• Title/Summary/Keyword: isolation performance

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Immunogenicity of Synthetic Peptide Specific for Major Immunogenic Determinat of Hepatitis B Surface Antigen (B형간염(型肝炎) 표면항원(表面抗原)의 주면역원(主免疫原) 결정기(決定基)에 특이(特異)한 합성(合成) Peptide의 면역원성(免疫原性)에 관한 연구(硏究))

  • Shin, Kwang-soon;Han, Su-nam
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.7-17
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    • 1985
  • Many investigators have been pursuing various attempts so far to produce hepatitis B surface antigen(HBsAg) vaccines using the techniques such as isolation from plasma of chronic HBsAg carrier, recombinant DNA technique or preparation of synthetic peptides specific for immunogenic determinants. Hepatitis B virus can not grow on any cell lines by the tissue culture technique at the present time. The plasma of chronic HBsAg carrier is expensive and its source is limited. The HBsAg from the recombinant DNA technique gave still very low yield. Another approach, therefore, has been initiated to develop a synthetic hepatitis B virus vaccine. The possible use of several distinct synthetic vaccines in prophylaxis can be facilitated by availability of full synthetic immunogens. Peptides synthesized for potential application as antiviral vaccines have been mostly tested in the form of conjugates with carrier proteins, although the free synthetic peptide can be immunogenic. To understand basic knowledges on the antigenicity and immunogenicity of a synthetic peptide specific for major immunogenic determinant of HBsAg, a nonapeptide, $H_2N^{139}Cys-Thr-Lys-Pro-Thr-Asp-Gly-^{146}Asn-Aba$ COOH, which corresponds to HBsAg amino acid residues 139 to 147, was synthesized by the Merrifield's solid-phase method with a slight modification. The antigenicity and immunogenicity of this specific synthetic peptide were examined comparing with purified plasma-derived natural HBsAg. The results obtained are as follows; 1. The peptide synthesized showed the identical amino acid composition to the theoretical value. The degree of purification and molecular weight were acertained by methods of high performance liquid chromatography and mass spectrometry. 2. Using m-maleimidobenzoyl-N-hydroxysuccinimide ester as a conjugating agent, the synthetic peptide was conjugated to rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin. Their conjugation yields were 8.3, 9.5, 15.8, 13.5, and 11.2%, respectively. 3. The natural HBsAg was purified from plasma of chronic HBsAg carrier. By the electron microscopic observation of the purified natural HBsAg preparation, no Dane particles were observed and the preparation showed negative DNA polymerase activity. 4. Antigenicity of the synthetic peptide and the plasma-derived natural HBsAg was determined by competition radioimmunoassay using $^{125}I$-natural HBsAg. Their 50% inhibitions appeared as $90{\mu}g/ml$ and $0.12{\mu}g/ml$ for the synthetic peptide and the natural HBsAg, respectively. This indicates that the former was about 750-fold less antigenic than the latter. 5. Immunogenicity of the synthetic peptide was determined by administering the peptide-carrier conjugates into rabbits with and without Freund's complete adjuvant. Regardless the carrier proteins and adjuvant, positive immune responses to the synthetic peptide were observed. The higher antibody titers, however, were shown in the groups administered with Freund's complete adjuvant. 6. Immunizing dose 50% in mice of the various peptide-carrier conjugates was 5.47, 6.00, 65.16, 31.25 and $13.03{\mu}g/dose$ for rabbit albumin and ${\gamma}$-globulin, tetanus and diphtheria toxoids, and keyhole limpet hemocyanin, respectively, while the natural HBsAg showed $0.65{\mu}g/dose$. 7. It was postulated that homologous proteins prefer to heterologous ones as the carriers.

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Nutrition and Psychosocial Factors were associated with Possible Sarcopenia in the Rural Elderly (농촌지역 주민의 영양결핍 및 사회심리적 요인과 근감소증 가능군과의 관련성)

  • Kim, Bokyoung;Lee, Gyeong-Ye;Seo, Ae-Rim;Kim, Mi-Ji;Seo, Sung-Hyo;Park, Ki-Soo
    • Journal of agricultural medicine and community health
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    • v.47 no.2
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    • pp.90-98
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    • 2022
  • Objective: This study aimed to provide basic data for preventing and managing sarcopenia by identifying the relationship between sarcopenia, malnutrition, and psychosocial factors among the elderly in the community. Methods: The study included 1,019 subjects aged 60 and over. "Possible sarcopenia" was defined by low handgrip strength with or without reduced physical performance. Nutrition was evaluated according to the mini nutrition assessment (MNA), and the psychosocial factors examined were self-efficacy, social isolation, fear of falling, and social capital (trust and participation). A logistic regression analysis was also performed on the relationship between risk of malnutrition, psychosocial factors, and sarcopenia. Results: MNA was significantly associated with social participation (OR = 1.747, p <0.001), fear of falling (OR = 2.905, p <0.001), and self-efficacy (high/low, OR = 0.654, p = 0.011). In model 3, which included both MNA and psychosocial factors, sarcopenia was significantly associated with MNA (OR = 2.529, p <0.001) and fear of falling (OR = 1.544, p = 0.045). Compared with the low self-efficacy group, the high group (OR = 0.589, p = 0.009) was significant. The factors related to possible sarcopenia include risk of malnutrition, fear of falls, and low self-efficacy. Conclusion: It will be necessary to improve self-efficacy so that individuals feel they can do activities of daily living themselves and to reduce their fear of falling through muscle strength and balance exercises. Finally, it is also necessary to increase regular participation in community social activities.

Development and Evaluation of Multiplex PCR for the Detection of Carbapenemase-Producing Enterobacteriaceae (카바페넴분해효소 생성 장내세균 검출을 위한 Multiplex PCR의 개발 및 평가)

  • Kim, Si Hyun;Bae, Il Kwon;Kim, Na Young;Song, Sae Am;Kim, Sunjoo;Jeong, Joseph;Shin, Jeong Hwan
    • Annals of Clinical Microbiology
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    • v.22 no.1
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    • pp.9-13
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    • 2019
  • Background: The isolation of carbapenemase-producing Enterobacteriaceae (CPE) has become increasingly common. Continuous surveillance for these organisms is essential because their infections are closely related to outbreaks of illness and are associated with high mortality rates. The aim of this study was to develop and evaluate multiplex PCR as a means of detecting several important CPE genes simultaneously. Methods: We aimed to develop a multiplex PCR that could detect seven CPE genes simultaneously. The multiplex PCR was composed of seven primer sets for the detection of KPC, IMP, VIM, NDM-1, GES, OXA-23, and OXA-48. We designed different PCR product sizes of at least 100 bp. We evaluated the performance of this new test using 69 CPE-positive clinical isolates. Also, we confirmed the specificity to rule out false-positive reactions by using 71 carbapenem-susceptible clinical strains. Results: A total of 69 CPE clinical isolates showed positive results and were correctly identified as KPC (N=14), IMP (N=13), OXA-23 (N=12), OXA-48 (N=11), VIM (N=9), GES (N=5), and NDM (N=5) by the multiplex PCR. All 71 carbapenem-susceptible clinical isolates, including Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, showed negative results. Conclusion: This multiplex PCR can detect seven CPE genes at a time and will be useful in clinical laboratories.