• Title/Summary/Keyword: cELISA

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Characterizations of Tobacco Mosaic Virus isolated from Chinese Foxglove(Rehmannia glutinosa Libosch) (지황(Rehmannia glutinosa Libosch)에서 분리한 Tobacco Mosaic Virus의 특성)

  • 박준식;최민경;유강열;이귀재
    • Korean Journal of Plant Resources
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    • v.16 no.3
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    • pp.230-237
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    • 2003
  • This study was conducted to investigate the occurrence and characterization of tobacco mosaic virus(TMV) in Chinese foxglove isolated from the field of the Chonbuk province(Jinan, Jangsu, Jeongeup). TMV was detected in all three regions and confirmed positive reaction by ELISA test. In the host range test, Chenopodium amaranticola, Nicotiana glutinosa, N. tabacum cv. 'Bright yellow', N. tabacum cv. 'KY­57, Datura stramonium were locally infected with the virus. The virus produced mosaic symptom on inoculated leaves of N. tabacum cv. 'Samson'. However, Chenopodium quinoa, Glycine max, Raphanus sativus, Cucumis sativus, Cucurbita moschata, Brassica rape and Lycopersion esculentum did not show any symptoms. TMV particles were revealed as a stiff rod shape by transmission electron microscopic(TEM) and measured as 300 nm in length with 18 nm in diameter. Total RNA was extracted from showing symptom loaves infected with TMV and the reverse transcription­polymerase chain reaction (RT­PCR) obtained 531 bp DNA product of RNA with specific primer used. The capsid protein of TMV­RE showed higher amino acid sequence homology(97.7%) with TMV­To than with TMV­P(72.2%). The capsid protein of TMV­152 showed same amino acid sequence homology with TMV­F. The result of comparison of nucleotides sequence homology between TMV­RE strain and other TMV strain showed 94% homology with others except TMV­P(67.3%) and TMV ­ C(68.6%).

Serological responses and protection levels in chickens administered with Newcastle disease vaccines

  • Geumji Seung;Jiye Kim;Hyobi Kim;Ji-Yeon Kim;Yang-Ho Jang;Yeon-Hee Kim;Moon Her;Seong-Joon Yi;Keun-Woo Lee;Il Jang;Young Ju Lee
    • Korean Journal of Veterinary Research
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    • v.62 no.4
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    • pp.29.1-29.7
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    • 2022
  • Vaccination against Newcastle disease (ND) is the most effective means of controlling the disease, and these vaccines are commercialized only after their safety and effectiveness have been verified through tests that comply with Korean Standards of National Lot Release for Veterinary Biologics. This study investigated whether a relatively convenient and safe serological test can be used in place of the challenge test using highly virulent ND virus. Hemagglutination inhibition (HI) assay and enzyme-linked immunosorbent assay (ELISA) were considered positive of log2 2 or more and cutoff value of 200 or more, respectively, in both live and inactivated vaccines. However, when the antibody levels of the live and inactivated vaccines induced using the Ulster 2C, KBNP-C4152R2L, and K148/08 strains were compared, the antibody titers for inactivated vaccines were significantly higher than those for live vaccines in both the HI assay and ELISA. A strong positive correlation was observed between HI and ELISA antibody titers. The live vaccines corresponded to a survival rates of ≥ 80% and the inactivated vaccines corresponded to 100% survival rates. This study confirmed that standard efficacy tests can serve as serological tests, and can replace the challenge test and that the vaccine approval process can be improved.

Effects of Sipyukmiyukieum on DNA Synthesis, cAMP Synthesis and MHC-class II Expression of FRTL-5 Thyroid Cells (십육미류기음(十六味流氣飮)이 FRTL-5 갑상선 세포의 DNA와 cAMP의 합성 및 MHC-class II의 발현에 미치는 영향)

  • Kim, Byoung-Woo;Yi, Jae-Eun
    • The Journal of Internal Korean Medicine
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    • v.26 no.2
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    • pp.398-408
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    • 2005
  • Objective: Graves' disease encompasses hyperthyroidism and diffused goiter associated with auto-antibodies to the thyroid stimulating hormone(TSH) receptors. In clinical environment, treatments of Graves' disease have many side effects such as recurrence and hypothyroidism. We've studied the effects of Sipyukmiyukieum on DNA synthesis, cAMP synthesis, and MHC-class II expression of FRTL-5 thyroid cells were studied. Methods: DNA synthesis was investigated by using BrdU staining and cAMP synthesis by ELISA kit, and expression of $interferon-{\gamma}$ activated MHC class II by Flow cytometer. Results: After introduction of Sipyukmiyukieum, significant inhibition of DNA synthesis. cAMP synthesis, and expression of $interferon-{\gamma}$ activated MHC class II of FRTL-5 thyroid cells was observed. Conclusions: Judging from these results, Sipyukmiyukieum has potential as a potent herbal treatment for inhibiting the enlargement of goiter, synthesis of abnormal thyroidal hormones, and autoimmuine responses of Graves' disease.

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Immunomodulatory Effects of Callophyllis japonica Ethanol Extract on Dendritic Cells (수지상세포에 대한 Callophyllis japonica 추출물의 면역조절효과)

  • Kim, Mi-Hyoung;Joo, Hong-Gu
    • IMMUNE NETWORK
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    • v.7 no.2
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    • pp.95-100
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    • 2007
  • Background: A red seaweed, Callophyllis japonica has been traditionally eaten in the oriental area. In a recent study, it has been demonstrated that the ethanol extract of C. japonica have antioxidant activity. However, there are few studies about the effects of C. japonica on the function of immune cells. We investigated the immunomodulatory effects of C. japonica on the function of dendritic cells, the potent antigen-presenting cells. Methods: Bone marrow-derived dendritic cells (DCs) were used and the viability was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and trypan blue exclusion test. Cytokine and nitric oxide (NO) levels were determined by using ELISA and Griess reagent, respectively. The expression levels of DC surface markers were measured by flow cytometric analysis. Results: C. japonica ethanol extract did not significantly affect the DCs viability and the IL-12 production from DCs, irrespective of the presence of lipopolysaccharide (LPS). In addition, it did not significantly change the expression of DC surface markers. However, C. japonica ethanol extract significantly inhibited the LPS-induced NO production and also increased the proliferation of allogeneic lymphocytes activated by DCs. Conclusion: Our data suggests that C. japonica ethanol extract enhances the proliferation of allogeneic lymphocytes activated by DCs which is associated with inhibition of NO production from DCs induced by LPS.

Generation of a monoclonal anti-human $\beta$2-adrenergic receptor antibody using GST-$\beta$-adrenergic receptor C-terminal fusion proteins expressed in E.Coli.

  • Kang, Suk-Jo;Shin, Chan-Young;Park, Kyu-Hwan;Ko, Kwang-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.95-95
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    • 1997
  • Among the various receptor molecules discovered so far the ${\beta}$2-adrenergic receptors have been regarded as excellent model systems for the so called 7 transmembrane helix receptor and have been the focus of extensive studies. For the analysis of receptor structure and function a monoclonal antibody plays a crucial role, thus providing useful tools for the study of receptor. However, because of the minute quantity of receptor molecules which could be obtained from natural sources, the generation of specific monoclonal antibody against receptor molecules from the purified receptors has been regarded as virtually impractical in consideration of cost and experimental times. The purpose of the present study was to generate and characterize a monoclonal antibody against human ${\beta}$2-adrenergic receptor. For the production of antibody, C-terminal regions of the human ${\beta}$2-adrenergic receptor was produced as a fusion protein with Glutathion S-transferase (GST) in E. Coli. The expression of the fusion protein was identified by SDS-PAGE and Western blot using monoclonal anti-GST antibody. The fusion protein was purified to an apparent homogeniety by affinity chromatography with Glutathion Sepharose CL-4B and used as an antigen for the immunization of BALB/C mice. The Production of monoclonal antibody was achieved by fusion of the immunized spleen cells and SP/2-0 myeloma cells. Positive hybridomas were screened by ELISA and were cloned by two consecutive rounds of limiting dilution. The monoclonal antibody produced in this study (mAb${\beta}$C02) was IgM type and purified by immunoaffinity chromatography using anti-mouse IgM agarose as an affinity matrix. MAb${\beta}$C02 showed strong and specific immunoreactivity against both the fusion protein and human ${\beta}$2-adrenergic receptor in ELISA and Western blot. The molecular weight of immunoreactive band was 64 kDa and exactly coincided with the previously reported molecular weight of ${\beta}$2-adrenergic recepters. The results of the present study suggest that mAb${\beta}$C02 may be used for the study of receptor function and regulation in normal or nonphysiological status.

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The Effect of Squalene on Inflammation Factors Induced by Candida Albicans in Vivo Studies (생체 내 상에서 Candida albicans로 유도한 염증인자에 미치는 스쿠알렌의 효과)

  • Lee, Jun-Haeng
    • Journal of radiological science and technology
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    • v.39 no.3
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    • pp.443-449
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    • 2016
  • In the present study, whether squalene treatment relives inflammatory reactions induced by Candida albicans was checked. The experiment was conducted in vivo using seven experimental animals (ICR mice) per experimental group. Among C. albicans-induced inflammatory factors, TNF-${\alpha}$, IL-6, and NO were observed using the ELISA kits method. Through the experiment, the following conclusions were obtained. 1. In the group infected with C. albicans, it could be identified that squalene treatment was inducing NO generation in renal tissues both on the 1st and 3rd days (p < 0.05). 2. In the group pre-treated(intraperitoneal administration) with SQ (80ml/kg) once per day for seven days and infected with C. albicans, it could be identified that squalene treatment was inducing TNF-${\alpha}$ generation in renal tissues only on the 3rd day(p < 0.05). 3. In the group pre-treated(intraperitoneal administration) with SQ (80ml/kg) once per day for seven days and infected with C. albicans, it could be identified that squalene treatment was inducing IL-6 generation in renal tissues only on the 3rd day(p < 0.05). In conclusion, it could be seen that for squalene to suppress C. albicans-induced inflammatory factors, preemptively supplying SQ should be effective. Therefore, effects for recovery from C. albicans-induced immunodepression can be expected from SQ treatment.

Anti-inflammatory effect of chloroform fraction of Coptidis rhizoma on the production of inflammatory mediators from LPS-stimulated BV2 microglial cells (황련 클로로포름 분획물의 뇌신경소교세포로부터 염증매개물질 생성억제 효능 연구)

  • Park, Yong-Ki;Lee, Kyuong-Yeol
    • The Korea Journal of Herbology
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    • v.22 no.4
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    • pp.109-116
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    • 2007
  • Objectives : In the present study, we investigated anti-inflammatory effects of chloroform fraction of Coptidis rhizoma (CR-C) on the production of inflammatory mediators such as nitric oxide (NO) and proinflammatory cytokines, tumor necrosis factor-alpha (TNF-${\alpha}$) and interleukin-1beta (IL-1${\beta}$) in LPS-stimulated BV2 microglial cells. Methods : Copriditis rhizoma was extracted with 80% methanol, and then extracted with chloroform. BV2 cells were pre-treated with CR-C, and stimulated with LPS. The cytotoxicity was determined by MTT assay. The production of NO and cytokines was measured by Griess assay and ELISA. The mRNA expression of inducible nirtic oxide synthase (iNOS) and cytokines were determined by RT-PCR. Results : CR-C significantly inhibited the production of NO. TNF-${\alpha}$ and IL-1${\beta}$ in a dose-dependent manner in LPS-stimulated BV2 cells. In addition, CR-C suppressed the mRNA expressions of iNOS and inflammatory cytokines induced by LPS stimulation. These results indicate that CR-C was involved in anti-inflammatory effects in activated microglia. Conclusion : The present study suggests that chloroform extract of Coptidis rhizoma can be useful as a potential anti-inflammatory agent for treatment of various neurodegenerative diseases.

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Time gap between oocyst shedding and antibody responses in mice infected with Cryptosporidium parvum

  • Yu, Jae-Ran;Lee, Soo-Ung
    • Parasites, Hosts and Diseases
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    • v.45 no.3
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    • pp.225-228
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    • 2007
  • We observed the time gap between oocyst shedding and antibody responses in mice (3-week-old C57BL/6J females) infected with Cryptosporidium parvum. Oocyst shedding was verified by modified acid-fast staining. The individually collected mouse sera were assessed for C. parvum IgM and IgG antibodies by enzyme-linked immunosorbent assay from 5 to 25 weeks after infection. The results showed that C. parvum oocysts were shed from day 5 to 51 post-infection (PI). The IgM antibody titers to C. parvum peaked at week 5 PI, whereas the IgG antibody titers achieved maximum levels at week 25 PI. The results revealed that IgM responses to C. parvum infection occurred during the early stage of infection and overlapped with the oocyst shedding period, whereas IgG responses occurred during the late stage and was not correlated with oocyst shedding. Hence, IgM antibody detection may prove helpful for the diagnosis of acute cryptosporidiosis, and IgG antibody detection may prove effective for the detection of past infection and endemicity.

Characteristics of Tobacco Mosaic Virus Isolated from Wasabi (Eutrema wasabi) in Korea

  • Kim, Hyung-Moo;Lee, Kui-Jae
    • The Plant Pathology Journal
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    • v.15 no.4
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    • pp.247-250
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    • 1999
  • Wasabies showing mosaic symptoms were collected and extracted for virus purification. Tobacco mosaic virus (TMV) was identified as causal agent by electron microscopy and nucleic acid and coat protein analyses. TMV strains were determined by enzyme-linked immunosorbent assay (ELISA). TMV was identified as W and C strain in wasabi. The results of host reaction indicated that this virus induced local lesions on Nicotiana tabacum cv. Bright Yellow and N. glutinosa, leaf spots on Chenopodium amaranticolor and mosaic symptoms on wasabi. Rot shape virus particles were observed and was about 300 nm in length. About 6.5 kb single RNA molecule was observed from extracted viral RNA sample and 26 KDa coat protein was detected in denatured acrylamide gel. Infection ratio of TMV was 8% for the first cultivation year, but was 22% for the second year when TMV-W antiserum was used. The results of this experiment showed that infection ratios of both TMV-W and TMV-C strains were higher compared to that of TMV-P strain.

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The Effects of Phragmitis Rhizoma Herbal-acupuncture Solution on Inflammation in Human Mast Cells and Human Alveolar Epithelial Cell Lines - Phragmitis Rhizoma's Effects -

  • Kim, Byung-Soo
    • The Journal of Korean Medicine
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    • v.35 no.4
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    • pp.1-9
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    • 2014
  • Objectives: This study was designed to find the effect of Phragmitis Rhizoma (PR) herbal-acupuncture solution on the inflammatory cytokine and chemokine secretion in human mast cell (HMC) and human alveolar epithelial cell 549 (A549) lines. Methods: Histamine levels in HMC after PR herbal-acupuncture solution treatment were measured with ELISA. Other cytokines and chemokines levels such as interleukin 8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and chemokine (C-C motif) ligand 5 (Ccl5, RANTES) in A549 were measured with flow cytometry CBA system. Results: In the PR herbal-acupuncture solution treatment group, the expression of histamine, IL-8, MPC-1, Ccl5, and RANTES decreased significantly. Conclusions: The results support that PR herbal-acupuncture solution had a suppressive effect on cytokine-induced inflammation.