• Title/Summary/Keyword: immunodiffusion

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Pulmonary Fungal Infection in Patients with Tuberculosis or Other Lung Diseases and Fungal Flora in Human Sputum Specimens (폐결핵 또는 기타 폐질환 환자에서의 진균감염과 객담으로부터 분리되는 임상적유의 및 무의진균에 관한 연구)

  • Kim, Sang-Jae;Hong, Young-Pyo;Kim, Sin-Ok;Lee, Ho-Won;Lee, Seok-Gi
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.1
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    • pp.15-22
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    • 1987
  • Pulmonary fungal infection has been investigated in patients with healed or active pulmonary tuberculosis or other lung diseases by demonstrating serum precipitating antibodies to the various fungal antigens and by isolating the related fungi from sputums or other clinical specimens. Out of 1,192 suspected patients, 405(34.0%) showed precipitin bands on immunodiffusion tests and the related fungi have been demonstrated in sputums or other specimens of 79.5% of them(327) whose specimens had been cultured. Three patients did not have precipitating antibodies to any fungal antigen, but the same fungus was repeatedly isolated from sputums of two patients for over one year of period and from open lung biopsy specimen in the other patient. Most commonly involved species in pulmonary infection were A. fumigatus(70.3%) and C. albicans (at least 23.8%), followed by A. flavus, P. boydii, A. nidulans, etc. Out of fungi isolated from individuals(459), who were apparently not infected with fungi, molds were 66.0% and the rest, yeasts. Most commonly encountered molds were aspergilli(31.7%), followed by penicilli(16.3%), Cladosporium spp.(2.8%), Fusarium spp.(2.2%), etc. C. albicans(16.6%) was of course most common yeast in human sputums and the other species were seen in few.

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Molecular Cloning and Expression of Cellulase of Gene of Pseudomonas sp. in Escherichia coli (Pseudomonas sp.의 Cellulase 유전자의 대장균에의 클로닝 및 발현)

  • 정영철;김양우;노종수;성낙계;강신권
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.633-639
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    • 1990
  • The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.

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Purification and Immunological Properties of Vitellin, and its Quantitative Changes during Embyrogenesis in the silkworm, Bombyx mori (누에 난황소(Vitellin)의 분리와 면역학적 특성 및 발육에 따른 함량변화)

  • 손기국;문재유
    • Journal of Sericultural and Entomological Science
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    • v.30 no.2
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    • pp.96-104
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    • 1988
  • Vitellin, the major yolk protein of the silkworm, Bombyx mori was pruified, and its immunological properties and the quantitative changes during embryogenesis were studied. The ovary transplantation into male hosts was also carried out to find its effect on the yolk protein synthesis. The pupal vitellogenin and the egg vitellin of Bombyx mori were purified by DEAE-cellulose column chromatography. These two female specific proteins showed the same mobility in the polyacrylamide gel electrophoresis and the same reaction in the double immunodiffusion test. The immunological identity was also observed between the vitellins of Bombyx mori and Bombyx mandarina. The rudimentary ovaries transplanted into the male hosts of silkworms produced eggs without vitellin, indicating that the yolk precursors synthesized in other female organ beyond the ovary were necessary to produce vitellins. The major yolk protein, vitellin was disintegrated and utilized mostly during late stage of embryogenesis. It was different characteristics from the egg specific protein, which was utilized continuously from the early embryonic stage.

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Evaluation of Haemagglutinin Content by RP-HPLC to Generate Pandemic Influenza Vaccine

  • Kang, Hyunkyung;Roh, Hang Sik;Song, Hyemin;Lee, Kwangmoon;Chung, Seung-Tae;Ban, Sang-ja;Mo, In Pil;An, Beum-Soo;Ahn, Chi-Young
    • Toxicological Research
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    • v.32 no.4
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    • pp.269-274
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    • 2016
  • The potency of influenza vaccine is determined based on its hemagglutinin (HA) content. In general, single radial immunodiffusion (SRID) assay has been utilized as the standard method to measure HA content. However, preparation of reagents for SRID such as antigen and antibody takes approximately 2~3 months, which causes delays in the development of influenza vaccine. Therefore, quantification of HA content by other alternative methods is required. In this study, we measured HA contents of H1N1 antigen and H1N1 influenza vaccine by reverse phase-high performance liquid chromatography (RP-HPLC) methods. The presence of HA1 and HA2 was investigated by silver staining and Western blot assay. In addition, accuracy and repeatability of HA measurement by RP-HPLC were evaluated. Comparison of HA concentration by SRID and RP-HPLC revealed a precise correlation between the two methods. Our results suggest that RP-HPLC assay can replace SRID in the event of a pandemic flu outbreak for rapid vaccine development.

Characterization and Partial Nucleotide Sequence of Potato Virus X Isolated from Potato in Korea

  • Jung, Hyo-Won;Yun, Wan-Soo;Seo, Hyo-Won;Hahm, Young-Il;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.16 no.2
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    • pp.110-117
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    • 2000
  • Potato virus X (PVX-KO) showing mild mosaic and stunting symptoms on potato (Solanum tuberosum) in Kangwon area has been isolated and characterized. EM observation of the purified virus particles showed flexuous rod shape of about 520 nm in length. The coat protein (CP) of the virus had a molecular weight of 31 kDa in SDS-PAGE analysis, and the viral RNA was approximately 6.4 kb in size in denatured agarose gel electro-phoresis. In gel-immunodiffusion tests, it reacted strongly with an antiserum to common PVX from BIOREABAAG (USA). A rabbit antiserum was produced using purified virus and used for routine PVX detection by ELISA. Cultivated potatoes in Kangwon and other areas were frequently infected with PVX-KO. Both Datura stramonium and Nicotiana tabaccum cultivars developed necrotic local lesions 5 days after inoculation, and systemic mosaic symptoms with vein clearing 2 weeks after inoculation. All the features agree with the description of other PVX strains. To confirm and determine PVX strains, reverse transcription-polymerase chain reaction experiment was conducted using specific primers for viral CP. Amplified DNA fragments were cloned and sequenced. Results showed nucleotide sequence homologies of about 88 to 99% to other PVX strains. Based on CP amino acid sequence deduced from nucleotide sequences and host range studies PVX-KO is considered a member of the type X subgroup of PVX.

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The Production of Egg Yolk Immurnoglobulin (IgY) Raised against 3T3L-1 Cell Membrane Protein and the Control of Adipocytes Differentiation (3T3L-1세포의 막단백질에 대한 난황면역글로뷸린 (IgY)의 생산과 지방세포의 분화조절작용)

  • 김상윤;황성구;구의섭;고태송
    • Korean Journal of Poultry Science
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    • v.26 no.3
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    • pp.179-188
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    • 1999
  • The present was undertaken to establish a model for the control of adipocytes differentiation by using antibody from egg yolk. The emulsion of membrane protein of 3T3L-1 cell membrane protein with the complete Freund's adjuvant was firstly immunized in layer. Second and third boosting were undertaken with two weeks intervals by injection of the emulsion of the same antigen with the incomplete Freund's adjuvant. After 4 week of the first immunization, eggs were collected and antibody (IgY) was purified from egg yolk. The purity of IgY was 60-98% determined by single radial immunodiffusion (SRID) methods. Titer value of the antibody showed high reactiviy for the preadipocytes membrane protein measured by ELISA. When the IgY was added in the test media containing either 2.5% porcine serum or 10% FBS(control), the differentiation of 3T3L-1 cells and Glycerol-3-phosphate dehydrogenase(GPDH) activities was significantly decreased compared to the control cells(p〈0.05). When mice were subcutaneously injected with IgY raised against membrane protein of 3T3L-1 cells for 3 weeks, adipose tissue mass around ovary was tended to be decreased in female mice compared to those of control mice. It is suggested that a potential for manipulating of lipid accumulation through decrease in 3T3L-1 cell differentiation and fat accumulation in female mice by IgY treatment.

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SEROTYPES AND ANTIBIOTIC SUSCEPTIBILITIES OF STREPTOCOCCUS MUTANS ISOLATED FROM DENTAL PLAQUES OF CHILDREN (치아우식아동과 그의 모친(母親)의 치태(齒苔)에서 분리(分離)한 Streptococcus mutans의 혈청형(血淸型) 분포(分布) 및 약제내성(藥劑耐性)에 관(關)한 연구(硏究))

  • Baik, Byeong-Ju
    • Journal of the korean academy of Pediatric Dentistry
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    • v.11 no.1
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    • pp.75-89
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    • 1984
  • A total of 141 strains of Streptococcus mutans were isolated from dental plaques of 153 subjects. Eighty-two children with caries-experience, 18 caries-free children and their mothers were participated. All isolates were examined for their serotypes by immunodiffusion method with 7 reference sera and their antibiotic susceptibilities to 7 antibiotics by agar dilution method using 7 kinds of antibiotics, and were compared their results by caries experience (DMFT ratio) and by intrafamilial levels. 1. Isolation rate of Streptococcus mutans were greater in samples of carious teeth than those of sound teeth, and in mothers than in children. 2. Multiple serotypes of Streptococcus mutans were occasionally found in a single samples. 3. Of the total 141 isolates (83 isolates from children, and 58 from their mothers), type c isolates were most prevalent (63.8%). Type d,e and f were found, comprizing 14.9%, 10.6% and 6.4% respectively. Serotype g, a and untypable strain were also found but far lower frequencies (2.8-0.7%), and type b was detected. 4. These results suggest that there are no significant correlation among the distribution of serotypes, antibiotic susceptibilities, caries experience and intrafamilial relationships. 5. Most of isolates were susceptible to chloramphenicol (100%), penicillin (95.7%), ampicillin (94.3%), and gentamicin (92.2%), but about one-third isolates were resistant to cephaloridine (27%), streptomycin (33.3%) and kanamycin (47.6%), resulting that 91 strains (64.5%) among 141 isolates were resistant to one or more drugs used. 6. Of the 91 resistant strains, 20 different resistant patterns were observed, and the most frequently encountered patterns were KM, SM and CE.

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Identification of Pork Contamination in Meatballs of Indonesia Local Market Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis

  • Erwanto, Yuny;Abidin, Mohammad Zainal;Muslim, Eko Yasin Prasetyo;Sugiyono, Sugiyono;Rohman, Abdul
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.10
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    • pp.1487-1492
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    • 2014
  • This research applied and evaluated a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using cytochrome b gene to detect pork contamination in meatballs from local markets in Surabaya and Yogyakarta regions, Indonesia. To confirm the effectiveness and specificity of this fragment, thirty nine DNA samples from different meatball shops were isolated and amplified, and then the PCR amplicon was digested by BseDI restriction enzyme to detect the presence of pork in meatballs. BseDI restriction enzyme was able to cleave porcine cytochrome b gene into two fragments (131 bp and 228 bp). Testing the meatballs from the local market showed that nine of twenty meatball shops in Yogyakarta region were detected to have pork contamination, but there was no pork contamination in meatball shops in Surabaya region. In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity. Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked. The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

Production of Polyclonal Antibody against $\alpha$-Fetoprotein and Polyclonal Antibody-Based Competitive Enzyme-Linked Immunosorbent Assay for $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein (AFP)에 대한 폴리클로날 항체의 생산 및 $\alpha$-fetoprotein 측정용 효소면역분석법 (competitive ELISA)의 개발)

  • Michung Yoon
    • Biomedical Science Letters
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    • v.3 no.2
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    • pp.115-123
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    • 1997
  • $\alpha$-Fetoprotein (AFP) has been a useful marker in screening and/or monitoring patients with hepatocellular carcinoma, gonadal germ cell tumor, gastric carcinoma and neural tube defects. In the present study, it was attempted to produce anti-human AFP polyclonal antibodies and to develop a competitive enzyme-linked immunosorbent assay (ELISA) for the measurement of AFP in human plasma and amniotic fluid. AFP was isolated from amniotic fluid using an isolation procedure consisting of affinity chromatography and preparative polyacrylamide gel electrophoresis. The antibody directed against AFP was raised in rabbits. Double immunodiffusion and Western blotting methods showed that the antiserum was highly specific, reacting with only AFP-containing samples. Standard curve was obtained by using purified AFP and specific antiserum. The assay sensitivity was 5ng/ml and the working range was 5~l,000ng/ml. The within-assay and between-assay coefficient of variance (CV) was 4.5% and 8.5%, respectively. These results indicate that the assay is valuable for the measurement of AFP and found to be simple, reproducible, and accurate.

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THE CHANGES OF IMMUNOGLOBULIN ISOTYPES IN WHOLE SALIVA IN INFECTED PATIENTS OF ORAL AND MAXILLOFACIAL REGION (구강악안면 감염환자의 타액에서 Immunoglobulin Isotypes의 변화)

  • Byun, June-Ho;Chung, In-Kyo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.2
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    • pp.186-190
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    • 2000
  • Purpose : The purpose of this study is to observe the salivary immunoglobulin level in whole saliva of infected patients and also to investigate the changes of immunoglobulin level according to its management. Materials & Methods : Thirty infected patients who have been admitted to the dept. of oral and maxillofacial surgery of Pusan National University Hospital have been selected as subjects and we analysed the changes of immunoglobulin level of $1.5{\sim}3.0ml$ of unstimulated whole saliva collected throughout four times; the day before treatment, the first day after treatment, the third day after treatment and the day before discharge. We also compared them with immunoglobulins in whole saliva that was collected from 4 normal persons as control group. In radial immunodiffusion technique with BACKMAN(Array 360 system, McLean, USA), level of immunoglobulins was analyzed. Results : The isotypes of Ig that have been found in saliva of normal persons were IgG, IgA, IgM and IgE and their mean level was 8.23, 36.41, 4.38, and 2.38 respectively. In the infected patients before the treatment, the level of IgG, IgA was remarkably higher than that of normal persons, however we could not find the difference on the level of IgM, IgE. As the infection was healing, the level of IgG, IgA was decresing significantly.

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