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  1. Article ; Online: First study on in vitro antiviral and virucidal effects of flavonoids against feline infectious peritonitis virus at the early stage of infection

    Chanittha Triratapiban / Varanya Lueangaramkul / Nantawan Phecharat / Achiraya Pantanam / Porntippa Lekcharoensuk / Sirin Theerawatanasirikul

    Veterinary World, Vol 16, Iss 3, Pp 618-

    2023  Volume 630

    Abstract: Background and Aim: Feline infectious peritonitis (FIP), one of the most important infectious diseases in cats is caused by FIP virus (FIPV), a mutated variant of feline coronavirus. Feline infectious peritonitis has a negative impact on feline health, ... ...

    Abstract Background and Aim: Feline infectious peritonitis (FIP), one of the most important infectious diseases in cats is caused by FIP virus (FIPV), a mutated variant of feline coronavirus. Feline infectious peritonitis has a negative impact on feline health, with extremely high mortality in clinical FIP-infected cats, particularly young cats. There are no approved drugs for FIP treatment, and therapeutic possibilities for FIP treatment are limited. This study aimed to utilize nature-derived bioactive flavonoids with antiviral properties to inhibit FIPV infection in Crandell–Rees feline kidney (CRFK) cells. Materials and Methods: The cytotoxicity of 16 flavonoids was evaluated on CRFK cells using a colorimetric method (MTS) assay. Viral kinetics of FIPV at 50 tissue culture infectious dose (TCID50)/well was determined during the first 24-h post-infection (HPI). Antiviral activity was evaluated based on the replication steps of the virus life cycle, including pre-compound, attachment, penetration, post-viral entry, and virucidal assays. The antiviral efficacy of flavonoids against FIPV was determined based on positive FIPV-infected cells with the immunoperoxidase monolayer assay and viral load quantification using reverse transcription-quantitative polymerase chain reaction. Results: Two flavonoids, namely, isoginkgetin and luteolin, inhibited FIPV replication during post-viral entry in a dose-dependent manner, with 50% maximal effective concentrations = 4.77 ± 0.09 and 36.28 ± 0.03 μM, respectively. Based on viral kinetics, both flavonoids could inhibit FIPV replication at the early stage of infection at 0–6-HPI for isoginkgetin and 2–6-HPI for luteolin using a time-of-addition assay. Isoginkgetin exerted a direct virucidal effect that reduced the viral titers by 2 and 1.89 log10 TCID50/mL at 60 and 120 min, respectively. Conclusion: Isoginkgetin interfered with FIPV replication during both post-viral infection and virucidal experiments on CRFK cells, whereas luteolin inhibited the virus after infection. These results ...
    Keywords antiviral ; feline coronavirus ; feline infectious peritonitis virus ; flavonoids ; infectious disease ; Animal culture ; SF1-1100 ; Veterinary medicine ; SF600-1100
    Subject code 630 ; 570
    Language English
    Publishing date 2023-03-01T00:00:00Z
    Publisher Veterinary World
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article: Dynein light chain DYNLL1 subunit facilitates porcine circovirus type 2 intracellular transports along microtubules

    Theerawatanasirikul, Sirin / Nantawan Phecharat / Chaiwat Prawettongsopon / Wanpen Chaicumpa / Porntippa Lekcharoensuk

    Archives of virology. 2017 Mar., v. 162, no. 3

    2017  

    Abstract: Microtubule (MT) and dynein motor proteins facilitate intracytoplasmic transport of cellular proteins. Various viruses utilize microtubules and dynein for their movement from the cell periphery to the nucleus. The aim of this study was to investigate the ...

    Abstract Microtubule (MT) and dynein motor proteins facilitate intracytoplasmic transport of cellular proteins. Various viruses utilize microtubules and dynein for their movement from the cell periphery to the nucleus. The aim of this study was to investigate the intracellular transport of porcine circovirus type 2 (PCV2) via 8 kDa dynein light chain (DYNLL1, LC8) subunit along the MTs. At 20 μM, vinblastine sulfate inhibited tubulin polymerization resulting in disorganized morphology. In PCV2-infected PK-15 cells, double immunofluorescent labeling showed that the viral particles appeared at the cell periphery and gradually moved to the microtubule organization center (MTOC) at 0−12 hour post inoculation (hpi) while at 20−24 hpi they accumulated in the nucleus. Co-localization between DYNLL1 and PCV2 particles was observed clearly at 8−12 hpi. At 20−24 hpi, most aggregated tubulin had a paracrystalline appearance at the MTOC around the nucleus in vinblastine-treated, PCV2-infected PK-15 cells. Between 12 and 24 hpi, PCV2 particles were still bound to DYNLL1 before they were translocated to the nucleus in both treatments, indicating that vinblastine sulfate had no effect on the protein-protein co-localization. The DYNLL1 binding motif, LRLQT, was found near the C-terminus of PCV2 capsid protein (Cap). Molecular docking analysis confirmed the specific interaction between these residues and the cargo binding site on DYNLL1. Our study clearly demonstrated that dynein, in particular DYNLL1, mediated PCV2 intracellular trafficking. The results could explain, at least in part, the viral transport mechanism by DYNLL1 via MT during PCV2 infection.
    Keywords Porcine circovirus-2 ; binding sites ; coat proteins ; dynein ATPase ; fluorescent labeling ; microtubules ; molecular models ; molecular motor proteins ; physiological transport ; polymerization ; sulfates ; tubulin ; vinblastine ; virion ; viruses
    Language English
    Dates of publication 2017-03
    Size p. 677-686.
    Publishing place Springer Vienna
    Document type Article
    ZDB-ID 7491-3
    ISSN 1432-8798 ; 0304-8608
    ISSN (online) 1432-8798
    ISSN 0304-8608
    DOI 10.1007/s00705-016-3140-0
    Database NAL-Catalogue (AGRICOLA)

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  3. Article: Cloning and production of nucleoprotein of swine influenza virus in E.coli

    Areerat Pangpeng(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) E-mail:areejum@hotmail.com / Nantawan Phecharat(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) / Porntippa Lekcharoensuk(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology)

    Abstract: Influenza A virus is a causative agent of swine influenza which is an acute respiratory disease affecting pigs at all ages leading to considerable economic losses of swine industries. Monitoring and surveillance of swine influenza virus (SIV) infection ... ...

    Abstract Influenza A virus is a causative agent of swine influenza which is an acute respiratory disease affecting pigs at all ages leading to considerable economic losses of swine industries. Monitoring and surveillance of swine influenza virus (SIV) infection and herd immunity are essential for disease prevention and control. However, it requires an effective diagnostic tool such as ELISA. Nucleoprotein (NP) is a major group specific and conserved protein suitable to be use as antigen for the detection of all subtypes of influenza A viruses. The objective of this study was to production and purification of the recombinant NP protein. Total RNA was extracted from swine influenza virus (A/swine/IA/1930) suspension. cDNA was synthesis from the total RNA and NP gene was amplified using NP specific primers which designed to contain KpnI and SalI at 5 ends of the forward and reverse primers, respectively. The NP gene was ligated to the vector, pQE80L, prior to be transformed into E.coli strain DH5 alpha. In E.coli. The expressed protein reached the highest yield when the expression was induced using 0.2 mM IPTG at 37 deg C for 24 hours. Majority of the protein was insoluble and accumulated as inclusion bodies. The insoluble NP protein was dissolved in 8 M Urea pH 8.0 and purified using a nickel-charged resin-affinity column. The protein was then neutralized using stepwise dilution method. The recombinant protein had a molecular weight approximately 56 kDa and reacted strongly to convalescent serum from a pig infected with SIV. The results showed that the recombinant NP protein was successfully produced. It is immunogenic and ready for further diagnostic application
    Keywords Swine ; Viruses ; Animal diseases ; Viroses ; Influenzavirus ; Molecular cloning ; Genetic engineering ; Animal production ; Nucleoproteins
    Language Thai
    Document type Article
    Database AGRIS - International Information System for the Agricultural Sciences and Technology

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  4. Article: Development of quantitative competitive PCR for the detection of porcine circovirus type 2 DNA in pigs affected with PMWS

    Nattarat Thangthamniyom(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) E-mail:j2gole@yahoo.com / Porntippa Lekcharoensuk(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) / Tippawan Jantafong(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology) / Nantawan Phecharat(Kasetsart University, Bangkok (Thailand). Faculty of Veterinary Medicine. Department of Microbiology and Immunology)

    Abstract: Porcine circovirus type 2 (PCV2) is an essential infectious agent of postweaning multisystemic wasting syndrome (PMWS). In this study, a quantitative competitive PCR (QC-PCR) assay was developed for monitoring PCV DNA in fecal swab from pigs in PMWS- ... ...

    Abstract Porcine circovirus type 2 (PCV2) is an essential infectious agent of postweaning multisystemic wasting syndrome (PMWS). In this study, a quantitative competitive PCR (QC-PCR) assay was developed for monitoring PCV DNA in fecal swab from pigs in PMWS-negative and -affected farms. The QC-PCR was based on competitive co-amplification of a 345 bp of the PCV type 2 with a known concentration of the competitor DNA, which produced a 513 bp fragment. The result demonstrated that the PCV2 DNA content obtained form the samples from the PMWS-affected farms higher than 1 pg/ul whereas those from the PMWS-negative farms was lower than 1 fg/ul with the exception of 10pg/ul of the sample F39. According to the results, the amount of PCV2 DNA load from clinical PMWS free and clinical PMWS affected farms were different significantly and this indicating that QC-PCR may be a useful tool for predicting of the occurrence of PMWS in pig farms.
    Keywords Swine ; Animal diseases ; Morbidity ; PCR ; DNA
    Language Thai
    Document type Article
    Database AGRIS - International Information System for the Agricultural Sciences and Technology

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