• 제목/요약/키워드: Real-Time Polymerase Chain Reaction

검색결과 795건 처리시간 0.031초

Gene expression profiling after ochratoxin A treatment in small intestinal epithelial cells from pigs

  • Jung Woong, Yoon;Sang In, Lee
    • Journal of Animal Science and Technology
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    • 제64권5호
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    • pp.842-853
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    • 2022
  • Ochratoxin A (OTA) is a well-known mycotoxin that causes disease through the ingestion of contaminated food or feed, for example, in the porcine industry. The intestinal epithelium acts as the first barrier against food contamination. We conducted a study on the exposure of the porcine intestinal epithelium to OTA. We used the intestinal porcine epithelial cell line IPEC-J2 as an in vitro model to evaluate the altered molecular mechanisms following OTA exposure. Gene expression profiling revealed that OTA upregulated 782 genes and downregulated 896, totalling 1678 differentially expressed genes. Furthermore, immunofluorescence, quantitative real-time polymerase chain reaction, and western blotting confirmed that OTA damages the tight junction protein ZO-1. Moreover, OTA activated the expression of inflammatory genes (IL-6, IL-8, IL-10, NF-kB, TLR4, and TNF-α). In summary, this study confirmed that OTA alters various molecular mechanisms and has several adverse effects on IPEC-J2 cells.

Successful Management of Eosinophilic Bronchopneumopathy in a Dog

  • Kim, Sol;Seo, Kyoungwon;Song, Kunho
    • 한국임상수의학회지
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    • 제38권6호
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    • pp.269-273
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    • 2021
  • A dog was presented with a cough, dyspnea, nasal discharge, gagging, and exercise intolerance. The dog showed leukocytosis, peripheral eosinophilia, and an increase C-reactive protein. The radiographic findings noted bronchointerstitial infiltration, intrathoracic lymphadenopathy, and soft tissue opacity mass. Computed tomography findings showed thickening of the bronchus and bronchiole. Also, peri-bronchial consolidation and generalized intrathoracic lymphadenopathy was present. On blind bronchoalveolar lavage and pulmonary cytology, there were significantly increased eosinophils. Canine pulmonary respiratory pathogens from a real-time polymerase chain reaction analysis was negative. Consequently, the dog was diagnosed with eosinophilic bronchopneumopathy. Clinical signs improved significantly within a few days after treatment with an oral corticosteroid.

Anti-Inflammatory Effect of Carex scabrifolia Steud. Extract in RAW264.7 Cells

  • Joong Hyun Shim
    • 한국미생물·생명공학회지
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    • 제50권3호
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    • pp.354-360
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    • 2022
  • This research was designed to evaluate the possible anti-inflammatory effects of Carex scabrifolia Steud. extract using RAW264.7 cells. The assessments of these effects were based on cell viability assay, mRNA expression levels of interleukin-1 alpha (IL-1α), interleukin-1 beta (IL-1β), IL-6, tumor necrosis factor alpha (TNFα), and levels of nitric oxide (NO)/prostaglandin E2 (PGE2) production. Quantitative real-time polymerase chain reaction showed that treatment with C. scabrifolia Steud. extract decreased the mRNA levels of iNOS, COX2, IL-1α, IL-1β, IL-6, and TNFα. Furthermore, from the production levels of PGE2/NO, it can be inferred that C. scabrifolia Steud. extract exhibited anti-inflammatory properties. These results suggest that C. scabrifolia Steud. extract contains anti-inflammatory compound(s), and consequently, that it may have applications as a potent cosmeceutical material.

공기 중 박테리아 포집을 위한 습식 사이클론의 CFD 해석을 이용한 설계 및 성능 평가 (Design and Performance Evaluation using Computational Fluid Dynamics (CFD) Analysis of Wetcyclones for the Collection of Airborne Bacteria)

  • 고현식;박정우;정지우;황정호
    • 한국입자에어로졸학회지
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    • 제19권3호
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    • pp.77-87
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    • 2023
  • We present the development of a wetcyclone sampler designed for the sampling of airborne bacteria. The wetcyclone design involves a combination of two traditional cyclone shapes and computational fluid dynamics (CFD) analysis to validate its effectiveness in terms of pressure drop and collection efficiency. The wetcyclone exhibits a collection efficiency of over 90% for bacteria, specifically targeting Staphylococcus aureus. Additionally, the wetcyclone enables continuous bioaerosol sampling using a liquid medium (deionized water), demonstrating a concentration ratio exceeding >105 and a stable microbial recovery rate of 81.9%. The application of real-time quantitative polymerase chain reaction (qPCR) and the colony counting method ensures precise measurement of the concentration ratio and microbial recovery rate.

2019 Novel Coronavirus (COVID-19) Pneumonia: Serial Computed Tomography Findings

  • Jiangping Wei;Huaxiang Xu;Jingliang Xiong;Qinglin Shen;Bing Fan;Chenglong Ye;Wentao Dong;Fangfang Hu
    • Korean Journal of Radiology
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    • 제21권4호
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    • pp.501-504
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    • 2020
  • From December 2019, Coronavirus disease 2019 (COVID-19) pneumonia (formerly known as the 2019 novel Coronavirus [2019-nCoV]) broke out in Wuhan, China. In this study, we present serial CT findings in a 40-year-old female patient with COVID-19 pneumonia who presented with the symptoms of fever, chest tightness, and fatigue. She was diagnosed with COVID-19 infection confirmed by real-time reverse-transcriptase-polymerase chain reaction. CT showed rapidly progressing peripheral consolidations and ground-glass opacities in both lungs. After treatment, the lesions were shown to be almost absorbed leaving the fibrous lesions.

Development of a ladder-shape melting temperature isothermal amplification (LMTIA) assay for detection of African swine fever virus (ASFV)

  • Wang, Yongzhen;Wang, Borui;Xu, Dandan;Zhang, Meng;Zhang, Xiaohua;Wang, Deguo
    • Journal of Veterinary Science
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    • 제23권4호
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    • pp.51.1-51.10
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    • 2022
  • Background: Due to the unavailability of an effective vaccine or antiviral drug against the African swine fever virus (ASFV), rapid diagnosis methods are needed to prevent highly contagious African swine fever. Objectives: The objective of this study was to establish the ladder-shape melting temperature isothermal amplification (LMTIA) assay for the detection of ASFV. Methods: LMTIA primers were designed with the p72 gene of ASFV as the target, and plasmid pUC57 was used to clone the gene. The LMTIA reaction system was optimized with the plasmid as the positive control, and the performance of the LMTIA assay was compared with that of the commercial real-time polymerase chain reaction (PCR) kit in terms of sensitivity and detection rate using 200 serum samples. Results: Our results showed that the LMTIA assay could detect the 104 dilution of DNA extracted from the positive reference serum sample, which was the same as that of the commercial real-time PCR kit. The coincidence rate between the two assays was 100%. Conclusions: The LMTIA assay had high sensitivity, good detection, and simple operation. Thus, it is suitable for facilitating preliminary and cost-effective surveillance for the prevention and control of ASFV.

Prevalence of feline calicivirus in Korean cats determined by an improved real-time RT-PCR assay

  • Ji-Su Baek;Jong-Min Kim;Hye-Ryung Kim;Yeun-Kyung Shin;Oh-Kyu Kwon;Hae-Eun Kang;Choi-Kyu Park
    • 한국동물위생학회지
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    • 제46권2호
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    • pp.123-135
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    • 2023
  • Feline calicivirus (FCV) is considered the main viral pathogen of feline upper respiratory tract disease (URTD). The frequent mutations of field FCV strains result in the poor diagnostic sensitivity of previously developed molecular diagnostic assays. In this study, a more sensitive real-time reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed for broad detection of currently circulating FCVs and comparatively evaluated the diagnostic performance with previously developed qRT-PCR assay using clinical samples collected from Korean cat populations. The developed qRT-PCR assay specifically amplified the FCV p30 gene with a detection limit of below 10 copies/reaction. The assay showed high repeatability and reproducibility, with coefficients of intra-assay and inter-assay variation of less than 2%. Based on the clinical evaluation using 94 clinical samples obtained from URTD-suspected cats, the detection rate of FCV by the developed qRT-PCR assay was 47.9%, which was higher than that of the previous qRT-PCR assay (43.6%). The prevalence of FCV determined by the new qRT-PCR assay in this study was much higher than those of previous Korean studies determined by conventional RT-PCR assays. Due to the high sensitivity, specificity, and accuracy, the new qRT-PCR assay developed in this study will serve as a promising tool for etiological and epidemiological studies of FCV circulating in Korea. Furthermore, the prevalence data obtained in this study will contribute to expanding knowledge about the epidemiology of FCV in Korea.

Identification and Analysis of the Chloroplast rpoC1 Gene Differentially Expressed in Wild Ginseng

  • Lee, Kwang-Ho;Kwon, Ki-Rok;Kang, Won-Mo;Jeon, Eun-Mi;Jang, Jun-Hyeog
    • 대한약침학회지
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    • 제15권2호
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    • pp.20-23
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    • 2012
  • Panax ginseng is a well-known herbal medicine in traditional Asian medicine, and wild ginseng is widely accepted to be more active than cultivated ginseng in chemoprevention. However, little has actually been reported on the difference between wild ginseng and cultivated ginseng. Thus, to identify and analyze those differences, we used suppressive subtraction hybridization (SSH) sequences with microarrays, realtime polymerase chain reaction (PCR), and reverse transcription PCRs (RT-PCRs). One of the clones isolated in this research was the chloroplast rpoC1 gene, a ${\beta}$subunit of RNA polymerase. Real-time RT-PCR results showed that the expression of the rpoC1 gene was significantly upregulated in wild ginseng as compared to cultivated ginseng, so, we conclude that the rpoC1 gene may be one of the important markers of wild ginseng.

Quantitative Real-Time RT-PCR of ITGA7, SVEP1, TNS1, LPHN3, SEMA3G, KLB and MMP13 mRNA Expression in Breast Cancer

  • Kotepui, Manas;Thawornkuno, Charin;Chavalitshewinkoon-Petmitr, Porntip;Punyarit, Phaibul;Petmitr, Songsak
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권11호
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    • pp.5879-5882
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    • 2012
  • Breast cancer is the leading cause of cancer deaths among women worldwide, including Thailand. In the present study, the differential mRNA expression of SVEP1, LPHN3, KLB, ITGA7, SEMA3G, TNS1 and MMP13 genes was examined in breast cancer using quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR). Among these genes, increased LPHN3 and MMP13 mRNA expression levels correlated with axillary-node metastasis (P=0.02). Multiple logistic regression analysis revealed that LPHN3 and MMP13 mRNA expression is significantly associated with axillary node status in breast cancer (P=0.04).

Application of the melanocortin 1 receptor (MC1R) gene for discrimination of Hanwoo from Holstein beef using real-time polymerase chain reaction (PCR)

  • Ra, Do-Kyung;Lee, Sung-Mo;Park, Eun-Jeong;Lee, Jung-Goo
    • 한국동물위생학회지
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    • 제30권4호
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    • pp.557-562
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    • 2007
  • This study was carried out to discriminate Hanwoo from the milking and hybrid cattle by detection of MC1R gene related to bovine hair color. One hundred sixty six samples were collected from the abattoir (n = 106) and local market (n = 60). The beef from abattoir were originated from Hanwoo (n=27), Holstein (n=29), Hybrid (n=45) and imported cattle (n=5), respectively. The beef from market consisted of Hanwoo (n=36), Holstein (n=7) and imported ones (n=17). Commercialized screening kit (Kogenebiotec, Korea) was used for MC1R gene analysis. As a result, Hanwoo was discriminated from Holstein. However, 9 of 45 hybrid and 11 of 22 imported beef samples were indistinguishable from Hanwoo. It could be explained by second generation of crossing of Hanwoo with Holstein or the cattle with silver or yellow hair. This results suggest that additional tests as well as MC1R gene detection be needed to confirm Hanwoo beef among cattle beef.