• Title/Summary/Keyword: Immunization technique

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Detection of Abnormally High Amygdalin Content in Food by an Enzyme Immunoassay

  • Cho, A-Yeon;Yi, Kye Sook;Rhim, Jung-Hyo;Kim, Kyu-Il;Park, Jae-Young;Keum, Eun-Hee;Chung, Junho;Oh, Sangsuk
    • Molecules and Cells
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    • v.21 no.2
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    • pp.308-313
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    • 2006
  • Amygdalin is a cyanogenic glycoside compound which is commonly found in the pits of many fruits and raw nuts. Although amygdalin itself is not toxic, it can release cyanide (CN) after hydrolysis when the pits and nuts are crushed, moistened and incubated, possibly within the gastrointestinal tract. CN reversibly inhibits cellular oxidizing enzymes and cyanide poisoning generates a range of clinical symptoms. As some pits and nuts may contain unusually high levels of amygdalin such that there is a sufficient amount to induce critical CN poisoning in humans, the detection of abnormal content of amygdalin in those pits and nuts can be a life-saving measure. Although there are various methods to detect amygdalin in food extracts, an enzyme immunoassay has not been developed for this purpose. In this study we immunized New Zealand White rabbits with an amygdalin-KLH (keyhole limpet hemocyanin) conjugate and succeeded in raising anti-sera reactive to amygdalin, proving that amygdalin can behave as a hapten in rabbits. Using this polyclonal antibody, we developed a competition enzyme immunoassay for determination of amygdalin concentration in aqueous solutions. This technique was able to effectively detect abnormally high amygdalin content in various seeds and nuts. In conclusion, we proved that enzyme immunoassay can be used to determine the amount of amygdalin in food extracts, which will allow automated analysis with high throughput.

Studies on the Development and Utilization of Polyclonal Antibodies Against Swine Adipocyte Plasma Membrane Proteins (돼지 지방세포 원형질막 단백질에 대한 다클론항체의 생산 및 이용에 관한 연구)

  • Baek, K.H.;Kwak, T.H.;Oh, Y.S.;Choi, C.W.;Jung, K.K.;Choi, Chang-Bon
    • Journal of Animal Science and Technology
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    • v.47 no.1
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    • pp.19-28
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    • 2005
  • The objectives of the current study were to develop polyclonal antibodies in sheep against adipocyte plasma membrane(APM) proteins isolated from swine, to investigate tissue specificity, and to determine cytotoxic effects of antiserum on swine adipocytes. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver, and spleen were isolated using a 32% sucrose gradient. Adult male sheep was immunized three times at three week interval with the purified swine APM proteins. Antiserum was taken from immunized sheep at 10, 12, and 14 days after the third immunization. Antiserum expressed strong reactivity with APM proteins determined by enzyme-linked immunosorbent assay(ELISA), and the reactivity could be detected at dilutions in excess of 1 : 81,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver, or spleen. Tissue specificity of the antiserum was reconfirmed by Western immunoblotting using anti-sheep immunoglobulin G•alkalinephosphatase conjugate as a secondary antibody. The reactivity of antiserum to the external surface of fixed swine adipocytes was confmned by an immunohistochemical technique using anti-sheep immunoglobulin G-FITC. Confluent swine adipocytes in culture were lysed by antiserum treatment and cytosolie lactate dehydrogenase(LDH) was released as a dose-dependent patterns while adipocytes treated with normal sheep serum maintained their integrity and expressed low level of LDH. These results implicate that fat contents in the pigs can be reduced by immunological methods.

Evaluation on Effectiveness of Public Health Personnel Training (보건요원 교육효과 평가)

  • Hwang Keum-Bok;Jeon Mi-Soon;Kim Tae-Sook;Yang Byung-Guk;Jeong Eun-Kyung;Yang Sook-Ja;Kim Kwuy-Hyang
    • The Journal of Korean Academic Society of Nursing Education
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    • v.5 no.1
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    • pp.118-132
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    • 1999
  • To improve the effectiveness and efficiency of public health personnel training, we evaluated not only how appropriate the students felt the objectives, contents, methods and multimedia used in the train ing courses, but also how much the students accomplished the objectives and applied skill and knowledge to their own works. We selected 5 courses for the study : Tuberculosis control, Radiological technique, Public health information, Immunization, Mental health management courses used by Kirkpatrick's evaluation model. Reaction evaluation was carried out in final day by questionnaire. The results showed that all of them were very satisfied with educational input and curricula, learn Ing environment. Secondly, we measured the degree of learning achievement on pre and post training by questionaire of specific behavioral objectives. The degree of learning achievement was statistically higher just after training than pre training (paired t-test, p<0.01). Thirdly, evaluation of behavioral change to job was conducted to find out how much students applied skill and knowledge to their own job in 3 months after training by questionnaire. The results of behavioral change evaluation showed that 43.5% of the students who were performing job related with the training courses in 3 months after training applied the learned skill and knowledge to their own job quite well and 37.8% of them applied relatively well, therefore total 81.4% of them applied to their own job. And effectiveness of training for the above mentioned students showed that 41.9 % of them had improved or enforced their jobs after training, 35.5% of them had had no remarkable changes, and 15.7% had newly applied the learned skill and knowledge to their jobs. For evaluating the degree of usefulness of material predistribution in two weeks before training, we compared experimental groups with control groups. The results showed that general reactions are helpful but the degree of learning achievement is no discrepancy.

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$RpoB_{127-135}$ Peptide Derived from Mycobacterium tuberculosis is Processed and Presented to HLA-$A^*0201$ Restricted CD8+ T Cells via an Alternate HLA-I Processing Pathway

  • Cho, Jang-Eun;Cho, Sang-Nae;Cho, Sungae
    • Biomedical Science Letters
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    • v.20 no.4
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    • pp.250-255
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    • 2014
  • Mycobacterium tuberculosis (MTB) resides and replicates inside macrophages. In our previous report, we reported that CD8+ T cell-mediated immune responses specific for the peptide derived from MTB RNA polymerase beta-subunit ($RpoB_{127-135}$) could be induced in TB patients expressing HLA-$A^*0201$ subtype. In order to examine whether $RpoB_{127-135}$ specific CD8+ T cells can recognize MTB infected macrophages in vitro, CD8+ T cell lines specific for $RpoB_{127-135}$ peptide were generated from peripheral blood mononuclear cells (PBMCs) of healthy HLA-$A^*0201$ subjects by in vitro immunization technique. In this study, we observed $RpoB_{127-135}$ specific CD8+ T cells could recognize and destroy macrophages infected with MTB for 2 to 4 days. $RpoB_{127-135}$ specific CD8+ T cell immune response was inducible from PBMC of healthy subjects expressing HLA-$A^*0206$ subtype, one of HLA-A2 supertype members. Next, we investigated the HLA-I processing mechanism of $RpoB_{127-135}$ peptide in MTB infected macrophages. As a result, the presentation of the MTB derived epitope peptide, $RpoB_{127-135}$, to CD8+ T cells was not inhibited by the treatment with brefeldin-A (ER-Golgi transport inhibitor) or lactacystin (proteasome inhibitor), which blocks the classical HLA-I processing pathway. However, $RpoB_{127-135}$ specific CD8+ T cell activity was blocked either by the blocking agent for the endocytosis (cytochalasin D) or by the blocking antibody (W6/32) for HLA-I molecules. Therefore, the $RpoB_{127-135}$ peptide may be processed by accessing the alternate HLA-I processing pathway. Understanding the processing and presentation mechanisms of the MTB derived proteins will help to improve the efficacy of vaccines and the efficiency of therapeutic agents for TB.

Delphi Survey for COVID-19 Vaccination in Korean Children Between 5 and 11 Years Old (국내 5-11세 소아의 코로나19 백신 접종에 대한 델파이 연구)

  • Choe, Young June;Lee, Young Hwa;Choi, Jae Hong
    • Pediatric Infection and Vaccine
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    • v.29 no.1
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    • pp.37-45
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    • 2022
  • Purpose: During the coronavirus disease 2019 (COVID-19) pandemic, we conducted a Delphi survey that included the experts from the field of COVID-19 immunization in children aged 5-11 years. The aim was to organize collective expert opinions on COVID-19 vaccination in young children in the Republic of Korea, and so thus assist the vaccination policy. Methods: The panels included pediatric infectious disease specialists, preventive medicine experts, infectious disease physicians, and COVID-19 vaccine experts consulting the Ministry of Health and Welfare. The Delphi survey was conducted online using a questionnaire from February 14 to February 27, 2022. Results: The Delphi panels agreed that children were vulnerable to COVID-19, and the severity of illness was modest. Furthermore the panels reported that children with chronic illness were more susceptible to a worsening clinical course. There were generally positive opinions on the effectiveness of COVID-19 vaccination in children aged 5-11 years, and experts gathered a slightly positive opinion that the adverse events of pediatric COVID-19 were not numerous. The benefits of COVID-19 vaccination were evaluated at a level similar to the potential risks in children. Currently, the only approved mRNA platform vaccine in children seemed to be sustainable; however, the recombinant protein platform COVID-19 vaccines were evaluated as better options. Conclusions: Due to the surge of the Omicron variant and an increase in pediatric cases, the COVID-19 vaccination in young children may have to be considered. Panels had neutral opinions regarding the COVID-19 vaccination in children aged 5-11 years. Thus monitoring of the epidemiology and the data about the safety of COVID-19 vaccination should be continued.