• Title/Summary/Keyword: DNA preparation

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Comparison of Loop-mediated Isothermal Amplification and Korea Standard Food Codex (KFSC) Method for Detection of Salmonella Typhimurium, Listeria monocytogenes Artificially Inoculated in Yuk-hwe and Yuk-sashimi (육회와 육사시미에 접종된 Salmonella Typhimurium와 Listeria monocytogenes 검출을 위한 Loop-mediated isothermal amplification와 식품공전의 배지 시험법, real-time PCR의 검출 성능 비교)

  • Gwak, Seung-Hae;Lee, So-Young;Kim, Jin-Hee;Oh, Se-Wook
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.277-282
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    • 2019
  • The object of this study is to compare the performance of the 3M Molecular Detection Assay 2 (3M MDA 2) and the Korea Standard Food Codex (KSFC) Method (i.e., isolation media and real-time PCR) in detecting Salmonella Typhimurium and Listeria monocytogenes in traditional Korean foods. Yuk-hwe and Yuk-sashimi (types of raw beef dishes) were artificially inoculated with $10^0-10^4CFU/25g$ of L. monocytogenes and S. Typhimurium. Citrobacter freundii and Listeria innocua were used as competitive microflora. After enrichment, the samples were analyzed using 3M MDA 2 and real-time PCR. All samples inoculated at concentrations of $10^0-10^4CFU/25g$ without competitive microflora were positive for S. Typhimurium and L. monocytogenes, as detected by 3M MDA 2 and Korea Standard Food Codex (KFSC) Method. In addition, part of the samples were positive for the presence of C. freundii and L. innocua. The 3M MDA 2 - Salmonella and Korea Standard Food Codex (KFSC) Method showed similar detection performances in Yuk-hwe and Yuk-sashimi. The 3M MDA 2 method for Salmonella and Listeria, which is a LAMP-based technology, can be used for rapid detection of S. Typhimurium and L. monocytogenes in raw beef. LAMP bioluminescence assays provide results on the subsequent day and are simple to use compared with the Korea Standard Food Codex (KFSC) Method, particularly in terms of DNA preparation.

Inhibitory Effect of Hot-Water Extract of Paeonia japonica on Oxidative Stress and Identification of Its Active Components (백작약 열수추출물의 산화적 스트레스 억제효과 및 유효성분 동정)

  • Jeong, Ill-Yun;Lee, Joo-Sang;Oh, Heon;Jung, U-Hee;Park, Hae-Ran;Jo, Sung-Kee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.5
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    • pp.739-744
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    • 2003
  • This study was carried out to investigate the antioxidative activity and to identify the active components of hot-water extract of Paeoniajaponica (PJ), which was a main ingredient of a herb mixture preparation recently established as a potent candidate of radioprotector in our laboratory. The water extract was fractionated with CHCl$_3$, EtOAc and n-BuOH. The extract and its fractions showed very low activity in hydroxyl radical scavenging test. In lipid peroxidation test, the extract, EtOAc and water fractions showed moderate inhibition with the ratio above 50%. In DPPH radical scavenging test, the extract, EtOAc and water fraction showed high activity with the ratio above 80%, especially. EtOAc fraction scavenged the radicals as much as synthetic antioxidant (BHA), even at low concentration. It is suggested that mai or partition for antioxidative activity of Paeonia japonica was EtOAc fraction. Subsequently, two active compounds (PJE021-1 and JE024-1) from EtOAc fraction were isolated by using MCI gel and silica gel column chromatography The two compounds inhibited remarkedly the $H_2O$$_2$-induced DNA damage in human peripheral blood lymphocytes, measured by single-cell gel electrophoresis (SCGE). PJE021-1 protected the cells to almost negative control level, dose-dependently. PJE024-1 exhibited a potent inhibition with the ratio of 71% at even low concentration (0.5 $\mu\textrm{g}$/$m\ell$). Finally, their chemical structures were identified as gallic acid (PJE021-1) and (+)-catechin (PJE024-1), respectively, on the basis of the speculation of spectral and physical data.

Multiplication of Infectious Flacherie and Densonucleosis Viruses in the Silkworm, Bombyx mori (가잠의 전염성 연화병 및 농핵병 바이러스 증식에 관한 연구)

  • 김근영;강석권
    • Journal of Sericultural and Entomological Science
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    • v.25 no.2
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    • pp.1-31
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    • 1984
  • Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

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