• Title/Summary/Keyword: antibody layer

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The comparative study on Newcastle disease virus antibody titer by hemagglutination inhibition test and enzyme-linked immunosorbent assay (혈구응집억제반응과 효소면역측정법을 이용한 닭 뉴캣슬병 바이러스에 대한 혈중항체가 비교)

  • Han, Sung-Tae;Lee, Cheong-San;Kwak, Hak-Koo;Song, Jong-Han;Lee, Jong-In
    • Korean Journal of Veterinary Service
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    • v.26 no.3
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    • pp.215-219
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    • 2003
  • This study was conducted to investigate the similarity between hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay(ELISA) titers and sample to positive ratio (S/P ratio) of Newcastle disease(ND) virus. To perform this study, the 372 sera of broiler chicks and 120 sera of layers and breed chicks were collected from slaughter house and farms, respectively. As a result of HI test out of different chicks, the positive percentage of ND antibody titer of broiler, layer and breeder, when a standard positive HI titer were '2', was 84.4%, 100% and 100%, respectively. The positive percentage of ND antibody titer by ELISA was shown 38.4%, 100% and 100% and S/P ratio were also shown 81.5%, 98.2% and 99.2%, respectively. The results of comparative survey with same sera by two experimental methods were as follows; In low HI titer, ELISA titer was not similar to HI titer, but S/P ratio was similar to it. In high HI titer, ELISA titer was not similar to HI titer. Therefore, HI titer was more similar to S/P ratio than ELISA titer.

Immunochemical Studies of Starfish Gangliosides: Production of Monoclonal Antibody against AG-2, the Major Ganglioside of Starfish Acanthaster planci, and Detecting Its Distribution in Tissues by TLC Immunostaining

  • Miyamoto, Tomofumi;Yamamoto, Atsushi;Sakai, Maki;Tanaka, Hiroyuki;Shoyama, Yukihiro;Higuchi, Ryuichi
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.4
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    • pp.298-304
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    • 2006
  • In this study, we establish a thin-layer chromatography (TLC) immunostaining method for detecting starfish gangliosides. A new monoclonal antibody (MAb) against AG-2, the major gangliosides molecular species of Acanthaster planci, was produced by fusing hybridoma with splenocytes immunized to liposomal AG-2. BALB/c male mice were injected with liposomal AG-2 antigen, and immunized. Their splenocytos were isolated and fused with hypoxanthine-aminopterine-thimidine (HAT)-sensitive mouse myeloma cells. Hybridomas producing MAb reactive to AG-2 were cloned using the limited dilution method. Established hybridomas were cultured in eRDF medium. Crude MAb produced from clone 8D4 was purified with a magnesium pyrophosphate column. Enzyme immunoassay and TLC immunostaining of AG-2 were performed using the purified MAb. Structurally related gangliosides did not cross-react with anti-AG-2 antibodies. The detection limit of TLC immunostaining was 50 ng of AG-2. The newly established immunostaining method was further developed for detecting AG-2 distribution and qualitative analysis in tissues and/or organs. Our results show that the majority of AG-2 is present in the stomach of male A. planci, while AG-2 is distributed not only in the stomach but also in the the pyloric caeca of female A. planci.

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Surface Plasmon Resonance Immunosensor for Detection of Legionella pneumophila

  • Oh, Byung-Keun;Lee, Woochang;Bae, Young-Min;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.112-116
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    • 2003
  • An immunosensor based on surface plasmon resonance (SPR) onto a protein G layer by Self-assembly technique was developed for detection of Legionella pneumophila. The protein G layer by self-assembly technique was fabricated on a gold (Au) surface by adsorbing the 11-mercaptoundecanoic acid (MUA) and an activation process for the chemical binding of the free amino (-NH$_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) in series. The formation of the protein G layer by self-assembly technique on the Au Substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The Surface topographies of the fabricated thin films on an Au substrate were also analyzed by using an atomic force microscope (AFM). Consequently, an immunosensor for the detection of L. pneumophila using SPR was developed with a detection limit of up to 10$^2$CFU per mL.

Prevalence of fowl adenovirus and chicken anemia virus in Jeonbuk, Korea (전북지역 조류아데노바이러스 및 닭전염성빈혈 감염률 조사)

  • Jeong, Han-Sol;Baek, Kui-Jeong;Koh, Won-Seok;Lee, Jeong-Won
    • Korean Journal of Veterinary Service
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    • v.41 no.1
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    • pp.21-27
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    • 2018
  • Fowl adenovirus (FAdV) and chicken anemia virus (CAV) have gained much importance as an immunosuppressive and economically important emerging pathogen of poultry. This study was carried out to investigate the prevalence of FAdV and CAV infection in chickens. The groups were divided into Korean native chickens, broiler, layer hens and broiler breeder and set up groups according to age. As results, 12.5% of the native chicken, 2.5% of broiler and 6.7% of layer chicken were positive, respectively by PCR for FAdV. Serological test showed that 84.8%, 79.0%, 97.7% and 96.1% of chickens were positive for antibody to FAdV in native chickens, broiler, layer hens and broiler breeder. The prevalence of CAV infection were 20.0%, 7.5%, 16.7% and 10.0%, based on CAV gene detection by PCR. In serological test of CAV, 40.6%, 35.9%, 84.8% and 73.9% of chickens were positive in that groups.

The Temporal and Spatial Expression of the Cytokeratin in Keratinocytes during Cutaneous Wound Healing on the Amphibian(Bombina orientalis) (양서류 피부 상처회복과정 중 각질화세포 cytokeratin의 분포)

  • Lim, Do-Seun;Jeong, Soon-Jeong;Moon, Myung-Jin;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.37 no.4
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    • pp.209-217
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    • 2007
  • The system of wound healing is very complex biological processing that includes inflammatory, reepithelialization, and matrix construction. For identification of the transitional pathway of the keratinocytes, we have employed immunohistochemical analysis using cytokeratin antibody after wounding. Epithelium in skin of the frog(Bombina orientalis) was examined with transmission electron microscopy. Cytokeratin was expressed in normal basal and gland cavity cells. At 3-hour basal layer cells were strong positive, however cells of the upper layer were negative reaction. Day1 and 2 after post-wounding, regenerating epithelial cell layer was positive reaction, especially basal layer cells were strong positive. At day 10 after wounding, the degree of positive reaction to basal cells of regenerating epithelial tissue was equal to day 7 wound tissue. At day of 19th, basal and spinous layer cells were strong positive reaction. Regenerating epithelial cells were positive but some basal cells were strong positive at day 27. From this result, we identified that the migration of the keratinocytes in amphibian skin wounds is initiated from basal layer fells and the keratinocytes migrate into basal and middle of the wound area.

The study for the mouse cerebellum developments irradiated with γ-ray during embryogenesis (감마선 조사에 의한 발생과정 중인 마우스의 소뇌발달에 관한 연구)

  • Park, Il-Kwon;Lee, Kyoung-Youl;Park, O-Sung;Kim, Sung-Hwa;Lee, Guen-Jwa;Lee, Kang-Yi;Min, Tae-Sun;Yook, Hong-Sun;Byun, Myung-Woo;Kim, Moo-Kang
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.139-149
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    • 2005
  • In order to study about the lobule and layer formation and cell migration of the mouse cerebellum from at the birth to 15th day effected by 2.5, 5 and 10 Gy r-raddiation at the 19th pregnancy. The routine tissue preparation and staining procedure, Immunohistochemical staining method by the several antibody and western brotting method were utilized from the birth to the15th day. The results were as followings. 1. The body and cerebellum weights were more slowly increase of the the 2.5 Gy, 5 Gy and 10 Gy irradiation group compare to the control group, and the health condition of the 2.5 Gy group was a little bad. but the 10 Gy group was more severe and begun to die from the 12th day after birth. 2. The thickness, proliferation and migration of the 2.5, 5 and 10 Gy irradiated external granular cells from the maginal zone to the medullary area forming the molecular layer from the 6th day to the 15th day after birth were thinner, weaker and more slower according to the radiated dosages than the control group in the cresyl violet staining. 3. The proliteration, migration and lobulation of the 5 Gy radiated groups from the first day to the 15th day after birth were more weak, incomplete and irregular shape in the immunostaining with Dab, Cdk5, P35, calbindin and Zebrin antibody. 4. In the western blotting analysis using the Reelin, Dab, Cdk5 and P35 antibody. The Bands were in the 60 KD, 80 KD, 33 KD and 35 KD, and there were no differences between the control and irradiated groups in the molecular band except the Reelin. 5. As a results, the proliferation and migration of the outer granular and purkinje cells, and lobulation of the cerebellum by the several dosaege of the ${\gamma}$-ray radiation were proportionally incomplete according to dosage.

Ribonucleotide Reductase Activity of Schizosaccbarornyces pombe Is Inhibited by Escherichia coli RecA Antibody (Schizosaccbarornyces pombe에서 Escherichia coli RecA 항체에 의한 Ribonucleotide Reductase 효소활성 저해)

  • Lee, Jung-Sup;Chun, Min-Suck;Kim, Ok-Bong;Park, Jong-Kun;Kim, Si-Wouk;Park, Yeal;Yang, Young-Ki;Hong, Seung-Hwan;Park, Sang-Dai
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.565-569
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    • 1995
  • We have previoosly demonstrated that the RecA-like protein of Schizosaccharomyces pombe (S. pombe) is immunologically related to Escherichia coil (E. coil) RecA protein and that the cellular level of the protein is significantly increased by inhibitors of nucleotide pool-forming enzymes such as hydroxyurea (HU) and methotrexate (MTX) (lee and Park, 1994; lee et al., 1994). In this study, we report that the ribonudeotide redudase activity of S. pombe is inhibited by E. coil RecA antibody, as determined by thin layer chromatography using [5-$^3$H]CDP as a substrate. The relative activity of ribonucleotide reductase was dramatically inhibited by 100 mM of flu (26.4% reduction) in in vitro assay, compared to that of non-treated control. The ribonucleotide reductase activity was also inhibited by immunoprecipitation with E. coil RecA antibody (43.3% reduction). These results indicate that the strudure of S. pombe ribonucleotide reductase is in part similar to that of E. coil RecA protein.

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Calretinin-Containing Neurons in the Deeper Layers of the Hamster Superior Colliculus (햄스터 상구의 deeper layers에서 calretinin이 함유 신경세포)

  • Kim, Ye-Eun;Choi, Jae-Sik;Kim, Hye-Hyun;Yeo, Jin-Yeon;Jeon, Chang-Jin
    • Journal of Life Science
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    • v.16 no.5
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    • pp.750-758
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    • 2006
  • Calcium-binding protein calretinin is thought to play important roles in calcium buffering. Recently, we reported on the distribution, morphology of calretinin-immunoreactive (IR) neurons and the effects of eye enucleation on the immunoreactivity of calretinin in the superficial layers of the hamster superior colliculus (SC). In the present study, we describe the distributions and types of labeled cells and effects of enucleation in the deeper layers by immunocytochemistry. We also compare this labeling to that of GABA, the major inhibitory neurotransmitter in the central nervous system. In contrast to the superficial layers, the deeper layers contained many calretinin-IR neurons which formed two tiers. The first tier, which was very distinctive, was found within the intermediate gray layer. The second tier was found in the deep gray layer. Labeled neurons varied dramatically in morphology and included vertical fusiform, stellate, round/oval, and horizontal neurons. In contrast to the superficial layers, enucleation appeared to have no effect on the distribution of calretinin immunoreactivity in the deeper layers. Two-color immunofluorescence revealed that none of calretinin-IR neurons were labeled with an antibody to GABA. The present results demonstrate that calretinin identifies unique neuronal sublaminar organizations in the hamster SC. The present results also demonstrate that none of the calretinin-IR neurons in the hamster SC is GABAergic interneurons. As many calretinin-IR cells are GABAergic interneurons in most other brain areas, this phenomenon in hamster SC is exceptional.

Isolation of Salmonella from the layer chickens reacting in pullorum-typhoid agglutination test (추백리 혈청검사 양성 산란계로부터 Salmonella속균 분리)

  • 류재윤;전무형;장경수;손현수;곽학구;박경재;우용구
    • Korean Journal of Veterinary Service
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    • v.22 no.3
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    • pp.221-237
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    • 1999
  • To investigate the specificity of rapid slide agglutination test for pullorum-gallinarum diseases and to obtain a basic data for avian salmonellosis control, salmonella isolation was peformed for the layer chickens positively reacted in pullonlm-typhoid agglutination test. The biochemical, serological and antimicrobial properties of the isolates were examined. The results obtained through this study were summarized as follows; 1. Of 2,384 chickens tested by the agglutination test, 606 chickens (25.4%) were positive reactors. 154 of 606 reactors and 49 of the non-reacting chickens were investigated for salmonella isolation, resulting in isolation of 68 strains of salmonellae from 27 chickens. 2. By organs, the isolation frequency from liver, cecum, spleen, ovary and gall bladder showed 8.9% (18 strains), 8.9% (18 strains), 7.4% (15 strains), 4.4% (9 strains) and 3.9% (8 strains), respectively. 3. By culture medium the combination of selenite broth and MacConkey agar revealed the highest isolation rate and the enrichment culture by delayed secondary enrichment culture method was found the most effective for salmonella isolation. 4. The serotypes of 68 salmonella isolates were identified as 3 strains of S pullorum, 24 strains of S gallinarum, 15 strains of S typhimurium, 8 strains of S enteritidis, 7 strains of S paratyphi A, 5 strains of S typhimurium and 6 strains of the other salmonellae. 5. The serotypes of 8 salmonella strains isolated from 49 chickens non-reacting in pullorum-typhoid agglutination test were identified as 3 strains of S typhimurium and 5 strains of S infantis. 6. When 24 chickens of which 68 strains of salmonellae isolated were examined by microplate agglutination test, the average antibody titer for pullorum antigen was $2^{5.25}$. The chickens at antibody titer between $2^3$ and $2^5$ showed the higher frequency of isolation as compared with the chickens at the other titers. 7. When salmonella isolates were tested the antimicrobial drug sensitivity by disk diffusion method, S paratyphi A were highly sensitive by 100% to ATM and GM, S typhimurium, by 88% to AM, CIP, IMP and TN, S infantis, by 100% to AM, CRO, ENR and PIP, S enteritidis,by 100% to IMP and PIP, S pullorum, by 100% to ATM, CRO, ENR and PIP and S gallinarum, by 92% to CRO, CIP and PIP.

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Characterization of a Lactobacillus acidophilus Strain Isolated from Korean Infant Feces and Cloning of Surface Layer Protein Gene slp and Its Expression in Escherichia coli (유아 분변에서 분리한 Lactobacillus acidophilus의 특성 및 표면 단백질 유전자 클로닝과 대장균 내에서의 발현)

  • Park, Myeong-Soo;Ji, Geun-Eog;You, Kwan-Hee;Lee, Si-Kyung;Jeong, Won-Seok;Kim, Jin-Hyung;Jo, Myoung-Hwan;Kim, Soo-Young
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.352-356
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    • 2007
  • A Lactobacillus sp. has been isolated from infant feces and characterized according to its physiological properties and identified as Lactobacillus acidophilus KLA1012. A gene coding surface layer protein (SLP) has been cloned and the sequence has been determined. The nucleotide sequence of slpA was 1,338 bp in size and was identical to that of L. acidophilus ATCC 4356 (100%). Amino acid sequence of SLP-A was deduced from the nucleotide sequence and it had signal sequence at N-terminal, consisting of positively charged amino acid mainly lysine. slpA was cloned and heterologously expressed in E. coli M15 and the 45.2 kDa surface-layer protein band was examined by SDS-PAGE and confirmed by Western blotting using polyclonal antibody against L. acidophilus KLA 1012 SLP-A protein.