• Title/Summary/Keyword: Domain-specificity

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Ectopic Overexpression of Coiled-Coil Domain Containing 110 Delays G2/M Entry in U2-OS Cells

  • Lee, Sue Nyoung;Hong, Kyeong-Man;Seong, Yeon Sun;Kwak, Sahng-June
    • Development and Reproduction
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    • v.24 no.2
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    • pp.101-111
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    • 2020
  • Coiled-coil domain containing 110 (CCDC110, KM-HN-1) is a protein containing C-terminal coiled-coil domain (CCD) which was previously discovered as a member of the human cancer/testis antigen (CTA). In addition, CCDC110 has both nuclear localization signal sequence and the leucine zipper motif. Although the functional role of CCDC110 has yet to be fully identified, the mRNA expression levels of CCDC110 are known to be highly elevated in various cancer types including testis, implying its relevance to cancer pathogenesis. In this study, we first developed several monoclonal antibody (mAb) hybridoma clones targeting CCDC110 and further isolated clone by characterizing for its specificity using immunoblotting and immunoprecipitation approaches with basal parenchymal sperm cells in testis tissue. Next, using these mAbs, we showed that the Tet-inducible overexpression of CCDC110 protein delayed the entry of G2/M phase in U2-OS osteosarcoma cells. Based on these results, we propose that CCDC110 plays a crucial role in cell cycle progression.

Atrial Fibrillation Detection Algorithm through Non-Linear Analysis of Irregular RR Interval Rhythm (불규칙 RR 간격 리듬의 비선형적 특성 분석을 통한 심방세동 검출 알고리즘)

  • Cho, Ik-Sung;Kwon, Hyeog-Soong
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.15 no.12
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    • pp.2655-2663
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    • 2011
  • Several algorithms have been developed to detect AF which rely either on the form of P waves or the based on the time frequency domain analysis of RR variability. However, locating the P wave fiducial point is very difficult because of the low amplitude of the P wave and the corruption by noise. Also, the time frequency domain analysis of RR variability has disadvantage to get the details of irregular RR interval rhythm. In this study, we describe an atrial fibrillation detection algorithm through non-linear analysis of irregular RR interval rhythm based on the variability, randomness and complexity. We employ a new statistical techniques root mean squares of successive differences(RMSSD), turning points ratio(TPR) and sample entropy(SpEn). The detection algorithm was tested using the optimal threshold on two databases, namely the MIT-BIH Atrial Fibrillation Database and the Arrhythmia Database. We have achieved a high sensitivity(Se:94.5%), specificity(Sp:96.2%) and Se(89.8%), Sp(89.62%) respectively.

Exploration of Creativity Construction Components in Computer Science (정보과학 창의성의 구성 요소 탐색)

  • Yun, Seon-Hee;Kim, Yung-Sik
    • The Journal of Korean Association of Computer Education
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    • v.14 no.1
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    • pp.45-54
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    • 2011
  • Creativity is a fine ability among major values of human resource in knowledge-based society. The purpose of this study is to identify the components of the informatics creativity based on domain-specificity and confluence theory. To explore these components, a three-round, classical Delphi process was administered with an expert panel of 31 individuals. All members of the expert panel initially responded to open-ended questions, and the qualitative results were analyzed. The subsequent two rounds of the Delphi required quantitative responses in which the expert panel was asked to indicate the level of importance using a five point Likert scale associated with specific items. As a result of the Delphi survey, total of 32 items were selected as three core areas such as congnitive, affective and environmental components of informatics creativity.

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Molecular Cloning and Substrate Specificity of Human NeuAc ${\alpha}$2,3Gal${\beta}$ 1,3GalNAc GalNac ${\alpha}$2,6-Sialyltransferase (hST6GalNac IV)

  • Lee, Young-Choon;Kim, Kyoung-Sook;Kim, Sang-Wan;Min, Kwan-Sik;Kim, Cheorl-Ho;Choo, Young-Kug
    • Journal of Life Science
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    • v.11 no.1
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    • pp.57-64
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    • 2001
  • The cDNA encoding human NeuAc ${\alpha}$2,3Gal$\beta$ 1,3GalNAc GalNac ${\alpha}$2,6-Sialyltransferase (hST6GalNac IV) was isolated by screening of human fetal liver cDNA library with a DNA probe generated from the cDNA sequence of mouse ST6Gal NAc IV (mkST6GalNAc IV). The cDNA sequence included an open reading frame coding for 302 amino acids, and comparative analysis of this cDNA with mST6GalNAc IV showed that each sequence of the predicted coding region contains 88% and 85% identifies in nucleotide and amino acid levels, respecively. The primary structure of this enzyme suggested a putative domain structure, like that in other glycosyltransferases, consisting of a short N-terminal cytoplamic domain, a transmembrane domain and a large C-terminal active domain. This enzyme expressed in COS-7 cells echibited transferase activity toward NeuAc ${\alpha}$2,3Gal$\beta$ 1,3GalNAc, fetuin and GM1b, although the activity toward the later is very low, no significant activity being detected toward Gal${\beta}$ 1,3Gal NAc or asialofetuin, the other glycoprotein substrates tested. The $^{14}$ C-sialylated residue of fetuin sialylated by this enzyem with CMP-[$^{14}$C]NeuAc was sensitive to treatment with ${\alpha}$2,8-specific sialidase of Vibrio cholerae but resistant to treatment with ${\alpha}$2,3-specific sialidase (NaNase I), and ${\alpha}$2,3- and ${\alpha}$2,8-specific sialidase of Newcastle disease virus. These results clearly indicated that the expressed enzyme is a type of GalNAc ${\alpha}$2,6-sialyltransferase like mST6GalNAc IV, which requires sialic acid residues linked to Gal${\beta}$1,3GalNAc-residues for its activity.

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Selection and identification of single-domain antibody against Peste des Petits Ruminants virus

  • Liu, Dan;Li, Lingxia;Cao, Xiaoan;Wu, Jinyan;Du, Guoyu;Shang, Youjun
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.45.1-45.13
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    • 2021
  • Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

A Study on the Role of Collaboration as Safeguard for Supplier's Relationship-specific Intangible Asset (공급자가 구축한 관계 특유적 무형자산의 보호수단으로서 협업의 역할에 관한 연구)

  • Kim, Jin Wan;Lee, Seok Yong;Park, Ji Young;Hong, Tae Ho
    • Knowledge Management Research
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    • v.10 no.3
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    • pp.35-49
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    • 2009
  • The purpose of this study is to examine whether suppliers are able to benefit by means of constructing a relationship-specific intangible asset and participating in collaboration with purchasers to actively maintain value created while mutually using inter-organizational information systems. To achieve the purpose of this study. 82 questionnaires have been analyzed from suppliers which are using electronic transactions through inter-organizational information systems. As a result, business process specificity and domain knowledge specificity, which are relationship-specific intangible assets, affect operational collaboration and strategic collaboration, which are classified by level of decision-making. Although both types of collaboration are needed to achieve both strategic and operational benefits, we find each collaboration to be uniquely promoted by a specific types of relationship-specific intangible asset. Operational collaboration is found to be an antecedent to operational benefit and strategic collaboration is found to be an antecedent to strategic benefits. No crossover between collaboration and supplier's benefits is found. Consequently, the result of this study shows collaboration is needed for domestic suppliers, which have relatively low levels of collaboration, to keep maintain relationship-specific intangible assets and to prevent the relationship termination cost when transactions have been stopped between supplier and purchaser. Also the results imply the supplier can have advantages by participating in collaboration.

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Distinctive pH Dependence and Substrate Specificity of Peptide Hydrolysis by Human Stromelysin-1 (Stromelysin-1에 의한 펩타이드 가수분해에서 pH와 기질특이성 연구)

  • ;Marianne V. Sorensen
    • Journal of Life Science
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    • v.10 no.2
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    • pp.210-217
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    • 2000
  • A kinetic profile of the catalytic domain of stromelysin-1 (SCD) using the fluorescent peptide substrate has been determined by the stopped-flow technique. The pH profile has a pH optimum of about 5.5 with an extended shoulder above pH 7. Three pKa values, 5.0, 5.7, and 9.8 are found for the free enzyme state and two pH independent Kcat/Km values of 4.1$\times$104 M-1 s-1 and 1.4$\times$104 M-1 s-1 at low and high pH, respectively. The profile is quite different in shape with other MMP family which has been reported, having broad pH optimum with two pKa values. The substrate specificity of SCD towards fluorescent heptapeptide substrates has been also examined by thin layer chromatography. The cleavage sites of the substrates have been identified using reverse-phase HPLC method.SCD cleaves Dns-PLA↓L↓WAR and Dns-PLA↓L↓FAR at two positions. However, the Dns-PLA↓LRAR, Dns-PLE↓LFAR, adn Dns-PLSar↓LFAR are cleaved exclusively at one bond. The double cleavages of Dns-PLALWAR and Dns-PLALFAR by SCD are in marked contrast to the close structurally related matrilysin. A notable feature of SCD catalysis agrees with the structural data that the S1' pocket of SCD is deeper than that of matriysin. The differences observed between SCD and matrilysin may form the basis of understanding the structural relationships and substrate specificities of the MMP family in vivo.

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Diagnostic Efficacy of a Recombinant Cysteine Protease of Spirometra erinacei Larvae for Serodiagnosis of Sparganosis

  • Rahman, S.M. Mazidur;Kim, Jae-Hwan;Hong, Sung-Tae;Choi, Min-Ho
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.41-46
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    • 2014
  • The mature domain of a cysteine protease of Spirometra erinacei plerocercoid larva (i.e., sparganum) was expressed in Escherichia coli, and its value as an antigen for the serodiagnosis of sparganosis was investigated. The recombinant protein (rSepCp-1) has the molecular weight of 23.4 kDa, and strongly reacted with the sparganum positive human or mice sera but not with negative sera by immunoblotting. ELISA with rSepCp-1 protein or sparganum crude antigen (SeC) was evaluated for the serodiagnosis of sparganosis using patient's sera. The sensitivity and specificity of ELISA using rSepCp-1 protein were 95.0% (19/20) and 99.1% (111/112), respectively. In contrast, the sensitivity and specificity of ELISA with SeC were 100% (20/20) and 96.4% (108/112), respectively. Moreover, in experimentally infected mice, the sensitivity and specificity of both ELISA assays were 100% for the detection of anti-sparganum IgG. It is suggested that the rSepCp-1 protein-based ELISA could provide a highly sensitive and specific assay for the diagnosis of sparganosis.

Regulation of signal transducer and activator of transcription 3 activation by dual-specificity phosphatase 3

  • Kim, Ba Reum;Ha, Jain;Kang, Eunjeong;Cho, Sayeon
    • BMB Reports
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    • v.53 no.6
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    • pp.335-340
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    • 2020
  • Since cancer is the leading cause of death worldwide, there is an urgent need to understand the mechanisms underlying cancer progression and the development of cancer inhibitors. Signal transducer and activator of transcription 3 (STAT3) is a major transcription factor that regulates the proliferation and survival of various cancer cells. Here, dual-specificity phosphatase 3 (DUSP3) was identified as a regulator of STAT3 based on an interaction screening performed using the protein tyrosine phosphatase library. DUSP3 interacted with the C-terminal domain of STAT3 and dephosphorylated p-Y705 of STAT3. In vitro dephosphorylation assay revealed that DUSP3 directly dephosphorylated p-STAT3. The suppressive effects of DUSP3 on STAT3 were evaluated by a decreased STAT3-specific promoter activity, which in turn reduced the expression of the downstream target genes of STAT3. In summary, DUSP3 downregulated the transcriptional activity of STAT3 via dephosphorylation at Y705 and also suppressed the migratory activity of cancer cells. This study demonstrated that DUSP3 inhibits interleukin 6 (IL-6)/STAT3 signaling and is expected to regulate cancer development. Novel functions of DUSP3 discovered in IL-6/STAT3 signaling regulation would help expand the understanding of cancer development mechanisms.

A Visualization and Inference System for Protein-Protein Interaction (단백질 상호작용 추론 및 가시화 시스템)

  • Lee Mi-Kyung;Kim Ki-Bong
    • Journal of KIISE:Software and Applications
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    • v.31 no.12
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    • pp.1602-1610
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    • 2004
  • As various genome projects have produced enormous amount of biosequence data, functional sequence analysis in terms of tile nucleic acid and protein becomes very significant. In functional genomics and proteomics, the functional analysis of each individual gene and protein remains a big challenge. Contrary to traditional studies, which regard proteins as not components of a whole protein interaction network but individual entities, recent studies have focused on examining functions and roles of each individual gene and protein in view of a whole life system. In this regard, it has been recognized as an appropriate method to analyze protein function on the basis of synthetic information of its interaction and domain modularity. In this context, this paper introduces the PIVS (Protein-protein interaction Inference & Visualization System), which predicts the interaction relationship of input proteins by taking advantage of information on homology degree, domain modules which input sequences contain, and protein interaction relationship. The information on domain modules can increase the accuracy of the function and interaction relationship analysis in terms of the specificity and sensitivity.