• Restorative Dentistry and Endodontics
• /
• v.21 no.1
• /
• pp.121-135
• /
• 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.

• Korean Journal of Food Science and Technology
• /
• v.28 no.3
• /
• pp.566-572
• /
• 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.

• Restorative Dentistry and Endodontics
• /
• v.19 no.2
• /
• pp.485-498
• /
• 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.

• Microbiology and Biotechnology Letters
• /
• v.23 no.1
• /
• pp.110-117
• /
• 1995

This study was carried out to immunologically characterize Bacillus thuringiensis (B.t) antigens. Protein patterns of ultrasonicated- antigens of B. thuringiensis subspecies using SDS- PAGE revealed marked similarities among all the strains analyzed except for the difference between quantative variations of bands and some protein antigens. The comparison of the protein patterns showed that the protein antigen of 45 kilodalton (kd) was common in 11 strains and that the difference between B. thuringiensis subsp. canadensis and galleriae was noticed in quantative variations of bands despite of ambiguous serogrouping, suggesting a useful method for identification. All strains examined showed similar antigenic patterns in SDS-PAGE, while immunodominant bands differed in antigenic reactivity in western blot using polyclonal antibodies. Polyclonal antibody to B. thuringiensis subsp. thuringiensis and israelensis in indirect immunofluorescence assay reacted with flagella and cell surface antigens. The present study indicates that SDS-PAGE and western blot analysis may be used as tools for differentiation and identification of B. thuringiensis subspecies.

• Restorative Dentistry and Endodontics
• /
• v.24 no.1
• /
• pp.1-12
• /
• 1999

Bacterial infection of the pulp results in the development of a periapical lesion with the concomitant resorption of periapical bone. The cytokines are believed to play an important role in this matter. The purpose of this study was to find the relationship among the presence of black pigmented bacteria, the levels of cytokines(TNF-${\alpha}$, -${\beta}$, IL-$1{\beta}$, and TGF-${\beta}1$), and the amount of bone resorption in periapical and pulpal diseases. For the purpose, the patients were grouped into chronic apical pathosis, acute apical pathosis, acute pulpitis, and a healthy control group. Root canal samples were taken from periapical tissue exudates during routine endodontic treatment, and the venous blood was taken from each patients. The samples were processed to measure local and systemic levels of the cytokines using enzyme linked immunosorbent assay(ELISA). Bacterial content of Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella nigrescens were measured by indirect immunofluorescence method and the size of the periapical lesions were measured from the radiographs. The following results were obtained: 1. The levels of bone resorptive cytokines(TNF-${\alpha}$, TNF-${\beta}$, and IL-$1{\beta}$) in exudates from acute and chronic apical pathoses were significantly higher than those from acute pulpitis and the normal pulps(p<0.05). 2. IL-$1{\beta}$ were the highest among the bone resorptive cytokines in apical pathoses. However, no statistical difference between acute and chronic lesions were found(p>0.05). 3. The levels of TGF-${\beta}1$ in exudates from acute pulpitis and chronic apical pathoses were significantly higher than those from acute apical pathoses and the normal pulps(p<0.05). However, there were no significant correlations among the levels of bone resorptive cytokines. 4. The levels of TNF-${\beta}$ in serum were significantly higher than those from the exudates while serum TGF-${\beta}1$ concentrations were significantly lower(p<0.05). 5. Exudates from the canals in which the P. nigrescens were detected showed significantly higher levels of IL-$1{\beta}$ than those from the canals without the microorganism(p<0.05). 6. There were no significant correlations among the levels of the cytokines, the amount of bone destruction, and the presence of acute and chronic symptoms(p>0.05).

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.1
• /
• pp.21-26
• /
• 1997

This study was undertaken to examine the expression of EGF-R protein on ICM obtained from mouse IVF/IVC blastocyst by immunosurgery and indirect immunofluorescence(IIF). ICM cells used for this experiment were obtained from immunosurgery of mouse blatstocysts produced at 96 h after IVF, and recovered ICMs were assayed for cell viability and expression of EGF-R. The results obtained in this experiment were summarized as follows: when blastocysts were exposed to rabbit anti-mouse serum (antiserum) for 15-30 min. and then transferred them to guinea pig serum (complement) for 15-60 min., recovery rates of isolated ICMs were 8.0-84.2%. Especially, the best recovery (84.2%) of ICMs was obtained when exposure time to antiserum and complement was 30 min. and 60 min., respectively. In addition, when viability of isolated ICMs after immunosurgery was assessed by live/dead staining method, in all groups viability (93.8-100.0%) of isolated ICMs were not damaged if separated from TE cell. Also, we detected the expression of EGF-R on ICM cell by IIF. Therefore, these results suggest that EGF-R expression on the ICMs can stimulate the higher usablity of exogenous EGF to improve the preimplantation embryo development in vitro.

• Clinical and Experimental Reproductive Medicine
• /
• v.47 no.3
• /
• pp.186-193
• /
• 2020

Objective: The effectiveness of Staphylococcus protein A (SPA) in improving the penetration ability of sperm and reducing antisperm antibody (ASA) titers in immunologically infertile males was evaluated. Methods: Seminal fluid samples were obtained from 15 infertile men, and ASA titers were assessed with the latex agglutination test. Identification of immunoglobulin (Ig) classes and characterization of the antigens involved in the immune response were performed using indirect immunofluorescence. Local ASAs typically present as a mixture of IgG and IgA classes. The capillary tube penetration method was used to assess the capability of spermatozoa to penetrate the cervical mucus (CM). Results: ASAs associated with the neck region of sperm showed a significantly lower migration distance in the CM of infertile females than ASAs associated with the head or tail segments. ASA-positive seminal fluid exhibited significant increases in the mean migration distance (2.6 ± 1.4 cm vs. 1.54 ± 1.1 cm, respectively; p< 0.001) and sperm concentration (174 ± 121.0 × 10³/mL vs. 101 ± 93.7 × 10³/mL, respectively; p= 0.033) after treatment with SPA compared to pre-treated samples. A significant reduction (p< 0.01) in the recorded ASA titer was detected. Conclusion: These results indicate that SPA can be used as a sorting regimen for insemination programs. However, further studies are warranted to assess its influence on pregnancy rate.

• Korean Journal of Veterinary Research
• /
• v.34 no.4
• /
• pp.805-813
• /
• 1994

Immunohistochemical study on the intestinal tissues obtained from the 21 pigs of the 14 terms in Korea in which the clinical and epidemiological features had indicated the possible outbreaks of porcine epidemic diarrhea(PED) was performed using the indirect immunofluorescence test and/or the immunoperoxidase method in order to detect PED viral antigens in the infected cells of the intestines, and histopathological features were described as well. By immunohistochemical analysis, PED viral antigens were detected in the epithelial cells covering the small intestinal villi and recognized slightly in the cells lining the colonic surface epithelium as well. Occasional fluorescence was also seen in a few intestinal crypt epithelium. On light microscopy, the piglets with PED showed marked villous atrophy and fusion, and severe enterocyte degeneration and desquamation. On the other hand, the older pigs more than 4 week old age was mild villous atrophy and fusion, severe villous epithelial cell proliferation, and moderate mononuclear cell infiltration.

• Korean Journal of Veterinary Service
• /
• v.29 no.1
• /
• pp.1-8
• /
• 2006

Porcine circovirus (PCV) is a small, nonenveloped virus that contains a single-stranded circular DNA genome of about 1.76 kb and belongs to the family circoviridae. The PCV-2 has been incriminated as the cause of post-weaning multisystemic wasting syndrome (PMWS) , an emerging disease in pigs. In the present study, a PCR assay was applied to detect PCV-2 in tissue samples. The presence of PCV-2 antigen in the porcine tissues was confirmed by indirect immunofluorescence (IIF) with PCV-2 specific monoclonal antibodies. And then DNA extracted from PCV-2 positive tissues was used as a template. One oligonucleotide primer suitable for PCR was selected from a published PCV-2 sequence (Genbank). Amplified PCR product was detected the same fragment lengths of 416 bp as a control. Based on these results, it was suggested that the PCR is a simple and sensitive method for support diagnostic purposes.

• Journal of Veterinary Science
• /
• v.23 no.5
• /
• pp.32.1-32.11
• /
• 2022

Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases. The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions: The spbELISA has practical applications in assessing the vaccination status of large pig herds.