• Title/Summary/Keyword: Specific reactivity

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Fine Mutational Analysis of 2B8 and 3H7 Tag Epitopes with Corresponding Specific Monoclonal Antibodies

  • Kim, Tae-Lim;Cho, Man-Ho;Sangsawang, Kanidta;Bhoo, Seong Hee
    • Molecules and Cells
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    • v.39 no.6
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    • pp.460-467
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    • 2016
  • Bacteriophytochromes are phytochrome-like light-sensing photoreceptors that use biliverdin as a chromophore. To study the biochemical properties of the Deinococcus radiodurans bacteriophytochrome (DrBphP) protein, two anti-DrBphP mouse monoclonal antibodies (2B8 and 3H7) were generated. Their specific epitopes were identified in our previous report. We present here fine epitope mapping of these two antibodies by using truncation and substitution of original epitope sequences in order to identify minimized epitope peptides. The previously reported original epitope sequences for 2B8 and 3H7 were truncated from both sides. Our analysis showed that the minimal peptide sequence lengths for 2B8 and 3H7 antibodies were nine amino acids (RDPLPFFPP) and six amino acids (PGEIEE), respectively. We further characterized these peptides in order to investigate their reactivity after single deletion and single substitution of the original peptides. We found that single-substituted 2B8 epitope (RDPLPAFPP) and dual-substituted 3H7 epitope (PGEIAD) showed significantly increased reactivity. These two antibodies with high reactivity for the short modified peptide sequences are valueble for developing new peptide tags for protein research.

Pathologic observations on the canine mammary gland tumors and immunohistochemical study on the origin of chondroid tissue in mammary gland mixed tumors (개 유선종양의 병리학적 관찰 및 유선 혼합종양의 연골기원에 관한 면역조직화학적 연구)

  • Kim, Yong-baek;Seo, Il-bok;Kim, Jae-hoon;Bak, Eun-jung;Kim, Dae-yong;Han, Jeong-hee
    • Korean Journal of Veterinary Research
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    • v.37 no.4
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    • pp.843-854
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    • 1997
  • Sixteen mammary gland tumors were collected from Seoul National University and Kangwon National University. The average age of the bitches with mammary gland tumor was 10 years. Total 17(60.7%) out of 28 tumor masses observed in 4th and 5th glands. Classification of these tumors according to Hampe and Misdorp were simple adenoma, complex adenoma, benign mixed tumor, papillary adenocarcinoma, solid adenocarcinoma and malignant mixed tumor. Immunohistochemical reaction of the intermediate filaments against normal canine mammary gland showed as followed; anti-cytokeratin 18 was strong and anti-cytokeratin 14 was moderate to the luminal epithelium. Anti-cytokeratin 14 and anti-pancytokeratin to the myoepithelium were showed strong, but anti-vimentin was weak in reactivity. Anti-vimentin to the interstitial cells was represented strong reactivity. The origin of cartilage in mixed tumor of canine mammary gland was studied immunohistochemically with antibodies against intermediate filament. In mammary gland mixed tumors, cartilage tumor tissues were surrounded with the irregularly demarcated three zones composed of adjacent star shaped cells in myxoid areas, proliferative spindle shaped cells and basal located proliferated cells. From basal proliferated cells to star shaped cells, the immunohistochemical reactivity of myoepithelium specific anti-pancytokeratin was decreased gradually and the reactivity of interstitial cell specific anti-vimentin was increased gradually. Based on these immunohistochemical staining patterns, we suggested that the origin of cartilagenous components in canine mammary gland mixed tumor is most likely to the proliferation and metaplsia of myoepithelium.

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FAST REACTOR PHYSICS AND COMPUTATIONAL METHODS

  • Yang, W.S.
    • Nuclear Engineering and Technology
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    • v.44 no.2
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    • pp.177-198
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    • 2012
  • This paper reviews the fast reactor physics and computational methods. The basic reactor physics specific to fast spectrum reactors are briefly reviewed, focused on fissile material breeding and actinide burning. Design implications and reactivity feedback characteristics are compared between breeder and burner reactors. Some discussions are given to the distinct nuclear characteristics of fast reactors that make the assumptions employed in traditional LWR analysis methods not applicable. Reactor physics analysis codes used for the modeling of fast reactor designs in the U.S. are reviewed. This review covers cross-section generation capabilities, whole-core deterministic (diffusion and transport) and Monte Carlo calculation tools, depletion and fuel cycle analysis codes, perturbation theory codes for reactivity coefficient calculation and cross section sensitivity analysis, and uncertainty analysis codes.

An Experimental Study on RCCI(Reactivity Controlled Compression Ignition) Combustion of Dual-fuel due to Injector Characteristics (인젝터 특성에 따른 2중 연료의 RCCI 연소에 관한 실험적 연구)

  • Sung, Ki-An
    • Transactions of the Korean Society of Automotive Engineers
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    • v.20 no.2
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    • pp.110-115
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    • 2012
  • This study describes the characteristics of combustion and exhaust emission in the special engine applying a fuel reactivity controlled compression ignition (RCCI) concept with two different energizing type (solenoid and piezoelectric) injectors for diesel injection. A diesel-gasoline mixed dual-fuel reactivity controlled compression ignition concept is demonstrated as a promising method to achieve high thermal efficiency and low emission in internal combustion engines for transportation vehicles. For investigating the combustion characteristics of RCCI, engine experiments were performed in a light-duty diesel engine over a range of injection timing and mixing rate of gasoline in mass. It was investigated that by increasing the nozzle hole diameter, increasing the combustion pressure and the net indicated mean effective pressure. $NO_x$ and soot can be reduced by advancing start of injection in 84 mixing rate of gasoline in mass. The resulting operation showed that light duty engine could achieve 48 percent net indicated efficiency and 191[g/kW-hr] net indicated specific fuel consumption with lower levels of nitrogen oxides and soot.

Electrophoretic and Immunological Evaluation of Secalin in Rye, Triticale, and Wheat-Rye Translocation Wheat

  • Seo, Yong-Weon;Hong, Byung-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.228-233
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    • 1998
  • Seed storage proteins have been used for studying biochemical genetics and end-use quality aspects. We conducted enzyme-linked immunosorbent assay (ELISA) and one-dimensional SDS-PAGE (1D SDS-PAGE) to evaluate different cereal crop species and Korean wheat lines for rye secalin proteins. The antisecalin antibody showed consistent specificity for rye secalin with little cross-reactivity to gliadins. Immunological cross-reactivities measured by the ELISA technique using competition assay showed significant differences of absorbance among rye, triticale, wheat-rye translocated wheat and non-translocated wheat. The absorbance values were lowest in rye followed by triticale, translocated wheat and non-translocated wheat. The ELISA for discrimination of wheat-rye translocation on the basis of antigen-antibody reactivity showed that none of the Korean wheat lines possessed 1RS and secalin proteins. The competitive ELISA experiment demonstrated specific determination for secalin that was originated from rye chromosomal parts. The result of 1D SDS-PAGE for identifying rye secalin subunits showed all three rye specific secalin protein subunits (75 KDa, 45 KDa, and 40 KDa) for rye and triticale, and 1RS specific secalins (45 KDa and 40 KDa) for 1AL/1RS and 1BL/1RS translocated wheats. All Korean wheats were lacking 1RS of rye chromosome and secalin.

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Production and Characterization of Specific Antibodies to Bombesin

  • Kwon, Hyeok-Yil;Lee, Yun-Lyul;Park, Hyoung-Jin
    • The Korean Journal of Physiology
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    • v.28 no.1
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    • pp.91-97
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    • 1994
  • In order to produce a specific bombesin antiserum far very sensitive radioimmunoassay, synthetic $[lys^3]-bombesin$ conjugated to bovine serum albumin was subcutaneously injected into guinea pigs. The conjugation was performed using either carbodiimide or gIutaraldehyde as a coupling agent. The antisera were characterized by analysis of Scatchard and Sips plots. The antiserum LBE 2G/2 raised by repeat injection of the immunogen conjugated with carbodiimide showed the titer of 1 : 188,000, very low cross-reactivity to bombesin-like peptides except bombesin, with high affinity constant $(1.64{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.91). The antiserum LBG 1G/2 produced by repeat injection of the immunogen conjugated with glutaraldehyde possessed the titer of 1 : 43,000, high cross-reactivity to some bombesin-like peptides, high affinity constant $(1.19{\times}10^{11}\;M^{-1})$ and high heterogeneity index (0.79). These results indicate that the antiserum LBE 2G/2 is specific only to bombesin and that the antiserum LBG IG/2 binds to some bombesin-like peptides such as alytesin, gastrin releasing peptide and neuromedin C. The antiserum LBE 2G/2 is sufficient for the very sensitive radioimmunoassay of bombesin.

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Analysis of the CREOLE experiment on the reactivity temperature coefficient of the UO2 light water moderated lattices using Monte Carlo transport calculations and ENDF/B-VII.1 nuclear data library

  • El Ouahdani, S.;Erradi, L.;Boukhal, H.;Chakir, E.;El Bardouni, T.;Boulaich, Y.;Ahmed, A.
    • Nuclear Engineering and Technology
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    • v.52 no.6
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    • pp.1120-1130
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    • 2020
  • The CREOLE experiment performed In the EOLE critical facility located In the Nuclear Center of CADARACHE - CEA have allowed us to get interesting and complete experimental information on the temperature effects in the light water reactor lattices. To analyze these experiments with accuracy an elaborate calculation scheme using the Monte Carlo method implemented in the MCNP6.1 code and the ENDF/B-VII.1 cross section library has been developed. We have used the ENDF/B-VII.1 data provided with the MCNP6.1.1 version in ACE format and the Makxsf utility to handle the data in the specific temperatures not available in the MCNP6.1.1 original library. The main purpose of this analysis is the qualification of the ENDF/B-VII.1 nuclear data for the prediction of the Reactivity Temperature Coefficient while ensuring the ability of the MCNP6.1 system to model such a complex experiment as CREOLE. We have analyzed the case of UO2 lattice with 1166 ppm of boron in ordinary water moderator in specified temperatures. A detailed comparison of the calculated effective multiplication factors with the reference ones [1] in room temperature presented in this work shows a good agreement demonstrating the validation of our 3D calculation model. The discrepancies between calculations and the differential measurements of the Reactivity Temperature Coefficient for the analyzed configuration are relatively small: the maximum discrepancy doesn't exceed 1,1 pcm/℃. In addition to the analysis of direct differential measurements of the reactivity temperature coefficient performed in the poisoned UO2 lattice configuration, we have also analyzed integral measurements in UO2 clean lattice configuration using equivalency of the integral temperature reactivity worth with the driver core fuel reactivity worth and soluble boron reactivity worth. In this case both of the ENDF/B-VII.1 and JENDL.4 libraries were used in our analysis and the obtained results are very similar.

Characterization and Epitope Mapping of KI-41, a Murine Monoclonal Antibody Specific for the gp41 Envelope Protein of the Human Immunodeficiency Virus-1

  • Shin, Song-Yub;Park, Jung-Hyun;Jang, So-Youn;Lee, Myung-Kyu;Hahm, Kyung-Soo
    • BMB Reports
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    • v.31 no.1
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    • pp.58-63
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    • 1998
  • In this study, a mouse monoclonal antibody (mAb) against gp41(584-618), the immunodominant epitope protein, was generated. For this purpose, BALB/c mice were immunized with double branched multiple antigenic peptides derived from the HIV-1 gp41(584-618) sequence, and antibody-secreting hybridoma were produced by fusion of mice splenocytes with SP2/0 myeloma cells. One clone producing an antigen specific mAb, termed KI-41(isotype IgG1) was identified, whose specific reactivity against gp41(584-618) could be confirmed by ELISA and Western blot analysis. Epitope mapping revealed the recognition site of the mAb KI-41 to be located around the sequence RILAVERYLKDQQLLG, which comprises the N-terminal region within the immunized gp41(584-618) peptied. Since this mAb recognizes this specific epitope within the HIV-1 gp41 without any cross-reactivity to other immunodominant regions in the HIV-2 gp35, KI-41 will provide some alternative possibilities in further applications such as the development of indirect or competitive ELISA for specific antibody detection in HIV-1 infection or for other basic researches regarding the role and function of HIV-1 gp41.

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Development of Monoclonal Antibodies Specific to Galectin of Pine Wood Nematode, Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle and Their Utilization for Detection of Pine Wood Nematodes (소나무재선충[Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle]의 GaLectin에 대한 특이적인 단클론 항체 제작과 진단에의 활용)

  • Kim, A-Young;Kim, Young Ha;Choi, Bo-Hye;Nguyen, Trang;Yoon, Kyungjae Andrew;Lee, Si Hyeock;Han, Hye-Rim;Koh, Young Ho
    • Korean journal of applied entomology
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    • v.57 no.1
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    • pp.7-13
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    • 2018
  • Currently, there is no available tool that rapidly diagnoses pine wood nematode (PWN)-infected pine trees in the field. In this study, we synthesized and purified PWN Galectin, which might be an antigen specific to PWN, using the Baculovirus expression system. We used PWN Galectin as an antigen for generating 1,464 fusion hybridoma cell lines secreting monoclonal antibodies (Mabs). Among them, we selected 62 fusion hybridoma cell lines showing high reactivity to PWN Galectin. We further selected 12 fusion hybridoma cell lines showing high reactivity to the standard PWN-infected pine tree phosphate buffered saline (PBS) extract. Additionally, two fusion hybridoma cell lines showing no or extremely low reactivity were used as controls. The selected fusion hybridoma cell lines were subjected to limiting dilutions for selecting and establishing Mab-secreting cell lines showing higher reactivity to the standard PWN-infected pine tree extract than to the standard normal pine tree PBS extract. Moreover, the selected fusion hybridoma cell lines were further selected based on their higher reactivity to PWN protein extracts than to three non-pathogenic nematode protein extracts. The Mab-secreting cell lines established in this study could be used to develop rapid diagnostic tools that can be used in the field or in laboratories for detecting PWN-infected pine trees or PWN.

Specific Surface Area and Pore Structure Changes of Calcined Lime with Calcination and Sulfation Reaction (소성과 황화반응에 따른 생석회의 비표면적 및 기공구조 변화)

  • 강순국;정명규
    • Journal of environmental and Sanitary engineering
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    • v.13 no.3
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    • pp.19-29
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    • 1998
  • The calcination reactivity of limestone and physical property changes of calcined lime were investigated with a temperature($720~1000^{\circ}C$ under atmospheric gas($N_2$, $CO_2$) conditions. The mechanisms of mass transport in a lime matrix were represented by the evaporation and condensation (${\gamma}=1.7$) at $1000^{\circ}C$ and the volume diffusion (${\gamma}=2.7$) at $800^{\circ}C$, which was obtained by the specific surface area of calcined lime with sintering conditions. Also, the effect of physical property on the reactivity of sulfation reaction was determined by the changes of pore size with $lime-SO_2$ reaction in this work. The initial sulfation rate of calcined lime increased with increasing temperature, whereas the capture capacity of $SO_2$ exhibited a maximum value at $900^{\circ}C$. The pore volume of sulfated lime was decreased with increasing sulfation time, but the major pores shifted to the distribution of larger size at a temperature of $850{\;}~{\;}1000^{\circ}C$. The mean pore size of sulfated lime based on pore volume decreased gradually at $1000^{\circ}C$; however, it increased with sulfation time up to 40 min and rapidly decreased thereafter.

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