• Title/Summary/Keyword: Line Marker

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Relationship between Cancer Stem Cell Marker CD133 and Cancer Germline Antigen Genes in NCI-H292 Lung Cancer Cells

  • Ko, Taek Yong;Kim, Jong In;Lee, Sang Ho
    • Journal of Chest Surgery
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    • v.53 no.1
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    • pp.22-27
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    • 2020
  • Background: Previous studies have shown that lung cancer stem cells express CD133 and that certain cancer stem cells express cancer germline antigens (CGAs). The transcriptional regulation of CD133 is complicated and poorly understood. We investigated CD133 and CGA expression in a non-small cell lung cancer cell line. Methods: The expression levels of CD133 and CGAs (MAGE-6, GAGE, SSX, and TRAG-3) were measured in an NCI-H292 lung cancer cell line. The methylation status of the CD133 gene promoter region was analyzed. The expression levels and promoter methylation statuses of CD133 and CGAs were confirmed by treatment with the demethylating agent 5-aza-2'-deoxycytidine (ADC). Results: After treatment with ADC, CD133 expression was no longer detected. MAGE-6 and TRAG-3 were detected before ADC treatment, while GAGE and SSX were not detected. ADC treatment upregulated MAGE-6 and TRAG-3 expression, while GAGE expression was still undetected after treatment, and only weak SSX expression was observed. GAGE expression was not correlated with expression of CD133, while the levels of expression of MAGE-6, TRAG-3, and SSX were inversely correlated with CD133 expression. Conclusion: These results showed that CD133 expression can be regulated by methylation. Thus, the demethylation of the CD133 promoter may compromise the treatment of lung cancer by inactivating cancer stem cells and/or activating CGAs.

Effect of implant surface microtopography by hydroxyapatite grit-blasting on adhesion, proliferation, and differentiation of osteoblast-like cell line, MG-63

  • Park, Sung-Jae;Bae, Sang-Bum;Kim, Su-Kyoung;Eom, Tae-Gwan;Song, Seung-Il
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.3
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    • pp.214-224
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    • 2011
  • Objective: This study examined the potential of the in vitro osteogenesis of microtopographically modified surfaces, RBM (resorbable blasting media) surfaces, which generate hydroxyapatite grit-blasting. Methods: RBM surfaces were modified hydroxyapatite grit-blasting to produce microtopographically modified surfaces and the surface morphology, roughness or elements were examined. To investigate the potential of the in vitro osteogenesis, the osteoblastic cell adhesion, proliferation, and differentiation were examined using the human osteoblast-like cell line, MG-63 cells. Osteoblastic cell proliferation was examined as a function of time. In addition, osteoblastic cell differentiation was verified using four different methods of an ALP activity assay, a mineralization assay using alizarin red-s staining, and gene expression of osteoblastic differentiation marker using RT-PCR or ELISA. Results: Osteoblastic cell adhesion, proliferation and ALP activity was elevated on the RBM surfaces compared to the machined group. The cells exhibited a high level of gene expression of the osteoblastic differentiation makers (osteonectin, type I collagen, Runx-2, osterix). imilar data was represented in the ELISA produced similar results in that the RBM surface increased the level of osteocalcin, osteopontin, TGF-beta1 and PGE2 secretion, which was known to stimulate the osteogenesis. Moreover, alizarin red-s staining revealed significantly more mineralized nodules on the RBM surfaces than the machined discs. Conclusion: RBM surfaces modified with hydroxyapatite grit-blasting stimulate the in vitro osteogenesis of MG-63 cells and may accelerate bone formation and increase bone-implant contact.

Ginsenoside Rh2 epigenetically regulates cell-mediated immune pathway to inhibit proliferation of MCF-7 breast cancer cells

  • Lee, Hyunkyung;Lee, Seungyeon;Jeong, Dawoon;Kim, Sun Jung
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.455-462
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    • 2018
  • Background: Ginsenoside Rh2 has been known to enhance the activity of immune cells, as well as to inhibit the growth of tumor cells. Although the repertoire of genes regulated by Rh2 is well-known in many cancer cells, the epigenetic regulation has yet to be determined, especially for comprehensive approaches to detect methylation changes. Methods: The effect of Rh2 on genome-wide DNA methylation changes in breast cancer cells was examined by treating cultured MCF-7 with Rh2. Pyrosequencing analysis was carried out to measure the methylation level of a global methylation marker, LINE1. Genome-wide methylation analysis was carried out to identify epigenetically regulated genes and to elucidate the most prominent signaling pathway affected by Rh2. Apoptosis and proliferation were monitored to examine the cellular effect of Rh2. Results: LINE1 showed induction of hypomethylation at specific CpGs by 1.6-9.1% (p < 0.05). Genome-wide methylation analysis identified the "cell-mediated immune response"-related pathway as the top network. Cell proliferation of MCF-7 was retarded by Rh2 in a dose-dependent manner. Hypermethylated genes such as CASP1, INSL5, and OR52A1 showed downregulation in the Rh2-treated MCF-7, while hypomethylated genes such as CLINT1, ST3GAL4, and C1orf198 showed upregulation. Notably, a higher survival rate was associated with lower expression of INSL5 and OR52A1 in breast cancer patients, while with higher expression of CLINT1. Conclusion: The results indicate that Rh2 induces epigenetic methylation changes in genes involved in immune response and tumorigenesis, thereby contributing to enhanced immunogenicity and inhibiting the growth of cancer cells.

Vanishing Line based Lane Detection for Augmented Reality-aided Driver Induction

  • Yun, Jeong-Rok;Lee, Dong-Kil;Chun, Sung-Kuk;Hong, Sung-Hoon
    • Journal of the Korea Society of Computer and Information
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    • v.24 no.1
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    • pp.73-83
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    • 2019
  • In this paper, we propose the augmented reality(AR) based driving navigation based on robust lane detection method to dynamic environment changes. The proposed technique uses the detected lane position as a marker which is a key element for enhancing driving information. We propose Symmetrical Local Threshold(SLT) algorithm which is able to robustly detect lane to dynamic illumination environment change such as shadows. In addition, by using Morphology operation and Connected Component Analysis(CCA) algorithm, it is possible to minimize noises in the image, and Region Of Interest(ROI) is defined through region division using a straight line passing through several vanishing points We also propose the augmented reality aided visualization method for Interchange(IC) and driving navigation using reference point detection based on the detected lane coordinates inside and outside the ROI. Validation experiments were carried out to assess the accuracy and robustness of the proposed system in vairous environment changes. The average accuracy of the proposed system in daytime, nighttime, rainy day, and cloudy day is 79.3% on 4600 images. The results of the proposed system for AR based IC and driving navigation were also presented. We are hopeful that the proposed research will open a new discussion on AR based driving navigation platforms, and thus, that such efforts will enrich the autonomous vehicle services in the near future.

Anti-allergic Effect of the Fermented Angelicae Gigantis Radix in Human Mast Cell Line HMC-1 (발효처리한 당귀의 항알레르기 효능에 대한 연구)

  • Seo, Min-Jun;Park, Jin-Han;Lee, Je-Hyun
    • The Korea Journal of Herbology
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    • v.28 no.5
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    • pp.39-44
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    • 2013
  • Objectives : Allergy is an immune dysfunction caused by degranulation from mast cells in the early phase of allergic disease. The purpose of this study was to investigate the anti-allergic effect of fermented Angelicae gigantis Radix in human mast cell line, HMC-1. Method : The Angelicae gigantis Radix was fermented by Lactobacillus acidophilus. The cell toxicity of fermented Angelicae gigantis Radix(FAGR) was determined by MTT assay. The release of ${\beta}$-hexosaminidase from HMC-1 stimulated by phorbol-12-myristate 13-acetate (PMA) plus A23187 was determined by ${\beta}$-hexosaminidase assay. Also, the concentrations of cytokines (interleukin-$1{\beta}$, -6, -8 and tumor necrosis factor-alpha) were measured by enzyme-linked immunosorbent assay. The gene expression of COX-2 from HMC-1 stimulated by phorbol-12-myristate 13-acetate (PMA) plus A23187 was determined by reverse transcription polymerase chain reaction. The release of histamine on substance P-stimulated HMC-1 was measured by histamine assay. Result : The FAGR suppressed the release of ${\beta}$-hexosaminidase, a marker of degranulation, from HMC-1 stimulated by PMA plus A23187. The FAGR inhibited the production of interleukin-$1{\beta}$, -6, -8 and tumor necrosis factor-alpha. The FAGR inhibited the expression of COX-2 mRNA. The FAGR suppressed the release of histamine on substance P-stimulated HMC-1. Conclusion : These results provide that FAGR may be beneficial in the treatment of allergic inflammatory disease.

Post HCV Infection Due to MX Gene Stimulation Produced Post Treatment with Imported and Locally Produced Egyptian Biosimilar IFN

  • Mohamed, Shereen H;Mahmoud, Nora F;Mohamed, Aly F;Kotb, Nahla S
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5635-5641
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    • 2015
  • Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.

Expression of Murine GM-CSF in Recombinant Aspergillus niger

  • Kim, Nyoung-Ji;Kwon, Tae-Ho;Jang, Yong-Suk;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.287-292
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    • 2000
  • Recombinant Aspergillus niger was constructed to express and secrete a biologically active murine granulaocyte macrophage-colony stimulating factor (mGM-CSF). A 500 bp fragment encoding the signal peptide and terminator of glyceraldehyde-3-phosphate dehydrogenase (gpd). The hygromycin phosphotrasferase gene (hph) was used as a selection marker for the fungal transformants. An expression vector was introduced into A. niger ATCC 9642, and a Northern blot analysis indicated the presence of a considerable amount of transcripts from the introduced mGM-CSF. The biological activity of recombinant mGM-CSF (rmGM-CSF) isolated from the culture filtrate was confirmend by measuring the proliferationof the GM-CSF dependent FDC-P1 cell line. It appeared that rmGM-CSF was amenable to the proteolytic activity produced by A. niger, since biological actibity was only observed when the transformants were grown in a protease-repressing medium, and the activity of rmGM-CSF dramatically decreased with an increase of age of the culture. The yield of rmGM-CSF, as determined by ELISA. was 640 ng/l of culture filtrate. Accordingly, its specific activity is estimated to be approximately two-and-a-half times higher than that of a commercial preparation from E. coli.

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Identification of Novel Clubroot Resistance Loci in Brassic rapa

  • Pang, Wenxing;Chen, Jingjing;Yu, Sha;Shen, Xiangqun;Zhang, Chunyu;Piao, Zhongyun
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.42-42
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    • 2015
  • Plasmodiophora brassicae, the causal agent of clubroot disease, does the most serious damage to the Brassica crops. The limited control approaches make that the identification of clubroot resistance (CR) is more important for developing CR cultivars of the Brassica crops. So far, 8 CR loci were mapped. However, the variation of P. brassicae leads to the rapid erosion of its resistance. To identify novel CR genes, we employed three mapping population, derived from crosses between Chinese cabbage and turnip inbred lines ($59-1{\times}ECD04$ and $BJN3-1{\times}Siloga$) or between Chinese cabbage inbred lines ($BJN3-1{\times}85-I-II$), to perform QTL analysis. Totally, 8 CR loci were indentified and showed race-specific resistance. Physical mapping of these 8 loci suggested that 4 were located previously mapped position, indicating they might be the same allele or different alleles of the same genes. Other 4 loci were found to be novel. Further, CR near isogenic line carrying each CR locus was developed based on the marker assisted selection. Verification of these CR loci was underway. Identification of these novel CR genes would facilitate to breed broad-spectrum and durable CR cultivars of B. rapa by pyramiding strategies.

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Digital Content Interactions Using RFID/NFC-based Tangible Interfaces in Augmented Reality Environments (증강현실 환경하에서 RFID/NFC 기반의 탠저블 인터페이스를 활용한 디지털 콘텐츠 상호작용)

  • Seo, Dong Woo;Lee, Jae Yeol;Kim, Jae Sung
    • Korean Journal of Computational Design and Engineering
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    • v.20 no.2
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    • pp.159-170
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    • 2015
  • Radio-Frequency Identification (RFID) or Near Field Communication (NFC) technology has many advantages over other visual interfaces since it does not require line-of-sight alignment, can identify multiple tags simultaneously, and does not destroy the integrity of original objects. In addition, smart devices such as smartphone and smartpad have NFC/RFID readers which can provide mobile and natural interactions with digital and physical contents. Augmented reality has an excellent visual interaction capability with digital contents in a real environment by embedding digital contents into the physical world. In this paper, we propose a new approach to digital content interactions using RFID/NFC-based tangible interfaces in augmented reality environments that utilize invisible interfaces in addition to marker-based visual interfaces. By combining the advantages of invisible and visual interfaces, more intuitive interactions with digital contents can be provided, which can remove the difficulty of using typical AR paddles that are widely used in AR interactions. In particular, a semantic AR ontology is defined to provide more convenient interactions. Through the semantic ontology-based inferencing, physical querying and filtering are effectively supported. We will show the effectiveness and advantage of the proposed approach by demonstrating implementation results.

Genetic Linkage Mapping of RAPD Markers Segregating in Korean Ogol Chicken - White Leghorn Backcross Population

  • Hwang, K.C.;Song, K.D.;Kim, T.H.;Jeong, D.K.;Sohn, S.H.;Lillehoj, H.S.;Han, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.3
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    • pp.302-306
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    • 2001
  • This study was carried out to construct mapping population and to evaluate the methods involved, including polymorphic DNA marker system and appropriate statistical analysis. As an initial step to establish chicken genome mapping project, White Leghorn (WL) and Korean Ogol chicken (KOC) were used for generating backcross population. From 8 initial parents, total 280 backcross progenies were obtained and 40 were used for genotyping and linkage analysis. For development of novel polymorphic markers for KOC, Random Amplified Polymorphic DNA (RAPD) markers specific for this chicken line were generated. Also included in this study were six microsatellite markers from East Lansing map as reference loci. For segregation analysis, 15 RAPD markers and 6 microsatellites were used to genotype the backcross population. Among the RAPD markers that we developed, 2 pairs of markers were identified to be linked and another 4 RAPD markers showed linkage with microsatellites of known map. In summary, this study showed that our backcross population generated from the mating of KOC to WL serves as a valuable genetic resource for genotyping. Furthermore, RAPD markers are proved to be valuable in linkage mapping analysis.