• Title/Summary/Keyword: 중합효소 연쇄반응

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Quantitative detection of peri-implantitis bacteria using real-time PCR (Real-time PCR을 이용한 임플란트주위염 원인균의 정량적 분석)

  • Kim, Min-Jung;Han, Gyeong-Soon
    • Journal of Korean society of Dental Hygiene
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    • v.21 no.5
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    • pp.555-565
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    • 2021
  • Objectives: This study was conducted to analyze peri-implantitis bacteria and identify their associations with health status and health activities. Methods: Gingival sulcus fluid at the implant's periodontal pockets sampled from the participants were analyzed by multiplex real time PCR. Results: Participants had strains in the order of 100% F. nucleatum, 98.0% E. corrodens, and 96.0% P. micra, and the correlation between C. rectus and E. nodatum was high (p<0.01). Diabetic group (P. gingivalis, P. nigrescens) hypertension (P. nigrescens), group with four or more periodontal pockets (P. gingivalis, T. dentica, P. intermedia, E. nodatum, and C. rectum), smoking (P. micra, E. corrodens), drinking (T. dentola), and scaling groups (C. rectus) were found to have more strains (p<0.05). Conclusions: Representative pathogenic microorganisms detected in periodontal pockets of implants were similar to dental periodontal pockets; however there were differences in the amount and distribution of microorganisms, and they were affected by health status and health behavior.

Detection of Viral Antigens in Stool Using EIA in Hospitalized Children and Clinical Implication (간접 효소 면역측정법을 이용한 입원 환아의 대변에서 바이러스 항원의 검출과 임상적 의의)

  • Min, Jung Hye;Seo, Jeong Wan;Park, Hye Kyung
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.7 no.2
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    • pp.143-152
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    • 2004
  • Purpose: The purpose of this study is to detect viral coproantigens in children who were hospitalized with acute diarrhea and to compare its association with clinical symptoms. Methods: Seventy-four stool samples were collected from children admitted to Ewha Mokdong Hospital from March 1996 to December 1999. The samples were frozen and analyzed for rotavirus, adenovirus, enterovirus, astrovirus, and calicivirus by enzyme immunoassay (EIA) with monoclonal antibody. 53 stool samples were collected from patients with diarrhea (diarrheal group) and 21 stool samples from patients hospitalized for reasons other than diarrhea (control group). Clinical features and laboratory findings were reviewed in both groups. Results: Among 74 stool samples, virus antigens were detected in 60 samples. Of the 60 virus-positive stool samples, 47 enterovirus, 26 rotavirus, 16 adenovirus, 11 astrovirus, and 11 calicivirus antigens were detected by EIA. Of the 60 virus-positive stool samples, 28 samples have one viral antigen, 30 samples have 2 or more viral antigens, and 2 samples showed a simultaneous infection of Salmonella group B and enterovirus. There was no relationship between the detected virus and clinical features. Conclusion: In this study, viral coproantigen and clinical symptoms were not associated. In the future, further larger scale studies are necessary.

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Use of Nested Polymerase Chain Reaction for Identification of Rickettsia tsutsugamushi Serotype Cultured in Human Embryonic Lung Cells (Nested PCR을 이용한 사람 유래 태아 폐세포에서 배양된 Rickettsia tsutsugamushi의 혈청형 동정)

  • An, Chang-Nam;Woo, Gyu-Jin;Kim, Tae-Yeon;Shin, Kwang-Soon;Kim, Chul-Joong;Baek, Luck-Ju
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.235-244
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    • 1996
  • We selected the adequate cell line to be used for propagation and plaquing of R. tsutsugamushi in laboratory and identified R. tsutsugamushi serotype cultured in LuMA cells by nested PCR. As in this study, we concluded that. 1. LuMA cell was suitable for the study of the biology of rickettsiae-host cell interaction. 2. The plaque-forming unit (PFU) per ml of R. tsutsugamushi Karp strain propagated in embryonated egg yolk sacs was $10^{8.8}$ and the PFU/ml of Gilliam strain was $10^{7.1}$. 3. The rate and extent of cytopathic changes depended on the PFU titer of R. tsutsugamushi. 4. PCR with nested primer pairs was useful for identification of R. tsutsugamushi serotype cultured in human embryonic lung cells.

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Characterization of hrp2 + Gene Related to SNF2 Family in Schizosaccharomyces pombe (Schizosaccharomyces pombe에서 SNF2에 속하는 hrp2+ 유전자의 특성 연구)

  • Park, In-Soon
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.137-141
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    • 2002
  • The SNF2/SW12 family comprises proteins from a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. This study was shown the characterization of hrp2+ gene which was isolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of hrp2+ gene showed striking evolutionary conservation among the SNF2 family of proteins. The transcript of hrp2+ gene was found to be a 4.7 kb as identified by Northern hybridization. In addition, to determine the transcription initiation site of hrp2+ gene, primer extension analysis was performed. This result showed the band of 64 bp. The transcriptional start point was mapped to a position of 47 base pair from the first ATG codon of translational initiation codon. In order to investigate the inducibility of hrp2+ gene, transcript levels were examined after treating the cells to various DNA damaging agents. The transcripts of hrp2+ were induced by UV-irradiation. But the transcripts were not induced by treatment of 0.25% Methylmethane sulfonate (MMS). These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of this gene.

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Study on Microbiochip for Buccal Cell Lysis and DNA Purification (상피세포 시료 전처리용 마이크로바이오칩에 관한 연구)

  • Ha, Seung-Mo;Cho, Woong;Ahn, Yoo-Min;Hwang, Seung-Yong
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.34 no.12
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    • pp.1785-1791
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    • 2010
  • This paper describes a separable microfluidic device fabricated with PDMS (polydimethylsiloxane) and glass. The device is used for sample preparation involving cell lysis and the DNA purification process. The cell lysis was performed for 2 min at $80^{\circ}C$ in a serpentine-type microreactor ($20 {\mu}l$) using a Au microheater that was integrated with a thermal microsensor on a glass substrate. The DNA that was mixed with other residual products during the cell lysis process was then filtered through a new filtration system composed of microbeads (diameter: $50 {\mu}m$) and PDMS pillars. Since the entire process (sample loading, cell lysis reaction, DNA purification, and sample extraction) was performed within 5 min in a microchip, we could reduce the sample preparation time in comparison with that for the conventional methods used in biochemistry laboratories. Finally, we verified the performance of the sample preparation chip by conducting PCR (polymerase chain reaction) analysis of the chip product.

Relationship Between Mitochondrial DNA Copy Number, Metabolic Abnormalities and Hepatic Steatosis (지방간 및 대사 인자들과 말초혈액 백혈구의 사립체 DNA copy 수와의 연관성)

  • Kwon, Kil-Young;Jun, Dae-Won
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.6
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    • pp.2093-2098
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    • 2010
  • Insulin resistance plays a central role in fatty liver, a part of the metabolic syndrome. This study examined the relationship between fatty liver, metabolic abnormalities and mitochondrial DNA [mtDNA] copy number in peripheral blood that is correlated with diabetes or metabolic markers. Fatty liver was assessed by questionnaire on alcohol consumption and abdominal ultrasonography. MtDNA copy number in peripheral leukocytes was measured by a real-time quantitative polymerase chain reaction [PCR]. Among 445 subjects, 148 subjects had hepatic steatosis and 297 were controls. mtDNA copy number was significantly lower in fatty liver group in comparison with that of normal finding group. This result is similar in both groups, alcoholic or non-alcoholic fatty liver group. MtDNA copy number was inversely correlated with alanine aminotransferase [ALT], aspartate aminotransferase [AST], gamma-glutamyltransferase [$\gamma$-GTP], body mass index [BMI], waist circumference, diastolic blood pressure, and free fatty acid. MtDNA copy number in peripheral leukocytes was associated with fatty liver and insulin resistance related factors.

Isolation and Characterization of hrp2+ Gene Related to SNF2 Family In Yeast (Schizosaccharomyces pombe에서 SNF2에 속하는 hrp2+ 유전자의 특성 연구)

  • Choi In Soon
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.192-196
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    • 2005
  • The SNF2/SW12 family comprises proteins from a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. This study was shown the characterization of hrp2+ gene which was isolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of hrp2+ gene showed striking evolutionary conservation among the SNF2 family of proteins. The transcript of hrp2+ gene was found to be a 4.7 kb as identified by Northern hybridization. To investigate the inducibility of hrp2+ gene, transcript levels were examined after treating the cells to various DNA damaging agents. The transcripts of hrp2+ were induced by UV-irradiation. But the transcripts were not induced by treatment of $ 0.25\%$ Methylmethane sulfonate (MMS). These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of this gene. Hrp2 protein was purified near homogeneity by combination of affinity chromatography. We tested the purified Hrp2 protein for the helicase activity in an oligonucleotide release assay. However we were unable to detect any helicase activity associated with the Hrp2 protein, indicating that the helicase motifs in Hrp2 are merely indicators of a broader DNA-dependent ATPase activity.

Study on PPARγ-dependent adipogenesis regulation by testosterone using transient transfection assays (일시적인 형질전환 분석을 이용한 testosterone에 의한 PPARγ-의존적 지방세포생성 조절에 관한 연구)

  • Jeong, Sunhyo
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.18 no.2
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    • pp.482-487
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    • 2014
  • This study is to investigate the effects of testosterone on adipogenesis and its molecular mechanism using RT-PCR analysis and transient transfection assays. Castrated(CAST) mice treated with testosterone had lower white adipose tissue weights and expression of adipocyte-specific genes($PPAR{\gamma}$ and aP2) than CAST control mice. Consistent with the in vivo data, testosterone treatment inhibited triglyceride accumulation and expression of adipocyte-specific genes($PPAR{\gamma}$ and aP2) in differentiated 3T3-L1 cells compared with control group. Testosterone-activated androgen receptor(AR) repressed the luciferase reporter gene activity induced by $PPAR{\gamma}$ transfection. Thus, these results suggest that testosterone downregulates the actions of $PPAR{\gamma}$ on adipogenesis through AR.

Investigation of effect of zirconia on osseointegration by surface treatments (지르코니아 표면처리가 골유착에 미치는 영향)

  • Jeong, Jin-Woo;Song, Young-Gyun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.37 no.1
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    • pp.23-30
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    • 2021
  • Purpose: The aim of this study was to investigate effect of zirconia on osseointegration and Surface appearance by surface treatments using various acid solution. Materials and Methods: The prepared zirconia disks were treated with hydrofluoric acid solution and photo-assisted etching under various condition. The surface was analyzed by SEM and the surface roughness was analyzed by using surface profiler. The osteogenic effect of MC3T3-E1 cells was assessed via fluorescent staining observation and reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Various roughness were obtained according to the surface treatment method. The surface roughness increased in the group treated with hydrofluoric acid solution, but that had week network structure. In the method using photo-assisted etching, the surface roughness increased in micro units. Cell reaction showed better results in the photo-assisted etching group than in the hydrofluoric acid-treated group (P < 0.05). And it showed even osteoblastic cell distribution in photo-assisted etching group. Conclusion: As a result, the photo-assisted etching method is more effective than the simple acid solution treatment for zirconia treatment for osseointegration.

Anti-inflammatory Effect of Ishige foliacea in RAW 264.7 Cells (넓패추출물에 의한 RAW 264.7 세포에서의 항염효과)

  • Joonghyun Shim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.1
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    • pp.29-36
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    • 2024
  • This study was carried out to identify the anti-inflammatory effects of Ishige foliacea (I. foliacea) extract on skin using RAW 264.7 cells. The anti-inflammatory effects of I. foliacea extract on RAW 264.7 cells were assessed by cell viability assay, mRNA expressions, and nitric oxide (NO)/prostaglandin E2 (PGE2) productions. The anti-inflammatory effects of I. foliacea extract were elucidated by analysis of IL-1α/IL-1β/IL-6/TNFα gene expressions and PGE2/NO production. Quantitative real-time polymerase chain reaction showed that I. foliacea extract decreased the gene expression levels of iNOS/COX2/IL-1α/IL-1β and IL-6. Furthermore, PGE2/NO production also revealed that I. foliacea extract exhibited anti-inflammatory properties. These results suggest that I. foliacea extract is an anti-inflammatory compound. It could be a potent cosmeceutical material for anti-inflammatory effects. Further studies on the anti-inflammatory mechanisms of broadleaf extracts are expected to help identify pharmacological mechanisms related to inflammation in addition to cosmeceuticals.