• Title/Summary/Keyword: Indirect Immunofluorescence

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INDIRECT IMMUNOFLUORESCENCE FOR THE IDENTIFICATION OF ACTINOMYCES SPECIES IN PATIENTS WITH PERIAPICAL LESION (치근단 병변에서 Actinomyces 종의 검정을 위한 간접 면역형광법적 연구)

  • Chang, Won-Jung;Yoon, Soo-Han;Kwon, O-Yang
    • Restorative Dentistry and Endodontics
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    • v.21 no.1
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    • pp.121-135
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    • 1996
  • Actinomyces are Gram-positive, non-acid-fast, anaerobic or microaerophilic filamentous bacteria. These organisms are frequently detected from infected root canals and periapical lesion. The purpose of this study was to use indirect immunofluorescence to determine the prescence of select Actinomyces species in a survey of teeth associated with periapical lesion, to clarify the relationship between clinical symptoms of periapical lesions and the Actinomyces species and to study on the cross reaction among Actinomyces. Actinomyces israelii serotype I (ATCC 12102), Actinomyces israelii serotype II (ATCC 29322), Actinomyces viscosus serotype II (ATCC 19246), Actinomyces naslundii serotype I (ATCC 12104) were cultured in anaerobic condition. Rabbit antisera were prepared by intravenous injection of formalized whole cells. Indirect immunofluorescence method was used to achieve the purpose. The following results were obtained. 1. There was a relationship between Actinomyces and periapical disease. 2. A. israelii serotype I, II were frequently identified with Indirect Immunofluorescence and most often assosiated with periapical disease. In culture finding, there was no significant difference between each group. 3. Indirect Immunofluoresence is both more sensitive and more rapid than culture for identification of Actinomyces species in patients with periapical lesion. 4. A. israelii serotype I, II was highly isolated in infected root canals with local swelling, A. naslundii serotype I was highly isolated in those with foul odor, and A. israelii serotype I was found in higher frequncy in those with exudate than other bacteria. 5. In the Indirect Immunofluorescence (1 : 320), A positive cross reaction was obtained between A. israelii serotype I and A. israelii serotype II, also, A. viscosus serotype II and A. naslundii serotype I. There was no cross reaction between A. israelii serotype I, II and A. viscosus serotype II, A. naslundii serotype I.

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Studies on the Characteristics of Anti-Zone Antibody I. Identification of Anti-Zona Antibody by Indirect ELISA and Indirect Immunofluorescence Test (항투명대 항체의 특성에 관한 연구 I. 간접효소면역분석법과 간접면역형광분석법에 의한 항투명대 항체의 확인)

  • 김은영;유영춘;박세필;정형민;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.107-114
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    • 1990
  • These experiments were undertaken as a basic study to develop immunocontraceptive vaccine and to understand the role of zona pellucidae in early fertilization process by identifying the monoclonal and polyclonal antibody to porcine zona pellucidae and polyclonal antibody to mouse zona pellucidae by indirect ELISA and indirect immunofluorescence test. The results obtained in these experiments were summarized as follows : 1. The titer of the antibodies to zona was determined by indirect ELISA using solubilized porcine zona coated plates. Both monoclonal and rabbit polyclonal antibodies showed very high titers ; O.D at 1 : 12,800 dilution of antibodies was still significantly higher than that of non-immunized control serum. Rabbit anti-mouse zona pellucidae sera also reacted with porcine zona pellucidae. 2. By indirect immunofluorescence test strong fluorescences were observed on the egg treated with homologous and heterologous rabbit polyclonal antibodies and FITC lablled 2nd antibodies and found to crossreact strongly with the eggs from the pig and mouse. While weaken fluorescences were observed on the eggs treated with monoclonal antibodies.

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Effects of Indirect Moxibustion on Skeletal Muscles in Mouse Model of Skeletal Muscle Adiposity (간접구 시술이 골격근 Adiposity 유발 쥐의 근육조직에 미치는 영향)

  • Lee, Ki Su;Hong, Kwon Eui
    • Journal of Acupuncture Research
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    • v.31 no.1
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    • pp.7-21
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    • 2014
  • Objectives : To observe the regenerative effects of indirect moxibustion, a traditional Korean medical treatment on skeletal muscles using mouse model of skeletal muscle adiposity. Methods : Twenty seven ICR male mice were randomly assigned into Intact control(n=3), glycerol treatment together without moxibustion(n=12), and glycerol treatment together with moxibustion (n=12) groups. Mice of glycerol treatment groups were injected with 50 ${\mu}l$ DW(distilled water) containing 50 % of glycerol into the two tibialis anterior. After injection, moxibustion was applied at 'Shenshu'($BL_{23}$) and 'Zusanli'($ST_{36}$) acupoints three times per each session, every days for twelve days(total 12 treatments). Phospho-Erk1/2, Myostatin protein levels were analyzed by western blotting and immunofluo-rescence staining techniques for tissues of the tibialis anterior muscle. Smad, phospho-Smad were analyzed by immunofluorescence staining. Results : 1. Histological analysis of sections from injected TA muscles showed that glycerol induced rapidly muscle necrosis, with a maximum at day 3. 6 days and 9 days after injection, muscle was regenerating. 2. According to western blotting and immunofluorescence staining, phospho-Erk1/2 protein signals in glycerol treatment with moxibustion group were stronger compared to Intact and glycerol treatment without moxibustion group. 3. According to western blotting and immunofluorescence staining, myostatin protein signals in glycerol treatment without moxibustion group were stronger compared to Intact and glycerol treatment with moxibustion group. 4. According to immunofluorescence staining, Smad protein signals in glycerol treatment without moxibustion group were stronger compared to Intact and glycerol treatment with moxibustion group. 5. According to immunofluorescence staining, phospho-Smad protein signals in glycerol treatment without moxibustion group were stronger compared to Intact and glycerol treatment with moxibustion group. Conclusions : These results confirm that indirect moxibustion of 'Shenshu'($BL_{23}$) and 'Zusanli'($ST_{36}$) influences muscle regeneration in mouse models of skeletal muscle adiposity. Further discussion, and the establishment of moxibustion mechanism will prompt clinical application of moxibustion.

Serological studies of avian rotaviruses in chicken sera using indirect immunofluorescence assay (간접형광항체법을 이용한 닭 로타바이러스의 혈청역가 조사)

  • 김혜경;곽학구;박경재;안재문;강신영
    • Korean Journal of Veterinary Service
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    • v.22 no.4
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    • pp.385-392
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    • 1999
  • A total of 420 chicken sera from various regions were tested for the presence of antibodies to avian rotavirus using indirect immunofluorescence assay (IFA). In broiler farms, rotavirus antibodies were detected from 20 farms among 30 farms tested and the positive rates were above 50% in 9 farms. In parent stock farms, rotavirus antibodies were detected from 5 farms among 14 farms tested. From sera collected in 7 layer farms rotavirus antibodies were not detected.

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Measurement of Meat Tenderization during Post-mortem Aging by the Indirect Immunofluorescence Method (간접면역형광법(間接免疫螢光法)을 이용(利用)한 숙성중(熟成中) 식육(食肉)의 연화정도(軟化定度) 측정(測定))

  • Ahn, Dong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.28 no.3
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    • pp.566-572
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    • 1996
  • The relationship between the myofibrillar fragmentation and zeugmatin during post-mortem aging was in vestigated by indirect immunofluorescence method using antizeugmatin Antiserum as a measure of meat tenderization. The antizeugmatin antiserum was prepared using bands separated by SDS-PAGE and reacted specifically with zeugmatin, showing no cross-reactivity with the other myofibrillar proteins. By the indirect immunofluorescence method, this antiserum stained the fresh myofibrillar However, the fluorescence intensity decreased with post-mortem time and almost disappeared within 24 hr of storage, in parallel with the myofibrillar fragmentation. It was therefore concluded that zeugmatin can be conveniently used as a measure of meat tenderization during post-mortem aging by immunoflurescence method.

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IMMUNE REACTION OF SPECIFIC SERUM ANTIBODIES TO PORPHYROMONAS ENDODONTALIS ANTIGEN IN PATIENTS WITH PERIAPICAL LESION (치근단 병소가 있는 환자에서 Porphyromonas endodontalis 항원에 대한 혈청 특이 항체의 면역 반응 연구)

  • Kim, Jae-Hee;Yoon, Soo-Han
    • Restorative Dentistry and Endodontics
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    • v.19 no.2
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    • pp.485-498
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    • 1994
  • Porphyromonas endodontalis is a black-pigmented anaerobic Gram-negative rod which is associated with endodontal infections and this microorganism possesses a potential for pathogenicity. The purpose of this study was to compare the membrane components of Porphyromonas endodontalis and Porphyromonas gingivalis and to study the immune reaction patterns of Porphyromonas endodontalis with patients with periapical lesion. Porphyromonas endodontalis (ATCC 35406), Porphyromonas gingivals serotypea (381), serotype b(W50), serotype c(A7A1-28) were cultured in anaerobic condition. Rabbit antisera were prepared by intravenous injection of formalized whole cells and human sera were obtained from patients and dental students. Indirect immunofluorescence method was used to study on the cross reaction between Porphyromonas endodontalis and Porphyromonas gingivalis serotype a, b, c antigen. Total membrane protein profiles of Porphyromonas endodontalis antigen were studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and the reactivity of antigenic components of Porphyromonas endodontalis against sera of patients and rabbit anti-Porphyromonas endodontalis antisera were assessed by Immunoblotting method. The following results were obtained : 1. Antigens of Porphyromonas endodontalis has multiple antigenic components, and both patients with periapical lesion and normal healthy individual showed immune response to this. 2. Patients group and healthy individual group showed a diversity of immune reaction pattern but they showed immune response against 43kd protein. 3. Patients with periapical lesion showed more diverse immune response than healthy individual and in some patients, much more bands appeared to lower molecular weight protein. 4. According to indirect immunofluorescence and Immunoblotting study, Porphyromonas endodontalis did not share common antigen with Porphyromonas gingivalis serotype a, b, c.

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Serological Study on the Cross-Reactivity of Bacteroides gingivalis, Bacteroides intermedius and Bacteroides asaccharolyticus by Indirect Immunofluorescence and Enzyme- Linked Immunosorbent Assay (형광 현미경법 및 효소결합 면역흡착법을 이용한 Bacteroides gingivalis, Bacteroides intermedius 및 Bacteriudes asaccharolyticus의 혈청학적 연구)

  • Chung, C.P.;Lee, J.Y.;Lee, Y.H.;Chung, H.W.;Chung, H.J.
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.117-123
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    • 1987
  • Previous studies have been performed for the sero-identification of selected species of Bacteroides by immunofluorescence antibody techniques and enzyme-linked immunosorbent assay using species-specific rabbit antisera to B. gingivalis, B. intermedius, and B. melaninogenicus. However, these studies have not commented on the serological cross-reactivity between these 3 species of black- pigmented Bacteroides. For the cross-reactivity study, antisera to B. gingivalis ATCC 33277, B. intermedius ATC C25261 and B. asaccharolyticus ATCC 25260 were raised from rabbits. Preliminary study for observing the cross-reactivity between these species was performed by indirect immunoflourescence technique. Immunoabsorption of the antisera was done with bacterial cells from the other species and the species-specificity of the antisera was conformed by the absence of reactivity with bacterial strains from the other species by indirect immunofluorescence technique and enzyme-linked immunosorbent assay. Three representative unabsorbed antisera cross-reacted strongly with cells from the other species. Especially, anti-B. asaccharolyticus ATCC 25260 antiserum showed a strong cross-reactivity with B. gingivalis ATCC33277. After immunoabsorption of 3 representative antisera with the other species, the cross-reactivity was found only between B. gingivalis ATCC 33277 and B. asaccharolyticus ATCC 25260. Further study would be necessary to clarify the cross-reactivity between important oral black-pigmented Bacteroides from subgingival plaque or bacterial colonies for rapid identification.

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Localization of a KEM1::lacZ Fusion Protein in Yeast Cells (효모세포에서 KEM1::lacZ 융합 단백질의 위치결정)

  • Kim, Jin-Mi;Fink, Gerald R.
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.12-19
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    • 1994
  • KEM1 is known to control the spindle pole body or microtubule function, probably in response to the cellular nutritional conditions in Saccharomyces cerevisiae. Transposon insertions were performed in the cloned KEM1 gene using mini-Tn10-LUK element carrying E. coli ${\beta}$-galactosidase structural gene. A collection of ranfom Tn10-LUK insertions defined an approximately 3.5 kb region required for the KEM1 function. From this collection functional KEM1::lacZ protein fusions were identified. Indirect immunofluorescence using anti-${\beta}$-galacatosidase antibodies localized the KEM1::lacZ fusion protein to the periphery of the nucleus.

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Evaluation of Anti-AFP Monoclonal Antibodies as Immunodiagnostic Reagents for Hepatocellular Carcinoma (항-AFP 단일클론 항체를 이용한 간암진단 효과의 검토)

  • 현광자;강희갑;강신성
    • The Korean Journal of Zoology
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    • v.38 no.3
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    • pp.382-387
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    • 1995
  • To check the possible application of our anti-AFP monocional antibodies (MAbs) as immunodiagnostic reagents for hepatocellular carcinoma, ELISA and immunohistochemical assay were performed on the sera and liver biopsy specimens from the patients of hepatocellular carcinoma and other non-malignant hepatic disease. By non-competitive ELISA using anti-AFP MAbs, the highest incidence of AFP value was found only in the sera of hepatocellular carcinoma patients, i.e., more than 54% of patients had serum AFP levels of more than 500 ng/mi. By immunoperoxidase and indirect immunofluorescence techniques, anti-AFP MAbs were found to react with cytoplasm of hepatoceliular carcinoma cells. However immunohistochemical reactIvity to AFP in hepatocellular carcinoma cells was lower than that in non-neoplastic liver cells adjacent to the hepatocellular carcinoma. From these results with the similar findings from other studies, we suggest that AFP antigen is appropriate in the diagnosis assay (ELISA) but is not by immunohistochemical detection.

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