• Title/Summary/Keyword: IgA

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The Reactivity of Antiserum Raised against Native Glucose-6-phosphate Dehydrogenase with Denatured Glucose-6-phosphate Dehydrogenase in Competitive ELISA

  • Kim, Moon-Hee
    • BMB Reports
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    • v.31 no.5
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    • pp.519-523
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    • 1998
  • We have previously reported that anti-glucose-6-phosphate dehydrogenase (G6PD) serum raised against native G6PD (nG6PD) enzyme recognized nG6PD antigen poorly in competitive enzyme-linked immunosorbent assay (ELISA) (Kim, 1997). In the present study, we investigated whether anti-G6PD serum raised against nG6PD can react with denatured G6PD effectively in competitive ELISA. We used partially active G6PD (paG6PD) by repeated freeze-thawing or SDS-denatured G6PD (SDS-G6PD) as both immobilized and soluble antigens, and anti-G6PD serum raised against nG6PD for competitive ELISA. The polystyrene cuvettes coated with either paG6PD or SDS-G6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of paG6PD or SDS-G6PD as competitors, followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA with paG6PD or SDS-G6PD antigen exhibited the sigmoidal dose-response curve characteristic of competition immunoassays. Furthermore, Triton-denatured G6PD (Triton-G6PD) was used in competitive ELISA. The paG6PD, SDS-G6PD, or Triton-G6PD used as competitors increased the inhibition of antibody binding to immobilized either of nG6PD or denatured G6PD compared with nG6PD competitor. The inhibition by denatured G6PD competitors was more pronounced at high competitor concentrations than at low counterparts. We conclude that anti-G6PD serum raised against nG6PD can effectively react with denatured G6PD in competitive ELISA and that our anti-G6PD serum recognizes denatured enzymes better than active enzymes.

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Anti-asthmatic Effects of Samjajihwang-tang in OVA-induced Mice (삼자지황탕(三子地黃湯)의 생쥐 모델에 대한 항천식 효과)

  • Kim, Woon-Kil;Park, Yang-Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.2
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    • pp.343-350
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    • 2009
  • This study aimed to evaluate the anti-asthmatic effects of Samjajihwang-tang (SJT) using OVA-induced asthmatic mice model. Asthmatic mice model was conducted by repeated challenge of OVA using C57BL/6 mice. Each group was treated with distilled water, SJT (400 mg/kg and 200 mg/kg) extract or cyclosporin A (10 mg/kg) for the later 8 weeks, Penh (plethysmography and enhanced pause), immune cells subpopulation, eotaxin, IL-5, TNF-${\alpha}$, Anti-OVA-lgE in BALF (bronchoalveolar lavage), and lung tissue was analyzed, No cytotoxicity of SJT was shown on hFCs (human fibroblast cells). Administration of SJT significantly decreased Penh levels comparing to control group. SJT treatment significantly ameliorated the increase of total cells number and eosinophil including of immune cell subpopulation of $CD3^+/CD69^+$, $CCR3^+$, $B220^+/CD22^+$, $B220^+/CD45^+$, and $B220^+/lgE^+$ cells in BALF comparing to control group. Eotaxin, IL-5, TNF-${\alpha}$, and Anti-OVA-lgE level in BALF were significantly decreased by SJT treatment too. Histopathological finding verified the improvement of infiltration of inflammatory cells and collagen tissue in the SJT groups comparing to control group. These results strongly suggest that SJT would be a effective candidate for herbal-originated anti-asthmatic drug. However, this drug should be further studied for characterization of the accurate action and underlying mechanisms using variant disease model in the future.

A dominant hyperrectangle generation technique of classification using IG partitioning (정보이득 분할을 이용한 분류기법의 지배적 초월평면 생성기법)

  • Lee, Hyeong-Il
    • Journal of the Korea Society of Computer and Information
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    • v.19 no.1
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    • pp.149-156
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    • 2014
  • NGE(Nested Generalized Exemplar Method) can increase the performance of the noisy data at the same time, can reduce the size of the model. It is the optimal distance-based classification method using a matching rule. NGE cross or overlap hyperrectangles generated in the learning has been noted to inhibit the factors. In this paper, We propose the DHGen(Dominant Hyperrectangle Generation) algorithm which avoids the overlapping and the crossing between hyperrectangles, uses interval weights for mixed hyperrectangles to be splited based on the mutual information. The DHGen improves the classification performance and reduces the number of hyperrectangles by processing the training set in an incremental manner. The proposed DHGen has been successfully shown to exhibit comparable classification performance to k-NN and better result than EACH system which implements the NGE theory using benchmark data sets from UCI Machine Learning Repository.

Study on the Immune Modulatory Activity of Seokjahaeki-tang Using Atopic Dermatitis Animal Models (아토피피부염 동물 병태 모델에서 석자해기탕(石紫解肌湯)의 면역조절작용에 관한 연구)

  • Lee, Byung-Woong;Gim, Seon-Bin;Song, Hyang-Hee;Ji, Joong-Gu;Bak, Ji-Won;Kim, Dong-Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.4
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    • pp.446-454
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    • 2012
  • In order to evaluate the efficacy of SHT against atopic dermatitis (AD), various immune related cytokines as well as histological comparison were performed in animal models, and the results are described. Clinical skin index of the SHT treated group decreased significantly in weeks 11 and 13, compared to the control group. Also, CD4+ immune cell ratio in the dorsal skin was significantly decreased to 69%, and both epidermal and dermal skin thickness was decreased. Serum IL-4, IL-5, IL-6, IL-13, and TNF-${\alpha}$, which are all important markers of inflammation, were decreased to 64%, 44%, 87%, 48%, and 45%, respectively. The expression of histamine, a chemical transmitter increasingly released during the progression of inflammation, was significantly decreased to 47%. The production of IgE immunoglobulin was significantly decreased to 16% compared to the control group. In conclusion, SHT pacifies the activation of T cells, leading to suppression of both Th2 cytokine overexpression and infiltration of immune cells into skin. As a result, relative thinning of both epidermis and dermis were observed. With the results obtained from in vitro studies, the immune modulatory effect of SHT in AD animal models was experimentally demonstrated. This study should provide solid information to construct EBM and for clinical practice.

Studies on avian infectious bronchitis: II. Standardization of an indirect enzyme-linked immunosorbent assay (ELISA) for antibody measurement (닭 전염성 기관지염에 관한 연구: II. 간접 enzyme-linked immunosorbent assay(ELISA)에 의한 항체가 측정)

  • Chang, Chong-ho;Kim, Sun-joong
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.503-515
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    • 1989
  • Critical parameters affecting sensitivity and specificity of an enzyme-linked immunosorbent assay(ELISA) for detection of antibodies to avian infections bronchitis virus(IBV) were standardized. By adopting the optimized conditions an equation calculating ELISA antibody titers from the observations at single serum dilution was formulated. The purified antigen of IBV Mass-41 strain was dispensed into polystyrene microplate wells at a concentration of 300ng per well($100{\mu}l$) and the plates were coated by completey drying at $37^{\circ}C$. Diluted chicken serum and horseradish peroxidase conjugated goat anti-chicken IgG were added in order in $100{\mu}l$ volumes per well and allowed to react for 30 minutes each at room temperature. Just before use and after each reaction the plates were washed three times with distilled water. Finally o-phenylenediamine solution was added as an enzyme substrate. After incubation for another 15 minutes at room temperature absorbances were read at 492nm. Hyperimmune serum against Mass-41 strain was used as internal reference positive(IRP) serum. After repeated titration of IRP and negative serum, a constant titer of IRP was determined. Serum titrations were carried out for various sample sera together with IRP and negative sera and the observed titers of sample sera were corrected by reflecting the ratio between observed and constant titers of IRP serum. These corrected titers of the sample sera were plotted against sample/positive(S/P) OD ratios. All the OD's measured in the serum titrations were also corrected by substracting negative serum OD. The following equation was formulated from the above data; $Log_{10}$ ELISA titer=$5.568({\log}_{10}S/P)+4.161$ Thus it was possible to calculate ELISA titer by measuring absorbance at 1/400 single serum dilution. Titer measured by cross ELISA tests employing Mass-41 strain and three local IBV isolates were similar. These results suggest that the ELISA tests standardized in this study can be used for evaluating not only vaccinal immunity but also for infection status against fields IBV's.

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Efficacy of orally administered ginseng stem and leaf in chickens (인삼 지상부를 첨가한 사료 급여가 닭에서 나타내는 효과)

  • Park, Soyeon;Lee, Kwang-Yeal;Cho, Youngjae;Park, Bokyoung;Kim, Kiju;Lee, Na-Rae;Kim, Dong-Gun;Kim, Young-Hee;Hahn, Tae-Wook
    • Korean Journal of Veterinary Research
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    • v.55 no.1
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    • pp.1-7
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    • 2015
  • Ginseng has been widely used in Korea as a natural medicine due to its saponin contents. Although the total amount of ginseng stem and leaf saponins (GSLS) is 4~5 times higher than that of saponin in the root, the root is mainly used. This is due to two reasons: nervous system-stimulant activity of GSLS and pesticide residues in GSLS. In this study, residual agricultural pesticides were removed from GSLS using two types of bacterial treatments. Two GSLS treatment groups of chickens (GSLS-1 and GSLS-2) were established. The chickens were fed 0.4% GSLS-1 or GSLS-2 mixed with crop. We then evaluated the effects of GSLS on bodyweight and several immune parameters. At the end of the experiments, chickens fed GSLS-1 and red ginseng saponin had significantly higher growth rates (16.6% and 8.0%, respectively) compared to the vaccine control group treated with Noblis Salenvac-T. The group fed GSLS-1 also had the highest IgG titer that was significantly different at the end of experiments compared to the other groups. These findings imply that GSLS-1 is a good candidate feed additive for the chicken industry.

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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Effect of Polygonum multipolarum extract on the allergic reaction of NC/Nga mice causing atopic dermatitis (하수오 추출물(Polygonum multiflorum)이 아토피 피부염 유발 NC/Nga 생쥐의 알레르기 반응에 미치는 영향)

  • Moon, Ji-Sun;Choi, Eun-Young
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.1
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    • pp.66-75
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    • 2020
  • Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. and wished to examine closely effect that Polygonum multiflorum isolated PM-E and PM-70M orally adminstration used to atopy dermatitis disease patient get in atopy eruption control experimentally. Atopic dermatitis is a chronically relapsing inflammatory skin disease. Animal models induced by relevant allergens play a very important role in the elucidation of the disease. This study was investigated the anti-allergic effect of PM-E and PM-70M on BMAC induced atopic dermatitis in NC/Nga mice. We summerized as the follow. PM-E and PM-70M significantly reduced the skin number of total cell number, CD4+ and CD11b+/Gr-1 cell compared with positive control and decreased the invasion of CD4+ cell in dorsal skin tissue compared with positive control group by using immunohistochemical staining and chemokine such as eotaxin and CCR3 compared with positive control group. PM-E and PM-70M markedly suppressed invasion and edema of leukocytes and mast cell in dorsal skin. Taken together, these findings suggested that PM-E and PM-70M has an anti-allergic activity and this might be useful for the clinical application to treat allergic diseases such as atopic dermatitis.

Vitamin D serum levels in children with allergic and vasomotor rhinitis

  • Lee, Seung Jin;Kang, Bong Hwa;Choi, Bong Seok
    • Clinical and Experimental Pediatrics
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    • v.58 no.9
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    • pp.325-329
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    • 2015
  • Purpose: In addition to regulating calcium and phosphorus homeostasis and bone metabolism, vitamin D is known as an immune modulator. Recently, there has been increased worldwide interest in the association between low levels of vitamin D and allergic diseases. The purpose of this study was to assess the relationship between serum vitamin D levels and allergic/vasomotor rhinitis (AR/VR) in children. Methods: This study included 164 patients. The sample included 59 patients with AR, 42 patients with VR, and 63 controls. Their ages ranged from 0 to 16 years. We examined the levels of 25-hydroxyvitamin D, Immunoglobulin E, specific IgE, and eosinophil cationic protein; peripheral blood eosinophil count; and the results of a skin prick test. Results: Serum 25-hydroxyvitamin D levels were $19.0{\pm}8.5ng/mL$ in the AR group, $25.5{\pm}10.9ng/mL$ in the VR group, and $26.9{\pm}10.7ng/mL$ in the control group. After adjustment for body mass index and season at the time of blood sampling, vitamin D levels in the AR group were lower than those of the VR group (P=0.003) and control group (P<0.001). Vitamin D levels were inversely correlated with Immunoglobulin E levels (r=-0.317, P<0.001). AR patients with food allergy or atopic dermatitis did not have lower levels of 25-hydroxyvitamin D than AR patients without these diseases. Conclusion: This study demonstrates a possible relationship between vitamin D levels and allergic rhinitis in Korean children.

Oral food challenges in children

  • Yum, Hye-Yung;Yang, Hyeon-Jong;Kim, Kyung-Won;Song, Tae-Won;Kim, Woo-Kyung;Kim, Jung-Hee;Ahn, Kang-Mo;Kim, Hyun-Hee;Lee, Soo-Young;Pyun, Bok-Yang
    • Clinical and Experimental Pediatrics
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    • v.54 no.1
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    • pp.6-10
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    • 2011
  • Many patients assume that allergic reactions against foods are responsible for triggering or worsening their allergic symptoms. Therefore, it is important to identify patients who would benefit from an elimination diet, while avoiding unnecessary dietary restrictions. The diagnosis of food allergy depends on the thorough review of the patients's medical history, results of supplemented trials of dietary elimination, and in vivo and in vitro tests for measuring specific IgE levels. However, in some cases the reliability of such procedures is suboptimal. Oral food challenges are procedures employed for making an accurate diagnosis of immediate and occasionally delayed adverse reactions to foods. The timing and type of the challenge, preparation of patients, foods to be tested, and dosing schedule should be determined on the basis of the patient's history, age, and experience. Although double-blind, placebo-controlled food challenges(DBPCFC) are used to establish definitively if a food is the cause of adverse reactions, they are time-consuming, expensive and troublesome for physician and patients. In practice, An open challenge controlled by trained personnel is sufficient especially in infants and young children. The interpretation of the results and follow-up after a challenge are also important. Since theses challenges are relatively safe and informative, controlled oral food challenges could become the measure of choice in children.