• Title/Summary/Keyword: cELISA

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Population of Rice Stripe Virus-Viruliferous Insect and Natural Weed Host of Rice Stripe Virus.

  • Park, Jin-Woo;Jin, Tae-sung;Shin, Dong-bum;Park, Byung-ryul;Kim, Jin-young;Oh, In-suk;Lee, B. C.;T. H. Noh;S. J. Ko
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.140.2-141
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    • 2003
  • Among over-wintering small brown planthoppers, population of the rice stripe virus (RSV)-viruliferous insects was surveyed throughout the country in late April of 2003 by using DAS-ELISA. Averaged population of the RSV-viruliferous insects in this year was 2.1%, which was lower than that of last year of 3.7%. However, the insect population in Seoul, Incheon and Kyeonggi areas were relatively high showing 6.7%, 6.2% and 2.6%, respectively. Based on the survey results, it was expected that overall occurrence of RSV on rice could be decreased in this year, except certain areas. Ovarial transmission rate of RSV by the insects on diseased rice samples collected from 10 areas ranged from 22.2% to 77.8%. Among 35 graminous weed species collected from rice fields in Ganghwa and Kimpo in 2002 and 2003, common reed and formosens were found to be infected by RSV. The result indicates that those weeds are potential alternative natural hosts of the RSV Further studies on ecological and pathological impacts of the alternative natural host of RSV are being processed.

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Isolation and Characterization of Single-Chain Fv Against Ductal Cells

  • Lee Myung-Hoon;Ryu Hye-Myung;Kim Sun-Zoo;Park Ji-Young;Uhm Ji-Hyun;Park Tae-In
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.211-217
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    • 2004
  • For discrimination of ductal and ascinar cells, we isolated a single-chain variable domain fragment (scFv) antibody against ductal cells of salivary gland using phage display technique. From the spleen of a mouse immunized with ductal cell lysate, total RNA was prepared and used as a template for cDNA synthesis of antibody genes. The scFv genes were constructed with variable domain genes of heavy and light chain and were introduced into pCANTAB5E to construct phage scFv library. The phage particles specific for acinar cells were screened by subtraction using immunotubes coated with acinar and ductal cell lysate and enzyme-linked immunoabsorbance assay (ELISA). The characteristics of the scFv were determined by immunohistochemistry (IHC) and the result indicated that the isolated scFv has the specificity against ductal cells of salivary glands and tubules of kidney. And the scFv has an unique binding activity specific for Hashimoto's thyroiditis. The nucleotide sequence of isolated scFv gene was determined and revealed that V/sub H/ belongs to the mouse H-chain family subgroup IB and V/sub L/ to the mouse L-chain family subgroup III.

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The Anti-Depressive Effects of BanHaHuBakTang-kami (BHHBT) after Chronic Immobilization Stress in C57BL/6 Mice (우울증 생쥐 모델에서 반하후박탕가미(半夏厚朴湯加味)의 항우울 효과)

  • Kim, Kook-Ki;Lee, Sang-Ryong;Jung, In-Chul
    • Journal of Oriental Neuropsychiatry
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    • v.25 no.2
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    • pp.191-202
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    • 2014
  • Objectives: The purpose of this study was to examine the anti-depressive effects of BanHaHuBakTang-kami (BHHBT) on an animal model of depression induced by chronic immobility stress. Methods: Mice were treated daily with immobilization stress for 2 hours over a period of 21 days. To examine the effect of BHHBT, we performed behavioral, biochemical and histological analysis to measure immobility time (FST), brain neurotransmitter concentration (HPLC, ELISA), hippocampal damage (H&E staining) and CRF-R1 expression (immunohistochemistry). Results: BHHBT has reduced the immobility time of immobilization stress exposed mice in the forced swimming test. BHHBT has increased the amount of serotonin in the brain. BHHBT has increased the expression level of serotonin in the brain. BHHBT 540 mg/kg were sufficient to prevent tissue damage in the hippocampus region. BHHBT has reduced the expression level of CRF receptors in the hippocampus region. Conclusions: These results suggest that BHHBT may have anti-depressive effects on mice treated with immobilization stress by reducing immobility, increasing brain serotonin concentration and reducing CRF-R1 expression in the hippocampus region.

Detection of Plant RNA Viruses by Hybridization Using In Vitro Transcribed RNA Probes (In Viro 전사 RNA Probe를 이용한 식물 바이러스병의 진단)

  • 최장경;이종희;함영일
    • Korean Journal Plant Pathology
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    • v.11 no.4
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    • pp.367-373
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    • 1995
  • The cDNAs derived from the coat protein (CP) genes of six plant RNA viruses, tobacco mosaic virus-pepper strains (TMV-P) and -ordinary strain (TMV-OM), potato virus Y (PVY), turnip mosaic virus (TuMV), cucumber mosaic virus (CMV) and potato leafroll virus (PLRV), were subcloned into the transcription vector, pSPT18, containing SP6 and T7 promoters. The digoxigenin (DIG)-labeled RNA polymerase after linearlization of the cloned pSPTs with XbaI or SacI, and were tested for their sensitivities for the detection of the six viruses. In slot-blot hybridization, dilution end points for the detection of TMV-P and TMV-OM were 10-4, while those of PVY, TuMV and CMV were 10-3. PLRV was detected at the dilution of 10-2. When each RNA probe was applied for the detection of the viruses in the preparations from the leaf disks (8 mm in diameter, and 12 to 15 mg in weight) of infected natural host plants, TMV-P, TMV-OM and TuMV could be detected from one disk, while PVY from 1 or 2 disks. CMV was detected in the preparation from two disks, and PLRV from three disks. With DIG-labeled RNA probe, PVY was detected at 5 days after inoculation, but with ELISA the virus was detected at 8 days after inoculation to tobacco (Nicotiana tabacum cv. Xanthi nc) plants on which symptoms appeared at 9 days after inoculation. No difference was observed in cross reaction between the RNA probes for the detection of TMV-P and TMV-OM.

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In Vitro Expression of the Recombinant hFSH Gene using Retrovirus Vector System (In Vitro에서 Retrovirus Vector System을 이용한 재조합 hFSH 유전자의 발현)

  • Min, Gyeong-Heon;Kwon, Mo-Sun;Kim, Teoan;Koo, Bon-Chul
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.115-121
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    • 2011
  • hFSH is a glycoprotein secreted from anterior pituitary and consists of ${\alpha}$ and ${\beta}$ subunits. Because of its major biological functions including sperm formation in the male and for follicular growth, FSH is used to cure woman's sterility. In this study we tried to produce recombinant hFSH in vitro using a retrovirus expression vector. Two major components of the vector we constructed are: ( i ) a DNA fragment containing ${\alpha}$ and ${\beta}$ genes fused by a DNA sequence coding carboxyl terminal peptide (CTP) of human chorionic gonadotropin, (ii) a DNA fragment corresponding woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). Evaluation of expression profile of the recombinant FSH using reverse transcription PCR and enzyme-linked immunosorbent assay (ELISA). Among three cell lines tested, HeLa cells were the best for hFSH expression (5,395 mIU/ml), then followed by chicken embryonic fibroblast (CEF) cells and Chinese hamster ovary (CHO) cells in the order of hFSH production. In addition to the amount, the FSH produced from HeLa cells was highest in terms of biological activity which was determined by measuring cAMP.

Surveillance for Equine Infectious Anemia in Jeju Island (제주 사육 말에서 말전염성빈혈 조사)

  • Yang, Jaehyuk;Lim, Yoon-Kyu
    • Journal of Veterinary Clinics
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    • v.31 no.4
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    • pp.357-359
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    • 2014
  • Equine infectious anemia (EIA) is a worldwide infectious disease of horses and other equids. The large serological survey of EIA was performed in Jeju from 2005 through 2011. Using the conventional enzyme-linked immunosorbent assay (cELISA), a total of 10,040 animals (1,329 Jeju Ponies, 8,324 Jeju Pony-Crossbreds and 387 Thoroughbred horses) was tested at the Equine Hospital of Jeju Race Park or Jeju Stud Farm, Korea Racing Authority. This survey found no serological evidence of EIA presence in Jeju. There had been no horse and pony with antibody against EIA since 1985 and no official report on outbreak the disease. Therefore, surveillance conducted found no evidence of EIA activity in Jeju.

Production of monoclonal antibody(MAb) for lymphocytes of Korean native cattle 2. MAb to E-rosette forming lymphocytes. (한우 림프구에 대한 단일항체 생산 2. E-rosette형성 림프구에 대한 단일항체)

  • Song, Hee-jong;Woon, Jae-ho;Kim, Tae-joong
    • Korean Journal of Veterinary Research
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    • v.35 no.4
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    • pp.777-783
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    • 1995
  • A monoclonal antibody(MAb), KCT-23ER, with specificity for E-rosetted T cells of Korean native cattle was prepared by cell hybridization of myeloma P3/NS-1/1-Ag-4-1 and spleen cells from BALB/c mice hyperimmunized with E-rosetted lymphocytes. The isotype of KCT-23ER to T lymphocytes was mouse $IgG_{2b}$. KCT-23ER was reacted with 53.6% to peripheral blood lymphocytes and with 67.8% to nylon wool nonadherent blood lymphocytes. And it was reacted with 72.2%, 59.2% and 35.3% to thymocytes, prescapular lymph node cells and splenocytes, respectively. Immunocytological reactive rates to E-rosetted and non-E-rosetted cells were 72.5% and 22.4%, respectively. These results indicated that KCT-23ER reacted to E-rosetted cells was one of the MAb for investigate of $CD_2$ receptor positive cell subset in the Korean native cattle.

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Production of monoclonal antibody(MAb) for lymphocytes of Korean native cattle 1. MAb to T lymphocytes (한우 림프구에 대한 단일항체 생산 1. T 림프구에 대한 단일항체)

  • Song, Hee-jong;Woon, Jae-ho;Kim, Tae-joong;Chai, Hyo-seok;Hur, Boo-hong;Choi, Min-soon
    • Korean Journal of Veterinary Research
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    • v.35 no.4
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    • pp.769-776
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    • 1995
  • This study was undertaken to develop the monoclonal antibody(MAb) for lymphocytes of Korean native cattle by the cell hybridization of myeloma P3/NS-1/ 1-Ag-4-1 and spleen cells from BALB/c mice hyperimmunized with nylon wool column eluted peripheral T lymphocytes of Korean native cattle. The isotype of MAb KCT-14 against T lymphocyte was mouse $IgG_1$. KCT-14 positivity of mononuclear cells(MNC) from peripheral blood lymphocytes, nylon wool nonadherent and adherent-lymphocytes was 41.7%, 58.4% and 22.6%, respectively. And that of mesenteric lymph node-, spleen and thymus-MNC was 43.3%, 40.2% and 33.6%, respectively. Immunoperoxidase staining of frozen tissue sections showed that the MAb positive cells were located in the medulla of the thymus and in the paracortical area and the mantle zone of the germinal center in the lymph nodes. These results indicated that KCT-14 was one of the MAb for investigate of T lymphocyte subpopulations in the Korean native cattle.

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Characterization of Mouse Hybridoma Producing Monoclonal Antibody Paragonimus westermani (폐디스토마에 대한 Mmonoclonal Antibody생산 세포주개발에 대한 연구)

  • 고광삼;이숙영;이근배
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.64-68
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    • 1985
  • In this study, hybridoma techniques were applied to produce monoclonal antobodies to Paragonimus westermani, commonly known as lung distoma. Balb/c mice were immunized intraperitoneally every week with increasing doses of purfied protein of Paragonimus westermani adult worms begining with 0.3/mg/mouse/7 days and ending with 0.5mg at 28th day. Spleen cells from these immunized mice were hybridized with myeloma cells (NS-1) and the hybridized cells were selected in HAT media. The antibody secreting cells among the hybrid cells were initially selected by ELISA. Those initially selected cells were further screened by the criteria of antibody producing activity, and seneral cell lines among them were further tested with immunodiffusion method. One hybridoma clone produced IgM and another clone produced IgG1. The supernatant of the hybridoma clone producing IgM had titer 1:64 and the hybridoma clone producing IgG1 had titer 1:256 measured by immunofluorescence technique.

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ADSORPTION OF HUMAN RECOMBINANT FIBRONECTIN TO TITANIUM IN VITRO

  • Kim H.J.;Chang I.T.;Koak J.Y.;Heo S.J.;Yim S.A.;Jang J.H.
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.3
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    • pp.301-306
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    • 2004
  • Statement of problem. Fibronectin mediates its biological effects by binding to integrins on cell membranes through a consensus site including the Arg-Gly-Asp (RGD) sequence within tenth type III module. Purpose. The purpose of our study was to investigate the adsorption affinity of human recombinant fibronectin peptide (hFNIII 9-10) to titanium and to investigate the effect of the surrounding ionic composition on the adsorption process. Material and methods. As for evaluating the affinity of hFNIII 9-10 to Ti, titanium disks were incubated in 40, 80 and $120{\mu}g/ml$ hFNIII 9-10 solution at $37^{\circ}C$ overnight, repectively. As for evaluating the effect of surrounding ionic concentration, hFNIII 9-10 was dissolved in distilled water, phosphate buffered saline and RPMI 1640. Optical density (O.D.) was measured in ELISA reader. Results. The results were as follows; 1. The adsorption of hFNIII 9-10 showed significantly highest mean optical density (O.D.) value in $80{\mu}g/ml$. 2. The difference of ionic composition in DW, PBS and RPMI did not influence the adsorption amount of hFNIII 9-10.