• Title/Summary/Keyword: Binding experiment

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The ability of in vitro cultured bovine oviduct epithelial cells in binding and maintaining motility of bull sperm (체외배양 소 난관상피세포의 정자에 대한 결합 및 활력유지능)

  • Roh, Sang-ho;Lee, Byoung-chun;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.220-225
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    • 1999
  • The aim of these experiments was to investigate the effects of bovine oviduct epithelial cells (OEC) derived from different segments to bind sperm binding and maintain their motility in vitro. In experiment 1, the number of sperm attached to OEC derived from isthmus or ampulla, the motility of unattached sperm during co-culture and fertilizing ability were assessed. In experiment 2, heparin treated sperm (hsp) or no treated sperm (nsp) were used to evaluate OEC binding ability of capacitated sperm. In experiment 1, regardless of their origin, approximately 65% of the sperm were attached to OEC within 2h. From 6h of co-culture, the numbers of unattached sperm on ampullary OEC were significantly higher than those on isthmic OEC (p<0.005). From 12h of co-culture, the motility of unattached sperm on isthmic OEC were significantly higher than those on ampullary OEC(p<0.05). The cleavage rate of oocytes inseminated on OEC derived from isthmic segment was also significantly higher than those from ampullary segment (p<0.01). In experiment 2, the numbers of unattached hsp on OEC were significantly higher than those of controls (p<0.01), between 2-24h examination. From 12h of co-culture, the motility of unattached nsp were significantly greater than those of hsp (p<0.01). These results show that bovine OEC derived from the isthmus play more important role(s) for sperm binding, maintaining motility and fertilization in vitro than those from the ampulla, and heparin induced capacitation may change sperm binding ability on OEC in vitro.

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An Experimental Syntactic Study of Korean Anaphor Binding: A case study of 'caki'

  • Kim, Ji-Hye
    • Language and Information
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    • v.15 no.1
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    • pp.63-78
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    • 2011
  • This study investigates the binding behavior of the Korean anaphor 'caki', which has been regarded thus far as a long-distance anaphor (LDA). Given that even local anaphors can be bound long-distance when they function as exempt anaphors in certain languages (Pollard and Sag 1992; Kim and Yoon 2009a, b), I investigated the binding behavior of LD-bound 'caki', in order to determine whether LD-bound 'caki' differs from LD-bound 'caki-casin' in the same contexts. In the experiment, subjects were required to rate the grammaticality of Korean sentences representing various types of LD binding of 'caki' and to determine whether the sloppy or the strict reading was more prominent in elliptical VPs containing the anaphor. The results are discussed with respect to the typology of LDAs proposed by Cole, Hermon and Huang (2001).

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The Experiment Study on Chloride Binding of Cement Paste According to The Al/Ca+Si Ratio (Al/Ca+Si 비에 따른 시멘트 페이스트의 염화물 고정에 관한 실험적 연구)

  • Lee, Yun-Su;Lee, Han-Seung
    • Proceedings of the Korean Institute of Building Construction Conference
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    • 2016.05a
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    • pp.51-52
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    • 2016
  • This paper researches the Chloride Binding of Cement Paste according to the Ca/Si and Ca/Al Ratio. The mechanisms of chloride ion binding are not completely known, although it is believed that Alumina contents in cementitious system have an important role. For changing cement paste composition, Ordinary Portland Cement(OPC) paste is substituted by Granulated Ground Blast Slag(GGBS). With increasing the ratio of GGBS substitution(Thus alumina contents is increasing), The chloride binding capacity has a tendency to increase of binding chloride ion capacity.

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Discrimination of JNK3 bound small molecules by saturation transfer difference NMR experiments

  • Lim, Jong-Soo;Ahn, Hee-Chul
    • Journal of the Korean Magnetic Resonance Society
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    • v.16 no.1
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    • pp.67-77
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    • 2012
  • The small molecule binding to the c-Jun N-terminal kinase 3 (JNK3) was examined by the measurements of saturation transfer difference (STD) NMR experiments. The STD NMR experiment of ATP added to JNK3 clearly showed the binding of the nucleotide to the kinase. The STD NMR spectrum of dNTPs added to JNK3 discriminated the kinase-bound nucleotide from the unbound ones. After the five-fold addition of ATP to the dNTPs and JNK3 mixture, only signals of the cognate substrate of JNK3, ATP, were observed from the STD NMR experiment. These results signify that by the STD NMR the small molecules bound to JNK3 can be discriminated from the pool of the unbound molecules. Furthermore the binding mode of the small molecule to JNK3 can be determined by the competition experiments with ATP.

Characterization of Binding of Treponema denticola to Immobilized Fibrinogen using the Fluorescent Fatty Acid Labeling Method

  • Hong, Jin;Lee, Si-Young
    • International Journal of Oral Biology
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    • v.35 no.3
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    • pp.107-111
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    • 2010
  • Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.

Analysis of Double Stranded DNA-dependent Activities of Deinococcus radiodurans RecA Protein

  • Kim, Jong-Il
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.508-514
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    • 2006
  • In this study, the double-stranded DNA-dependent activities of Deinococcus radiodurans RecA protein (Dr RecA) were characterized. The interactions of the Dr RecA protein with double-stranded DNA were determined, especially dsDNA-dependent ATP hydrolysis by the Dr RecA protein and the DNA strand exchange reaction, in which multiple branch points exist on a single RecA protein-DNA complex. A nucleotide cofactor (ATP or dATP ) was required for the Dr RecA protein binding to duplex DNA. In the presence of dATP, the nucleation step in the binding process occurred more rapidly than in the presence of ATP. Salts inhibited the binding of the Dr RecA protein to double-stranded DNA. Double-stranded DNA-dependent ATPase activities showed a different sensitivity to anion species. Glutamate had only a minimal effect on the double-stranded DNA-dependent ATPase activities, up to a concentration of 0.7 M. In the competition experiment for Dr RecA protein binding, the Dr RecA protein manifested a higher affinity to double-stranded DNA than was observed for single-stranded DNA.

Biological Function of Lactoferrin in Milk

  • Kei-Ichi, Shimazaki
    • 한국유가공학회:학술대회논문집
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    • 2002.04a
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    • pp.37-42
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    • 2002
  • Lactoferrin is an iron-binding glycoprotein and its bacteriostatic and bactericidal effects on Gram-positive and Gram-negative bacteria have been well-known. However, certain kind of lactic acid bacteria are resistant against its antibacterial effects. Moreover, it is reported that lactoferrin promotes the growth of bifidobacteria by in vitro and in vivo experiments. In this experiment, lactoferrin-binding protein was found both in the membrane and cytosolic franctions of Bifidobacterium. Bifidobacterium was grown in anaerobic conditions in MRS broth containing cysteine, gathered by centrifugation and processed by sonication. The lactoferrin-binding proteins on the PVDF-membrane transferred after SDS-PAGE were detected by far-western method using biotinylated lactoferrin and streptavidin-labeled horse radish peroxidase. Observation in growth effects of lactoferrin on Bifidobacterium suggested that there is a relation between the presence of lactoferrin-binding proteins on the cells and their growth.

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Backbone assignment of HMGB1 A-box and molecular interaction with Hoxc9DBD studied by paramagnetic probe

  • Choi, Ji Woong;Park, Sung Jean
    • Journal of the Korean Magnetic Resonance Society
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    • v.25 no.2
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    • pp.17-23
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    • 2021
  • High mobility group protein B1 (HMGB1) is a highly conserved, non-histone, chromatin associated nuclear protein encoded by HMGB1 gene. HMGB1 proteins may be general co-factors in Hox-mediated transcriptional activation that facilitate the access of Hox proteins to specific DNA targets. It is unclear that the exact binding interface of Hoxc9DBD and HMGB1. To identify the interface and binding affinity of Hoxc9DBD and HMGB1 A-box, the paramagnetic probe, MTSL was used in NMR titration experiment. It is attached to the N-terminal end of HMGB1 A-box by reaction with thiol groups. The backbone assignment of HMGB1 A-box was achieved with 3D NMR techinques. The 15N-labeled HMGB1 A-box was titrated with MTSL-labeled Hoxc9DBD respectively. Based on the chemical shift changes we can identify the interacting residues and further map out the binding sites on the protein structure. The NMR titration result showed that the binding interface of HMGB1 A-box is around loop-1 between helix-1 and helix-2. In addition, the additional contacts were found in N- and C-terminus. The N-terminal arm region of Hoxc9DBD is the major binding region and the loop between helix1 and helix2 is the minor binding region.

Metal Sequestering by a Poly(ethylenimine)-Sephadex G-25 Conjugate Containing 2,2'-Dihydroxyazobenzene

  • Gwan, Won Jong;Yu, Chang Eun;Jang, Won Seok;No, Yeong Seok;Seo, Jeong Hun
    • Bulletin of the Korean Chemical Society
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    • v.21 no.4
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    • pp.393-400
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    • 2000
  • 2,2¢-Dihydroxyazobenzene (DHAB) was attached to poly(ethylenimine) (PEI) to obtain DHAB-PEI. Spectral titration revealed that uranyl, Fe(III), Cu(II), and Zn(II) ion form 1 : 1-type complexes with DHAB attached to PEI. Formation constants for the metal complexes formed by the DHAB moieties of DHAB-PEI were mea-sured by using various competing ligands. The results indicated thatthe concentrations of uranyl, Fe(III), and Cu(II) ions can be reduced to 10 -16 -10 -23 M at p 8 with DHAB-PEI when the concentration of the DHAB moiety is 1 residue M. By using cyanuric chloride as the coupling reagent, DHAB-PEI was immobilized on Sephadex G-25 resin to obtain DHAB-PEI-Seph. Binding of uranyl,Fe(III), Cu(II), and Zn(II) ion by DHAB-PEI-Seph was characterized by using competing ligands. A new method has been developed for characteriza-tion of metal sequestering ability of a chelating resin. Formation constants and metal-binding capacity of two sets of binding sites on the resin were estimated for each metal ion. DHAB-PI-Seph was applied to recovery of metals such as uranium,Fe, Cu, Zn, Pb, V, Mn, and W from seawater. The uranium recovery from seawaterby DHAB-PEI-Seph does not meet the criterion for economical feasibility partlydue to interference by Fe and Zn ions. The seawater used in the experiment was contaminated by Fe and Zn and, therefore, the efficiency of uranium extractionfrom seawater with DHAB-PEI-Seph could be improved if the experiment is carried out in a cleaner sea.

Clip-type Binding Implement Effect on Anchorage Behavior of 90-Degree End-Hooked Transverse Reinforcement in Reinforced Concrete Columns (클립형 연결장치로 결속된 90도 갈고리를 갖는 띠철근의 정착거동)

  • Park, Kyoung-Yeon;Yun, Hyun-Do
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.24 no.4
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    • pp.72-80
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    • 2020
  • The purpose of this study is to secure the same or more structural performance and constructability for the details of hooks cross-constructed at 135 degrees used as external-ties standard detail in RC columns, therefore, to the purpose of improving constructability, the clip-type binding implement was suggested and A total of 28 pull-out specimens were prepared with the parameters of concrete compressive strength and clip-embeded length, clip installation location to examine the anchorage behavior of the clip-type binding implement. The experiment was carried out. The results of the experiment confirmed that the anchorage strength of the clip-type binding implement was higher than the details of hooks cross-constructed at 135-degree regardless of the diameter of tie and concrete strength, embeded clip length, clip installation. and The 90-degree end hook with clip-type binding implement was showed a similar an anchorage behavior of 135-degree end-hooked transverse reinforcement, consequently, The 90-degree end hooked with clip-type binding implement is evaluated to be the same anchorage behavior and performance as standard 135-degree end hook detail.