• Title/Summary/Keyword: MDCK cell

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Screening of Antiviral Medicinal Plants against Avian Influenza Virus H1N1 for Food Safety

  • Lee, Jang-Hyun;Van, Nguyen Dinh; Ma, Jin-Yeul;Kim, Young-Bong;Kim, Soo-Ki;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.30 no.2
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    • pp.345-350
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    • 2010
  • Various extracts from 30 medicinal plants were evaluated for their antiviral activity against influenza virus A/Puerto Rico/8/34 (H1N1) and cytotoxicity in MDCK cell culture. The plant material (30 g) was extracted with methanol (300 mL) at room temperature for 24 h, after which the methanolic extracts were filtered, evaporated, and subsequently lyophilized. Evaluation of the potential antiviral activity was conducted by a viral replication inhibition test. Among these medicinal plants, Tussilago farfara, Brassica juncea, Prunus armeniaca, Astragalus membranaceus, Patrinia villosa, and Citrus unshiu showed marked antiviral activity against influenza virus A/H1N1 at concentrations ranging from 0.15625 mg/mL to 1.25 mg/mL, 0.3125 mg/mL to 10 mg/mL, 5 mg/mL to 10 mg/mL, 0.625 mg/mL to 10 mg/mL, 0.625 mg/mL to 10 mg/mL, and 0.3125 mg/mL to 5 mg/mL, respectively. The extracts of Tussilago farfara showed cytotoxicity at concentrations greater than 2.5 mg/mL, whereas the other five main extracts showed no cytotoxicity at concentrations of 10 mg/mL. Taken together, the present results indicated that methanolic extracts of the six main plants might be useful for the treatment of influenza virus H1N1.

Elucidation of the Inhibitory Mechanisms of Nipponoparmelia laevior Lichen Extract against Influenza A (H1N1) Virus through Proteomic Analyses

  • Cuong, Tran Van;Cho, Se-Young;Kwon, Joseph;Kim, Duwoon
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1155-1164
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    • 2019
  • Lichens contain diverse bioactive secondary metabolites with various chemical and biological properties, which have been widely studied. However, details of the inhibitory mechanisms of their secondary metabolites against influenza A virus (IAV) have not been documented. Here, we investigated the antiviral effect of lichen extracts, obtained from South Korea, against IAV in MDCK cells. Of the lichens tested, Nipponoparmelia laevior (LC24) exhibited the most potent inhibitory effect against IAV infection. LC24 extract significantly increased cell viability, and reduced apoptosis in IAV-infected cells. The LC24 extract also markedly reduced (~ 3.2 log-fold) IAV mRNA expression after 48 h of infection. To understand the antiviral mechanism of LC24 against IAV, proteomic (UPLC-$HDMS^E$) analysis was performed to compare proteome modulation in IAV-infected (V) vs. mock (M) and LC24+IAV (LCV) vs. V cells. Based on Ingenuity Pathway Analysis (IPA), LC24 inhibited IAV infection by modulating several antiviral-related genes and proteins (HSPA4, HSPA5, HSPA8, ANXA1, ANXA2, $HIF-1{\alpha}$, AKT1, MX1, HNRNPH1, HNRNPDL, PDIA3, and VCP) via different signaling pathways, including $HIF-1{\alpha}$ signaling, unfolded protein response, and interferon signaling. These molecules were identified as the specific biomarkers for controlling IAV in vitro and further confirmation of their potential against IAV in vivo is required. Our findings provide a platform for further studies on the application of lichen extracts against IAV.

Expression, Purification and Antiserum Production of the Avian Influenza H9N2 Virus HA and NA Proteins (Avian Influenza H9N2 Virus의 HA와 NA 단백질 발현, 정제 및 항혈청 생산)

  • Lee, Hyun-Ji;Song, Byung-Hak;Kim, Jeong-Min;Yun, Sang-Im;Kim, Jin-Kyoung;Kang, Young-Sik;Koo, Yong-Bum;Jeon, Ik-Soo;Byun, Sung-June;Lee, Youn-Jeong;Kwon, Jun-Hun;Park, Jong-Hyeon;Joo, Yi-Seok;Lee, Young-Min
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.178-185
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    • 2008
  • Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.

Viral Patterns and Clincal Analysis of Acute Respiratory Tract Infections of Children in Korea(September, 1998~August, 2002) (바이러스에 의한 소아 급성 하기도 감염의 유행 및 임상양상(1998년 9월~2002년 8월))

  • Lee, Su-Jin;Park, Eun-Young;Oh, Phil-Soo;Lee, Kon-Hee;Kim, Kwang-Nam;Lee, Kyu-Man
    • Pediatric Infection and Vaccine
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    • v.10 no.1
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    • pp.102-113
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    • 2003
  • Purpose : Acute respiratory tract infections are the most common illnesses in children. The great majority of these infections involving lower respiratory tracts infections(LRTI) are caused by respiratory viruses such as respiratory syncytial virus(RSV), parainfluenza virus (PIV), influenza virus(INF), adenovirus(ADV), etc. Our purpose of this study was to examine the seasonal occurrence, clinical and laboratory features of each viral infections. Methods : Nasopharyngeal aspirates(NPA) were collected from 913 hospitalized children on the first day of admission. They were all admitted to Hangang Sacred Heart hospital and diagnosed as LRTIs. The study period was from September 1998 to August 2002. Specimens were inoculated onto HEp-2 cell, LLC-MK2 cell and MDCK cell. Viruses were detected by immunofluorescent method performed at day 10 postinoculation. Respiratory viruses were detected in 251(27%) cases. Medical records of 208 cases diagnosed as acute LRTIs were all taken reviewed and analyzed. Results : The identified pathogens were RSV 122 cases(58%), PIV 30 cases(14%), INFA 20 cases(10%), INFB 21 cases(11%) and ADV 15 cases(7%) during the study period. Outbreaks of RSV infections occurred every year but mostly on December, 1998 and November, 1999. LRTIs by PIV and ADV occurred during all seasons of the year. INFA was isolated moslty on March, 1999 and January, 2000. INFB infections occurred largely on April, 2002. The clinical diagnoses were bronchiolitis 88 cases(43%), pneumonia 87 cases(42%), croup 19 cases(9%) and tracheobronchitis 14 cases(6%). Common symptoms and signs were cough, rhinorrhea, sputum and fever. WBC counts in peripheral blood showed normal findings in 123 cases(59%). Each viral infections also showed some differences in auscultatory findings and chest X-ray findings. Conclusion : We learned that viruses are one of the major and important etiologic agents of acute LRTIs of children in Korea. However, we still need to find out more about its characteristic clinical features and continue studying on their seasonal occurrence to focus on their management and also prevention.

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