• Title/Summary/Keyword: 실시간 중합 효소 연쇄 반응

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INHIBITION OF GLUCAN SYNTHESIS RELATED GENE EXPRESSION OF STREPTOCOCCUS MUTANS BY XYLITOL TREATMENT (자일리톨 섭취에 따른 Streptococcus mutans의 글루칸 생성관련 유전자 발현 억제효과)

  • Kim, Ji-Hye;Lee, Young-Eun;Ahn, Sang-Hun;Choi, Youn-Hee;Nam, Soon-Heyun;Song, Keun-Bae
    • Journal of the korean academy of Pediatric Dentistry
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    • v.36 no.4
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    • pp.531-538
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    • 2009
  • Xylitol has the ability to reduce the adherence of Streptococcus mutans(S. mutans), which can make it easier to remove plaque, decrease acid production and inhibit dental caries. There are few reports on the effects of xylitol on the expression of the virulence related genes in S. mutans. This study examined the inhibitory effect of chewing gum containing xylitol on glucan synthesis related gene expression of S. mutans. Participants were voluntarily recruited for a women's oral health prevention program, classified into two groups(a control and a xylitol group), and then followed for 2 years. Twenty salivary samples were randomly selected from each group. Colony count and real-time reverse transcription polymerase chain reaction were used to analyze the characteristics of S. mutans. The following results were obtained: The S. mutans counts decreased steadily in the xylitol group over the study period(p<0.05). The expression of the virulence related genes (gtfB, gtfC and gtfD) was significantly lower in the xylitol group than in the control groups (p<0.05). In conclusion, these results suggest that chewing xylitol gum for a long period of time may reduce the expression of the genes associated with S. mutans virulence, which can result in a decrease growth of S. mutans colonies as a result.

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Anti-inflammatory, Anti-aging, and Sebum Inhibitory Effects of Osmanthus fragrans Flower Extract (목서 꽃 추출물의 항염, 항노화 및 피지 억제 효능)

  • Hyung-Min Kim;Yeon Su Jeong;Sehyun Kim;Jeong Hun Cho;Yong Deog Hong;Won-Seok Park
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.2
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    • pp.171-178
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    • 2024
  • In this study, we aimed to determine the various effects of Osmanthus fragrans (O. fragrans) flower extract on the skin in order to utilize it as a cosmetic material. For this purpose, Osmanthus fragrans flower extract (OFFE) of Jeju Island was prepared and used in the experiment. The experiments were evaluated by the quantitative real-time polymerase chain reaction (qRT-PCR) and lipid droplet staining assay. First, the OFFE decreased the gene expressions of three representative pro-inflammatory cytokines (IL-8, IL-6, and IL-1α) and an inflammation-related enzyme, PTGS2 induced by poly I:C in epidermal keratinocytes. In addition, the OFFE increased the gene expression levels of collagen (COL1A1) and elastin (ELN) in dermal fibroblasts. Further, the OFFE showed the inhibitory effect in sebum production by linoleic acid in sebocytes. Therefore, from this study, it is expected that OFFE can be used as a natural cosmetic material for anti-inflammatory, anti-aging, and sebum inhibitory efficacy.

Characterization of Odontoblasts in Supernumerary Tooth-derived Dental Pulp Stem Cells between Passages by Real-Time PCR (과잉치 치수유래 줄기세포의 Real-time PCR에 의한 계대간 상아질모세포 발현 특성)

  • Ji, Sangeun;Song, Sol;Lee, Joonhaeng;Kim, Jongbin;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.48 no.3
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    • pp.291-301
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    • 2021
  • The aim of this study is to compare the properties of odontoblast gene of early passage cells and late passage cells derived from impacted maxillary supernumerary teeth. Impacted supernumerary teeth with maxilla were extracted from 12 patients (8 males, 4 females) between 6 - 9 years old without medical history. Real-time polymerase chain reaction (PCR) was conducted to compare characterization of odontoblast cell in the 3rd and 10th passage, and between with bone inducing additive group and without additive group. Genes for odontoblasts characteristics are osteonectin (ONT), alkaline phosphatase (ALP), osteocalcin (OCN), dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP). The level of gene expression was in a decreasing order of ONT, ALP, OCN, DMP-1 and DSPP in the 3rd passage, and in decreasing order of ONT, DMP-1, OCN, ALP, and DSPP in the 10th passage in the undifferentiation and differentiation group. The order of ONT, DMP-1, and OCN did not changed. ALP and DMP-1 were switched in order. ALP and DMP-1 may be used as important markers for differentiating between the 3rd passage and 10th passage cells. Considering that supernumerary tooth was extracted young age and the time required to cultured 10th passage was short, supernumerary tooth can be considered a useful donor site of dental pulp stem cells.

Development of Techniques for Evaluating the Virus Removal Rate using Adenovirus (아데노바이러스를 이용한 바이러스 제거율 평가를 위한 기법 개발)

  • Cho, Yoonjung;Lim, Jaewon;Baek, Dawoon;Lee, Sang-Hoon;Lee, In-Soo;Lee, Hyeyoung;Park, Donghee;Jung, Dongju;Kim, Tae Ue
    • Journal of Korean Society of Water and Wastewater
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    • v.29 no.6
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    • pp.633-641
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    • 2015
  • Waterborne infectious disease is induced by several pathogenic microbes such as bacteria, viruses and protozoans, and the cases caused by viral infection is currently increasing. Water treatment process could reduce the number of virus in the water, but there were many difficulties to completely remove the virus particles from water. Therefore, the membrane separation technology which was reported to effectively remove pollutants from raw water has attracted increasing attention and demand. Since its efficiency has been introduced, demands for evaluation method toward the membrane filtration process are increasing. However, progression of the method development is slow due to the difficulties in cultivation of several waterborne viruses from animal models or cell culture system. To overcome the difficulties, we used adenovirus, one of the commonly isolated pathogenic waterborne viruses which can grow in cell culture system in vitro. The adenovirus used in this study was identified as human adenovirus C strain. The adenovirus was spiked in the raw water and passed through the microfiltration membrane produced by Econity, a Korean membrane company, and then the viral removal rate was evaluated by real-time PCR. In the results, the amount of virus in the filtered water was decreased approximately by 5 log scale. Because coagulant treatment has been known to reduce filtering function of the membrane by inducing fouling, we also investigated whether there was any interference of coagulant. In the results, we confirmed that coagulant treatment did not show significant interference on microfiltration membrane. In this study, we found that waterborne virus can be effectively removed by membrane filtration system. In particular, here we also suggest that real-time PCR method can rapidly, sensitively and quantitatively evaluate the removal rate of virus. These results may provide a standard method to qualifying membrane filtration processes.

Analysis of cell survival genes in human gingival fibroblasts after sequential release of trichloroacetic acid and epidermal growth factor using the nano-controlled release system (나노방출제어시스템을 이용하여 trichloroacetic acid와 epidermal growth factor의 순차적 방출을 적용한 인간치은섬유아세포의 세포생존 관련 유전자 연구분석)

  • Cho, Joon Youn;Lee, Richard sungbok;Lee, Suk Won
    • Journal of Dental Rehabilitation and Applied Science
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    • v.36 no.3
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    • pp.145-157
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    • 2020
  • Purpose: This study was to determine the possible effects of trichloroacetic acid (TCA) and epidermal growth factor (EGF) through cell survival genes of the PI3K-AKT signaling pathway when applying an hydrophobically modified glycol chitosan (HGC)-based nanocontrolled release system to human gingival fibroblasts in oral soft tissue regeneration. Materials and Methods: An HGC-based nano-controlled release system was produced, followed by the loading of TCA and EGF. The group was divided into control (CON), TCA-loaded nano-controlled release system (EXP1), and the TCA- and EGF- individually loaded nano-controlled release system (EXP2). A total for 29 genes related to the PI3K-AKT signaling pathway were analyzed after 48h of culture in human gingival fibroblasts. Real-time PCR, 1- way ANOVA and multiple regression analysis were performed. Results: Cell survival genes were significantly upregulated in EXP1 and EXP2. From multiple regression analysis, ITGB1 was determined to be the most influential factor for AKT1 expression. Conclusion: The application of TCA and EGF through the HGC-based nano-controlled release system can up-regulate the cell survival pathway.

Surveillance of viable Acanthamoeba spp. and Naegleria fowleri in major water sources for tap water in Korea (한국 주요 상수원수에서의 가시아메바와 파울러자유아메바 조사)

  • Kim, Min-jeong;Lee, Gyu-Cheol;Kim, Kunwoo;Lee, Hyunji;Kim, Min Young;Seo, Dae Keun;Lee, Jeong Yeob;Cho, Young-Cheol
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.237-243
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    • 2018
  • The pathogenic free-living amoebas (FLAs), Acanthamoeba spp. and Naegleria fowleri, can cause fatal infections, including amoebic encephalitis. They are ubiquitously distributed in nature, including in diverse bodies of water. In order to survey Acanthamoeba spp. and N. fowleri in source water in Korea, we used culture-based real-time PCR to detect viable FLAs in 52 source water samples collected between July 2017 and December 2017. Acanthamoeba spp. and N. fowleri were detected in 42 samples (80.8%) and 6 samples (11.5%), respectively. Acanthamoeba spp. were detected at approximately the same frequency in all seasons, but N. fowleri was mainly detected in summer and autumn, with no N. fowleri detected in winter. These results demonstrate that these pathogenic FLAs, especially N. fowleri, which has caused deaths in the United States and China, are widely distributed in the Korean aquatic environment.

Comparison of Quantitative Relationship between Real-Time PCR and Acid Fast Bacilli Staining for Diagnosis of Pulmonary Tuberculosis (폐결핵 진단을 위한 실시간중합효소연쇄반응과 AFB 염색진단검사의 정량적 연관성 비교)

  • Jung, Taewon;Kim, Sang-Ha;Kim, Sunghyun;Choi, Jae-Sun;Kim, Young-Kwon
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.4
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    • pp.335-341
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    • 2020
  • This study investigates the association of the AFB stain with the cycle threshold (Ct) value of the Cobas TaqMan MTB test (CTM test, Roche Diagnostics, Basel, Switzerland), and it establishes the base data for semi-quantitative identification of M. tuberculosis by the Ct value. CTM test were simultaneously conducted on 8,389 specimens submitted to the Samsung Medical Center from January 2015 to December 2015, and the results were analyzed and compared retrospectively investigates the association of the AFB stain with the Ct value of the CTM test, and it establishes the base data for semi-quantitative identification of M. tuberculosis by the Ct value. The Ct values for 135 positive specimens of the CTM were inversely correlated with the AFB stain (rs=-0.545, P<0.01). When the Ct value of the CTM test and the time to positivity (TTP) of the mycobacteria cultures were verified based on the AFB stain, they were found to have a positive correlation (rs=0.136, P<0.01). The negative correlation between the CTM test and the AFB stain grade was demonstrated. The clinical significance was verified by applying these criteria to the clinical results. The semi-quantitative criteria of this study can be used to facilitate the rapid isolation of patients with active tuberculosis and infection control in the hospital.

Detection of blaKPC and blaNDM Genes from Gram-Negative Rod Bacteria Isolated from a General Hospital in Gyeongnam (경남지역 종합병원에서 분리된 그람음성막대균으로부터 blaKPC 및 blaNDM 유전자 검출)

  • Yang, Byoung Seon;Park, Ji Ae
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.1
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    • pp.49-59
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    • 2021
  • This study investigated the use of real-time PCR melting curves for the diagnosis of blaKPC and blaNDM genes among the most frequently detected carbapenemase-producing Enterobacteriaceae in Korea. As a means of addressing the shortcomings of phenotype tests and conventional PCR. The modified Hodge test confirmed positivity in 25 of 35 strains, and carbapenemase inhibition testing confirmed positivity in 14 strains by meropenem+PBA or meropenem+EDTA. PCR analysis showed amplification products in 25 strains of Klebsiella pneumoniae carbapenemases (KPC), 10 of K. pneumoniae, 5 of E. coli, 5 of A. baumannii, 4 of P. aeruginosa, and 1 of P. putida. New Delhi metallo β-lactamase (NDM) identified amplification products in 8 strains, that is, 2 K. pneumoniae, 3 E. coli, 1 P. aeruginosa, 1 E. cloacae, and 1 P. retgeri strains. Real-time PCR melting curve analysis confirmed amplification in 25 strains of KPC and 8 strains of NDM, and these results were 100% consistent with PCR results. In conclusion, our findings suggest early diagnosis of carbapenem resistant Enterobacteriaceae by real-time PCR offers a potential means of antibacterial management that can prevent and control nosocomial infection spread.

Detection of the Carbapenem Resistance Gene in Gram-negative Rod Bacteria Isolated from Clinical Specimens (임상검체에서 분리된 그람음성막대균으로부터 카바페넴 내성 유전자 검출)

  • Yang, Byoung Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.3
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    • pp.179-191
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    • 2022
  • Carbapenem-resistant Enterobacteriaceae (CRE) poses an increasing public health threat and has limited treatment options with high associated mortality. Genotypes of carbapenemase that threaten public health (blaKPC, blaNDM, blaIMP, and blaVIM) and blaOXA-48-like genes were detected by phenotypic and molecular diagnosis, and related gene distribution patterns were investigated. Phenotypic testing using the modified Hodge test confirmed positivity in all 41 strains examined, and carbapenemase inhibitory testing using meropenem+phenyl boronic acid or meropenem+EDTA confirmed positivity in 18 and 8 strains, respectively. Polymerase chain reaction revealed the presence of amplification products in 28 strains of blaKPC, 25 strains of blaNDM, 5 strains of blaIMP, 1 strain of blaVIM, and 13 blaOXA-48-like strains. In addition, 7 strains of blaKPC+blaNDM, 1 strain of blaKPC+blaIMP, 1 strain of blaNDM+blaOXA-48-like, 1 strain of blaNDM+blaVIM, 4 strains of blaKPC+blaNDM+blaIMP, and 4 strains of blaKPC+blaNDM+blaOXA-48-like were identified. Melting curve analysis using real-time PCR was wholly consistent with PCR results. The study shows genetic identification of highly specific CRE by real-time PCR could be used to provide early diagnoses and infection control, improve surveillance, and prevent the transmission of CRE.

The effects of the Reynoutria japonica on skin-barrier and moisturizing in HaCaT cells (인간유래각질형성세포에서 호장근 추출물이 피부장벽 보호능과 보습능에 미치는 영향)

  • Eun Jeong Kang;Jia Bak;Yun-Sik Choi
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.5
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    • pp.965-976
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    • 2023
  • Reynoutria japonica is a perennate plant belonging to Polygonaceae and grows wild in East Asia containing Korea. Roots of Reynoutria japonica (R. japonica), part of roots of Reynoutria japonica, has been used for anti-inflammation and antispasmodics and contains emodin as active compound. Epidermis of skin is crucial roles to defense our body against stimulants, harmful substance and prevent water loss. In this study, we examined the effect of R. japonica and emodin, its active compound, on skin-barrier and moisturizing on HaCaT cells. First, antioxidant effect of R. japonica was prominent by scavenging ABTS+ radicals. Next, we conducted real time PCR and expression of filaggrin mRNA which is crucial role in differentiation of keratinocyte increased by R. japonica and emodin dose-dependently. In addition, R. japonica and emodin significantly elevated the expression of HAS-2 mRNA which play a role in hyaluronic acid synthesis on HaCaT cells. Taken together, R. japonica containing emodin, as active compound has potential as a cosmetic material for enhancing the function of skin-barrier and moisturizing in epidermis.