• Title/Summary/Keyword: Multiple-Chain

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Construction and Production of Concatameric Human TNF Receptor-Immunoglobulin Fusion Proteins

  • Yim, Su-Bin;Chung, Yong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.81-89
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    • 2004
  • Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and lymphotoxin-$\alpha$ (LT-$\alpha$, TNF-$\beta$) can initiate and perpetuate human diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA), and insulin-dependent diabetes mellitus (IDDM). TNFs can be blocked by the use of soluble TNF receptors. However, since monomeric soluble receptors generally exhibit low affinity or function as agonists, the use of monomeric soluble receptors has been limited in the case of cytokines such as TNF-$\alpha$, TNF-$\alpha$, interleukin (IL)-1, IL-4, IL-6, and IL-13, which have adapted to a multi component receptor system. For these reasons, very high-affinity inhibitors were created for the purpose of a TNFs antagonist to bind the TNFR and trigger cellular signal by using the multistep polymerase chain reaction method. First, recombinant simple TNFR-Ig fusion proteins were constructed from the cDNA sequences encoding the extracellular domain of the human p55 TNFR (CD120a) and the human p75 TNFR (CD120b), which were linked to hinge and constant regions of human $IgG_1$ heavy chain, respectively using complementary primers (CP) encoding the complementary sequences. Then, concatameric TNFR-Ig fusion proteins were constructed using recombinant PCR and a complementary primer base of recombinant simple TNFR-Ig fusion proteins. For high level expression of recombinant fusion proteins, Chinese hamster ovary (CHO) cells were used with a retroviral expression system. The transfected cells produced the simple concatameric TNFR-Ig fusion proteins capable of binding TNF and inactivating it. These soluble versions of simple concantameric TNFR-Ig fusion proteins gave rise to multiple forms such as simple dimers and concatameric homodimers. Simple TNFR-1g fusion proteins were shown to have much more reduced TNF inhibitory activity than concatameric TNFR-Ig fusion proteins. Concatameric TNFR-Ig fusion proteins showed higher affinity than simple TNFR-Ig fusion proteins in a receptor inhibitor binding assay (RIBA). Additionally, concatameric TNFR-Ig fusion proteins were shown to have a progressive effect as a TNF inhibitor compared to the simple TNFR-Ig fusion proteins and conventional TNFR-Fc in cytotoxicity assays, and showed the same results for collagen induced arthritis (CIA) in mice in vivo.

A Study on Regionalization of Parameters for Sacramento Continuous Rainfall-Runoff Model Using Watershed Characteristics (유역특성인자를 활용한 Sacramento 장기유출모형의 매개변수 지역화 기법 연구)

  • Kim, Tae-Jeong;Jeong, Ga-In;Kim, Ki-Young;Kwon, Hyun-Han
    • Journal of Korea Water Resources Association
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    • v.48 no.10
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    • pp.793-806
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    • 2015
  • The simulation of natural streamflow at ungauged basins is one of the fundamental challenges in hydrology community. The key to runoff simulation in ungauged basins is generally involved with a reliable parameter estimation in a rainfall-runoff model. However, the parameter estimation of the rainfall-runoff model is a complex issue due to an insufficient hydrologic data. This study aims to regionalize the parameters of a continuous rainfall-runoff model in conjunction with a Bayesian statistical technique to consider uncertainty more precisely associated with the parameters. First, this study employed Bayesian Markov Chain Monte Carlo scheme for the estimation of the Sacramento rainfall-runoff model. The Sacramento model is calibrated against observed daily runoff data, and finally, the posterior density function of the parameters is derived. Second, we applied a multiple linear regression model to the set of the parameters with watershed characteristics, to obtain a functional relationship between pairs of variables. The proposed model was also validated with gauged watersheds in accordance with the efficiency criteria such as the Nash-Sutcliffe efficiency, index of agreement and the coefficient of correlation.

The Effects of Environmental Dynamism on Supply Chain Commitment in the High-tech Industry: The Roles of Flexibility and Dependence (첨단산업의 환경동태성이 공급체인의 결속에 미치는 영향: 유연성과 의존성의 역할)

  • Kim, Sang-Deok;Ji, Seong-Goo
    • Journal of Global Scholars of Marketing Science
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    • v.17 no.2
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    • pp.31-54
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    • 2007
  • The exchange between buyers and sellers in the industrial market is changing from short-term to long-term relationships. Long-term relationships are governed mainly by formal contracts or informal agreements, but many scholars are now asserting that controlling relationship by using formal contracts under environmental dynamism is inappropriate. In this case, partners will depend on each other's flexibility or interdependence. The former, flexibility, provides a general frame of reference, order, and standards against which to guide and assess appropriate behavior in dynamic and ambiguous situations, thus motivating the value-oriented performance goals shared between partners. It is based on social sacrifices, which can potentially minimize any opportunistic behaviors. The later, interdependence, means that each firm possesses a high level of dependence in an dynamic channel relationship. When interdependence is high in magnitude and symmetric, each firm enjoys a high level of power and the bonds between the firms should be reasonably strong. Strong shared power is likely to promote commitment because of the common interests, attention, and support found in such channel relationships. This study deals with environmental dynamism in high-tech industry. Firms in the high-tech industry regard it as a key success factor to successfully cope with environmental changes. However, due to the lack of studies dealing with environmental dynamism and supply chain commitment in the high-tech industry, it is very difficult to find effective strategies to cope with them. This paper presents the results of an empirical study on the relationship between environmental dynamism and supply chain commitment in the high-tech industry. We examined the effects of consumer, competitor, and technological dynamism on supply chain commitment. Additionally, we examined the moderating effects of flexibility and dependence of supply chains. This study was confined to the type of high-tech industry which has the characteristics of rapid technology change and short product lifecycle. Flexibility among the firms of this industry, having the characteristic of hard and fast growth, is more important here than among any other industry. Thus, a variety of environmental dynamism can affect a supply chain relationship. The industries targeted industries were electronic parts, metal product, computer, electric machine, automobile, and medical precision manufacturing industries. Data was collected as follows. During the survey, the researchers managed to obtain the list of parts suppliers of 2 companies, N and L, with an international competitiveness in the mobile phone manufacturing industry; and of the suppliers in a business relationship with S company, a semiconductor manufacturing company. They were asked to respond to the survey via telephone and e-mail. During the two month period of February-April 2006, we were able to collect data from 44 companies. The respondents were restricted to direct dealing authorities and subcontractor company (the supplier) staff with at least three months of dealing experience with a manufacture (an industrial material buyer). The measurement validation procedures included scale reliability; discriminant and convergent validity were used to validate measures. Also, the reliability measurements traditionally employed, such as the Cronbach's alpha, were used. All the reliabilities were greater than.70. A series of exploratory factor analyses was conducted. We conducted confirmatory factor analyses to assess the validity of our measurements. A series of chi-square difference tests were conducted so that the discriminant validity could be ensured. For each pair, we estimated two models-an unconstrained model and a constrained model-and compared the two model fits. All these tests supported discriminant validity. Also, all items loaded significantly on their respective constructs, providing support for convergent validity. We then examined composite reliability and average variance extracted (AVE). The composite reliability of each construct was greater than.70. The AVE of each construct was greater than.50. According to the multiple regression analysis, customer dynamism had a negative effect and competitor dynamism had a positive effect on a supplier's commitment. In addition, flexibility and dependence had significant moderating effects on customer and competitor dynamism. On the other hand, all hypotheses about technological dynamism had no significant effects on commitment. In other words, technological dynamism had no direct effect on supplier's commitment and was not moderated by the flexibility and dependence of the supply chain. This study makes its contribution in the point of view that this is a rare study on environmental dynamism and supply chain commitment in the field of high-tech industry. Especially, this study verified the effects of three sectors of environmental dynamism on supplier's commitment. Also, it empirically tested how the effects were moderated by flexibility and dependence. The results showed that flexibility and interdependence had a role to strengthen supplier's commitment under environmental dynamism in high-tech industry. Thus relationship managers in high-tech industry should make supply chain relationship flexible and interdependent. The limitations of the study are as follows; First, about the research setting, the study was conducted with high-tech industry, in which the direction of the change in the power balance of supply chain dyads is usually determined by manufacturers. So we have a difficulty with generalization. We need to control the power structure between partners in a future study. Secondly, about flexibility, we treated it throughout the paper as positive, but it can also be negative, i.e. violating an agreement or moving, but in the wrong direction, etc. Therefore we need to investigate the multi-dimensionality of flexibility in future research.

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Locational Characteristics of Knowledge Service Industry and Related Employment Opportunity Estimation in the Seoul Metropolitan Area (서울대도시권 지식서비스산업의 입지적 특성과 관련 업종별 고용기회 예측)

  • Park, So Hyun;Lee, Keumsook
    • Journal of the Economic Geographical Society of Korea
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    • v.19 no.4
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    • pp.694-711
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    • 2016
  • This study analyzes the spatial characteristics of knowledge industry which has shown relatively rapid growth in the low-growth economy situation in recent years. In particular, we catch hold of the locational characteristics of the knowledge service industry which occupies the highest ratio by professional-expert jobs favoured by young generations, as well as estimate their occupational employment opportunities. By applying Location Quotient(LQ) and LISA, we reveal the spatial distribution patterns of publishing business, information service business and education service business in the Seoul Metropolitan area, and examine the changes in the spatial patterns during the last ten years. In order to understand the socio-economic factors which explain their locations, we apply the stepwise multiple regression analysis. Furthermore, we predict the changes distribution of Knowledge service industrial employment by applying Markov Chain Model. As the result, we found their clusters at the specific locations, while there is the significant variations in the socio-economic variables related their locations respectively. The related job opportunities of the knowledge service businesses in the Seoul Metropolitan area are predicted steady growth trend for the next four years, even though dull or stagnant trend is expected for other industries. This study provides basic resources to the planning for young generation employment problem.

Establishment of 8-Azaguanine Resistant Human Plasmacytoma Cell Line (8-Azaguanine 내성 인형 형질세포종 세포주의 확립)

  • Cha, Chang-Yong;Hwang, Eung-Soo;Kook, Yoon-Ho;Lim, Dong-Kyun;Cho, Han-Ik;Park, Myung-Hee;Kim, Noe-Kyung;Chang, Woo-Hyun;Lee, Mun-Ho
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.3
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    • pp.399-406
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    • 1986
  • This study was performed to establish human plasmacytoma cell line as the partner cells for producing human hybridoma. Bone marrow cells from a multiple myeloma patient from Seoul National University Hospital, Korea were cultured and established as the cell line, named as HMC-BM4. HMC-BM4 cells were cultivated in RPMI 1640 media containing 8-azaguanine(8-AG; gradually increasing concentration from $1\;{\mu}g/ml$ to $20\;{\mu}g/ml$). 8-AG resistant cells were collected and cloned by limiting dilution. Each clone was divided and tested to die in hypoxanthine, aminopterine and thymidine (HAT) selection media. Finally one clone was selected and named as HMC-AR, which was sensitive to HAT selection media. HMC-AR cells showed typical morphology of plamacytoma in Wright staining. No cell formed the rosette with sheep erythrocytes. Surface membrane $\mu$ heavy chain was detected in 20% of HMC-AR cells and cytoplasmic $\mu$ heavy chain in 90% of them by direct immunofluorescent staining. Ia-like antigen was found in 90% of HMC-AR cells by indirect immunofluorescent staining using anti-Ia-like antigen monoclonal antibody, 1BD9-2. And about $1.0\;{\mu}g/ml$ of human $\mu$ heavy chain was detected in the 3-day culture supernatant of HMC-AR cells. 88% of cells contained 46 chromosomes. Mycoplasma was not detected in HMC-AR cells by Hoechst 33258 staining. This cell line would be used for making hybridomas secreting human monoclonal antibody.

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Detection of Borrelia burgdorferi and Ehrlichiosis Agent in Ticks Collected in Korea Using Polymerase Chain Reaction (국내에서 채집한 진드기에서 중합효소연쇄반응을 이용한 라임병균 및 Ehrlichiosis 원인체의 검출)

  • 김종배;송혜원;박성언;박상욱;안준환;엄용빈;김영미
    • Biomedical Science Letters
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    • v.4 no.2
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    • pp.113-120
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    • 1998
  • To investigate the distribution of Borrelia burgdorferi and human granulocytic ehrlichiosis (HGE) agent in ticks, adult ixodid ticks of Ixodes spp. and Haemaphysalis spp. were collected from the high mountain areas of Kangwon Province. Using DNAs extracted and purified in the collected ticks, polymerase chain reaction (PCR) was performed to amplify the specific nucleotide sequences of both agents. Of the 516 ticks, a total of 68 (13.2%) ticks was positive for Borrelia burgdorferi sensu lato with PCR analysis (2 for B. burgdorferi sensu stricto; 1 for B. afzelii;33 for B. garinii; 8 for B. tanukii;4 for B. turdae). However a little more than half of PCR-positive ticks (37/68) was found to be positive in the southern blot analysis with Bl6S oligonucleotide probe. One hundred and one (19.2%) ticks were positive for Ehrlichia spp. in PCR, and a quarter of them (25/101) was positive in southern blot with El6S oligonucleotide probe. But none of them was found to be the DNA of HGE agent. And 0.6% (3/516) ticks were positive for both of B. burgdorferi sensu late and Ehrlirhia spp. These findings might implicate the possibility of the outbreak of Iyme borreliosis and ehrlichiosis in Korea, and more extensive studies may be need for the diagnosis of multiple tick-borne diseases.

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Melatonin inhibits the Migration of Colon Cancer RKO cells by Down-regulating Myosin Light Chain Kinase Expression through Cross-talk with p38 MAPK

  • Zou, Duo-Bing;Wei, Xiao;Hu, Ruo-Lei;Yang, Xiao-Ping;Zuo, Li;Zhang, Su-Mei;Zhu, Hua-Qing;Zhou, Qing;Gui, Shu-Yu;Wang, Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5835-5842
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    • 2015
  • Background: Melatonin, which is mainly produced by the pineal gland, has a good inhibitory effect on cell growth of multiple cancer types. However, the underlying molecular mechanisms of anti-tumor activity for colon cancer have not been fully elucidated. In this study, we investigated the effects of melatonin on migration in human colon cancer RKO cells and the potential molecular mechanisms. Materials and Methods: The viability of RKO cells was investigated by MTT assay after treatment with melatonin, SB203580 (p38 inhibitor) and phorbol 12-myristate 13-acetate (PMA, MAPK activator) alone or in combination for 48h. The effects of melatonin, and ML-7, a selective inhibitor of myosin light chain kinase (MLCK), and SB203580, and PMA on the migration of RKO cells were analyzed by in vitro scratch-wound assay. The relative mRNA levels of MLCK was assessed by real-time quantitative RT-PCR. Western blotting analysis was performed to examine the expression of MLCK, phosphorylation of myosin light chain (pMLC) and p38 (pp38). Results: The proliferation and migration of human colon cancer RKO cells were inhibited significantly after treatment with melatonin. The expression levels of MLCK and phosphorylation of MLC of RKO cells were reduced, and real-time quantitative RT-PCR showed that melatonin had significant effects on suppressing the expression of MLCK. Furthermore, the phosphorylation level of p38, which showed the same trend, was also reduced when cells were treated by melatonin. In addition, ML-7 (25umol/l) could down-regulate the phosphorylation of p38. Conclusions: Melatonin could inhibit the proliferation and migration of RKO cells, and further experiments confirmed that p38 MAPK plays an important role in regulating melatonin-induced migration inhibition through down-regulating the expression and activity of MLCK.

Inferring transmission routes of avian influenza during the H5N8 outbreak of South Korea in 2014 using epidemiological and genetic data (역학과 유전학적 데이터를 이용한 한국에서 2014년 발생한 H5N8 조류독감 전염경로의 유추)

  • Choi, Sang Chul
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.254-265
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    • 2018
  • Avian influenza recently damaged the poultry industry, which suffered a huge economic loss reaching billions of U.S. dollars in South Korea. Transmission routes of the pathogens would help plan to control and limit the spread of the devastating biological tragedy. Phylogenetic analyses of pathogen's DNA sequences could sketch transmission trees relating hosts with directed edges. The last decade has seen the methodological development of inferring transmission trees using epidemiological as well as genetic data. Here, I reanalyzed the DNA sequence data that had originated in the highly pathogenic avian influenza H5N8 outbreak of South Korea in 2014. The H5N8 viruses spread geographically contiguously from the origin of the outbreak, Jeonbuk. The Jeonbuk origin viruses were known to spread to four provinces neighboring Jeonbuk. I estimated the transmission tree of the host domestic and migratory wild birds after combining multiple runs of Markov chain Monte Carlo using a Bayesian method for inferring transmission trees. The estimated transmission tree, albeit with a rather large uncertainty in the directed edges, showed that the viruses spread from Jeonbuk through Chungnam to Gyeonggi. Domestic birds of breeder or broiler ducks were estimated to appear to be at the terminal nodes of the transmission tree. This observation confirmed that migratory wild birds played an important role as one of the main infection mediators in the avian influenza H5N8 outbreak of South Korea in 2014.

Prevalence of Human Papillomavirus Types and Phylogenetic Analysis of HPV-16 L1 Variants from Southern India

  • Kabekkodu, Shama Prasada;Bhat, Samatha;Pandey, Deeksha;Varghese, Vinay Koshy;Shukla, Vaibhav;Ghosh, Supriti;Kushtagi, Pralhad;Bhat, Parvati;Gopinath, Puthiya Mundayat;Satyamoorthy, Kapaettu
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.2073-2080
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    • 2015
  • Background: The human papillomavirus (HPV) and its variants show wide geographical distribution and have been reported to cause cervical lesions. With cervical neoplasia as the leading cancer in Indian women, the aim of the present study was to evaluate the multiple infection HPV type distribution and variant genotypes in cervical samples from the coastal Karnataka region, India. Materials and Methods: A total of 212 samples were screened by nested polymerase chain reaction using PGMY9/11 and GP5+/6+ primers. HPV positive samples were sequenced to identify the types and a phylogenetic tree was constructed using the neighbor-joining method. Results: Sequence analysis identified a total of 14 HPV types distributed in 20%, 73.3% and 82.5% of non-malignant, pre-malignant [low grade squamous intraepithelial lesion (LSIL) and high grade squamous intraepithelial lesion (HSIL)] and cervical cancer samples. The distribution of high risk HPV in cancer samples was HPV 16, 76.4%, HPV18, 11.7%, HPV81, 2.9%, HPV31, 1.4%, HPV35, 1.4% and HPV 45, 1.4%. Multiple infections were observed in 11.8% of tumor samples with HPV 16 contributing to 62.5% of cases. In non-malignant samples, 20% of HPV positive samples were detected with HPV16, 82.3%, HPV33, 5.8% and HPV58, 5.8% and very low incidence of multiple infections. Comparative phylogenetic analysis of HPV variants identified 9 HPV sequences as new papillomavirus species, predominantly classified as European lineage type. Conclusions: The findings for HPV infections associated with progression of cervical cancer in coastal Karnataka region and HPV variant analysis provide baseline data for prevention and HPV vaccination programs.

Multiple Genetic Marker Analysis with Using Quantitative RT-PCR in Gastric Cancer (위암에서 정량적 역전사 중합효소연쇄반응을 이용한 다중 표지자 분석)

  • Yoo, Moon-Won;Lee, Hyuk-Joon;Choi, Soo-Min;Yu, Ji-Eun;Hur, Keun;Kim, Young-Kook;Yang, Han-Kwang
    • Journal of Gastric Cancer
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    • v.7 no.2
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    • pp.59-66
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    • 2007
  • Purpose: This study was aimed at evaluating the diagnostic validity of peritoneal dissemination of gastric cancer cells by performing multiple genetic marker analysis via quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in gastric cancer cell lines and gastric cancer tissues. Materials and Methods: Quantitative RT-PCR was performed on 12 human gastric cancer cell lines and 10 gastric cancer tissues with four mRNAs of carcinoembryonic antigen (CEA), Cytokeratin 20 (CK20), dopa decarboxylase (DDC), and L-3-phosphoserine phosphatase (L3PP). Results: Out of the 12 human gastric cancer cell lines we tested, CEA was overexpressed in four cell lines (33%), CK20 in one (8%), DDC in six (50%) and L3PP was expessed in all the lines (100%). Out of the 10 gastric cancer tissues we tested, CEA was overexpressed in nine tissues, CK20 in eight, DOC in nine and L3PP was overexpressed in all the tissues. L3PP was overexpressed in all the gastric cancer cell lines and tissues, but the levels of overexpression were lower than those of CEA and DDC. Conclusion: Multiple genetic marker analysis can compensate for the weak points of single marker analysis when testing gastric cancer, and three mRNAs of CEA, DDC and L3PP can be used as candidate genes.

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