• Restorative Dentistry and Endodontics
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• 제21권1호
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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.

• Journal of Acupuncture Research
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• 제31권1호
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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.

• 한국동물위생학회지
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• 제22권4호
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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.

• 한국축산식품학회지
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• 제40권1호
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• pp.132-144
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• 2020

The aim of this study was to optimize staining procedures for muscle fiber typing efficiently and rapidly in bovine and porcine skeletal muscles, such as longissimus thoracis, psoas major, semimembranosus, and semitendinosus muscles. The commercially available monoclonal anti-myosin heavy chain (MHC) antibodies and fluorescent dye-conjugated secondary antibodies were applied to immunofluorescence histology. Two different procedures, such as cocktail and serial staining, were adopted to immunofluorescence analysis. In bovine muscles, three pure types (I, IIA, and IIX) and one hybrid type, IIA+IIX, were identified by the cocktail procedure with a combination of BA-F8, SC-71, BF-35, and 6H1 anti-MHC antibodies. Porcine muscle fibers were typed into four pure types (I, IIA, IIX, and IIB) and two hybrid types (IIA+IIX and IIX+IIB) by a serial procedure with a combination of BA-F8, SC-71, BF-35, and BF-F3. Unlike for bovine muscle, the cocktail procedure was not recommended in porcine muscle fiber typing because of the abnormal reactivity of SC-71 antibody under cocktail procedure. Within the four antibodies, combinations of two or more anti-MHC antibodies allowed us to distinguish pure fiber types or all fiber types including hybrid types. Application of other secondary antibodies conjugated with different fluorescent dyes allowed us to get improved image resolution that clearly distinguished hybrid fibers. Muscle fiber characteristics differed depending on species and muscle types.

• Clinical and Experimental Reproductive Medicine
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• 제31권4호
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• pp.201-207
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• 2004

Objective: The Id family of helix-loop-helix proteins are thought to affect the balance between cell growth and differentiation by negatively regulating the function of basic-helix-loop-helix (bHLH) transcriptional factors. The aim of this study was to investigate the expression pattern of Ids (Id-1, -2, -3, and -4) in preimplantation mouse embryos at mRNA and protein levels. Methods: Oocytes and preimplantation embryos were collected from reproductive organs of female ICR mice following superovulation. RT-PCR was performed to investigate the mRNA expression patterns of Id genes and their protein were localized by immunofluorescence analysis. Results: Id-1 and Id-3 mRNAs were strongly expressed at the germinal vesicle (GV) oocyte and the blastocyst stages. Id-2 mRNA was expressed throughout preimplantation embryo development, but Id-4 was not expressed. Immunofluorescence showed that Id-1 and Id-2 were predominantly localized in cytoplasmic region, but the immunofluorescence signal of Id-3 was weak throughout preimplantation embryo development. Conclusion: These data show for the first time that Ids are expressed in preimplantation mouse embryos and suggest that Ids may play an important role in early preimplantation embryo development and uterine physiological changes.

• 한국식품과학회지
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• 제28권3호
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• pp.566-572
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• 1996

본 연구에서는 숙성 중 야기되는 식육의 인화정도를 파악할 수 있는 척도로서 이용하기 위하여, 근원섬유 Z선의 구성단백질의 하나인 zeugmatin에 대한 항체를 조제, 간접연역형광법으로 zeugmatin의 숙성중 변화와 근원섬유의 소편화와의 관계를 알아 보았다. Zeugmatin은 변성하기 쉬운 단백질로서 재래식 방법으로는 정제가 블가능하므로 이 단백질을 함유하는 획분을 정제과정 중에 채취하여 전기영동으로 전개, 이에 해당하는 band를 분리하여 polyclonal항체를 용이하게 받을 수 있었으며, 이 조제된 항체는 zeugmatin과 특이적으로 반응하였다. 조제된 항체를 이용하여 간접면역형광법으로 식육의 숙성중에 zeugmatin의 변화와 근원섬유의 소편화로 나타나는 식육의 연화와의 상관관계를 알아 본 결과, 이 두가지 변화는 특이적으로 일치하였다 즉, 닭의 흉근을 $4^{\circ}C$에서 숙성하면서 측정한 근원섬유의 소편화도는 도살 후 6시간에서 24시간 사이에 급속히 증가하여 이 시간대에 식육이 급속히 부드러워짐을 나타내었고, zeugmatin에 대한 항체가 나타내는 형광광도도 도살 후 6시간에서 24시간 사이의 동일한 시간대에 급적히 약화되어 zeugmatin이 이 시간대에 급속히 인화하였음을 나타내었다. 이상의 결과로 근원섬유 Z선의 구성단백질 중 하나인 zeugmatin은 숙성에 따라 야기되는 식육의 연화정도, 즉 식육의 숙성도를 나타내는 척도로서 이용될수 있으며, 그 이용방법으로는 간접면역형광법이 가장 간편한 방법인 것으로 판단되었다.

• 대한미생물학회지
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• 제21권3호
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• pp.393-397
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• 1986

Chlamydia trachomatis is one of the most common cause in non-gonococcal urethritis. There are several diagnostic methods for Chlamydia trachomatis; culture method using McCoy cell, enzyme immunoassay and direct immunofluorescence staining etc. We have studied a sensitivities of culture, chlamydiazyme and direct immunofluorescence staining(DIF). 85 male patients previously conformed to non-gonococcal urethritis have been selected in this study. Three samples were concurrently collected in the same patient. First sample was used to inoculation in McCoy cell, 2'nd sample was used to Chlamydiazyme test and 3'rd sample was used to direct immunofluorescence staining method. The results are following. 1. All culture, Chlamydiazyme and DIF positive cases are 15/85(17.7%). 2. Culture and Chlamydiazyme positive but DIF negative cases absent. 3. Culture and DIF positive, but Chlamydiazyme negative cases are 2/85(2.4%). 4. Chlamydiazyme and DIF positive, but culture negative cases are 9/85(10.6%). 5. Culture positive, but Chlamydiazyme and DIF negative cases are 6/85(7.1%). 6. Chlamydiazyme positive, but culture and DIF negative cases are 7/85(8.2%). 7. DIF positive, but culture and Chlamydiazyme negative cases are 3/85(3.5%). 8. All culture, Chlamydiazyme and DIF negative cases are 43/85(50.1%). In summarized, anyone positive cases of culture, Chlamydiazymc and DIF are 42/85(49.9%).

• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1621-1628
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• 2015

Chlamydia possesses a conserved 7.5 kb plasmid that is known to play an important role in chlamydial pathogenesis, since some chlamydial organisms lacking the plasmid are attenuated. The chlamydial transformation system developed recently required the use of plasmid-free organisms. Thus, the generation and identification of plasmid-free organisms represent a key step in understanding chlamydial pathogenic mechanisms. A tricolor immunofluorescence assay for simultaneously detecting the plasmid-encoded Pgp3 and whole organisms plus DNA staining was used to screen C. muridarum organisms selected with novobiocin. PCR was used to detect the plasmid genes. Next-generation sequencing was then used to sequence the genomes of plasmid-free C. muridarum candidates and the parental C. muridarum Nigg strain. We generated five independent clones of plasmid-free C. muridarum organisms by using a combination of novobiocin treatment and screening plaque-purified clones with anti-Pgp3 antibody. The clones were confirmed to lack plasmid genes by PCR analysis. No GlgA protein or glycogen accumulation was detected in cells infected with the plasmid-free clones. More importantly, whole-genome sequencing characterization of the plasmid-free C. muridarum organism and the parental C. muridarum Nigg strain revealed no additional mutations other than loss of the plasmid in the plasmid-free C. muridarum organism. Thus, the Pgp3-based immunofluorescence assay has allowed us to identify authentic plasmid-free organisms that are useful for further investigating chlamydial pathogenic mechanisms.

• Journal of Microbiology
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• 제38권1호
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• pp.24-30
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• 2000

US3 gene is a member of the human cytomegalovirus (HCMV) immediate early gene. Although the precise functions of the US3 gene in HCMV replication and pathogenesis are not known, it has been reported to play a role in inhibiting major histocompatibility class I antigen presentation. For further knowledge of US3 gene expression, rabbit polyclonal antiserum of the US3 gene product was used for indirect immunofluorescence assay. In permissive human foreskin fibroblast (HFF) cells, US3 gene expression was detectable as crescent or half-moon shape in the perinuclear region at immediate early times after virus infection. HFF cells infected with mutant HCMV lacking US3 open reading frames were negative for US3 immunofluorescence assay. Double immunofluorescence assay using monoclonal antibody to gamma adaptin (specific for the Golgi complex) and rabbit anti-US3 antiserum revealed that US3 gene product could be localized to the Golgi complex. At later time after HCMV infection, US3 gene products were detected as globular aggregates in the cytosol. These aggregates were positive for gamma adaptin and stained with preimmune serum, suggesting a nonspecific reaction to the Golgi complex. Northern blot analysis revealed that transcription of US3 was observed only during immediate early times after virus infection (until 6 h postinfection). Therefore US3 gene expression appears to be confined to immediate early time and its gene products are localized to the Golgi complex as crescent shaped forms in the perinuclear cytoplasm.

• Journal of the Optical Society of Korea
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• 제18권5호
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• pp.551-557
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• 2014

In this study, we demonstrated multimodal nonlinear optical (NLO) microscopy integrated simultaneously with two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), and coherent anti-Stokes Raman scattering (CARS) in order to obtain targeted cellular and label-free images in an immunofluorescence assay of the atherosclerotic aorta from apolipoprotein E-deficient mice. The multimodal NLO microscope used two laser systems: picosecond (ps) and femtosecond (fs) pulsed lasers. A pair of ps-pulsed lights served for CARS (817 nm and 1064 nm) and SHG (817 nm) images; light from the fs-pulsed laser with the center wavelength of 720 nm was incident into the sample to obtain autofluorescence and targeted molecular TPEF images for high efficiency of fluorescence intensity without cross-talk. For multicolor-targeted TPEF imaging, we stained smooth-muscle cells and macrophages with fluorescent dyes (Alexa Fluor 350 and Alexa Fluor 594) for an immunofluorescence assay. Each depth-sectioned image consisted of $512{\times}512$ pixels with a field of view of $250{\times}250{\mu}m^2$, a lateral resolution of $0.4{\mu}m$, and an axial resolution of $1.3{\mu}m$. We obtained composite multicolor images with conventional label-free NLO images and targeted TPEF images in atherosclerotic-plaque samples. Multicolor 3-D imaging of atherosclerotic-plaque structural and functional composition will be helpful for understanding the pathogenesis of cardiovascular disease.