression and nuclear factor-kappa B (NF-κB) activity. We found that the interaction between β-catenin and NF-κB, measured by co-immunoprecipitation assay, was elevated by LPS and suppressed by Wnt-C59 treatment. Both NF-κB activity for its target DNA binding and the reporter activity of NF-κB-responsive promoter showed identical patterns with the interaction between β-catenin and NF-κB. Altogether, our findings suggest that the anti-inflammatory effect of Wnt-C59 is mediated by the reduction of the cellular level of β-catenin and the interaction between β-catenin and NF-κB, which results in the suppressions of the NF-κB activity and proinflammatory cytokine expression.