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  1. Article ; Online: Proteolytic enzymes in embryonated chicken eggs sustain the replication of egg-grown low-pathogenicity avian influenza viruses in cells in the absence of exogenous proteases.

    Kandeil, Ahmed / Bagato, Ola / Zaraket, Hassan / Debeauchamp, Jennifer / Krauss, Scott / El-Shesheny, Rabeh / Webby, Richard J / Ali, Mohamed A / Kayali, Ghazi

    Journal of virological methods

    2014  Volume 202, Page(s) 28–33

    Abstract: Low pathogenic influenza viruses grow readily in embryonated chicken eggs but require the addition of exogenous proteases to grow in MDCK cell culture. In this study, we found that the influenza viruses propagated previously in eggs, can grow for up to ... ...

    Abstract Low pathogenic influenza viruses grow readily in embryonated chicken eggs but require the addition of exogenous proteases to grow in MDCK cell culture. In this study, we found that the influenza viruses propagated previously in eggs, can grow for up to two passages in cell culture without the addition of exogenous proteolytic enzymes. These results indicate that the reason for virus propagation in cells during the first two passages may be due to proteases from egg allantoic fluid carried over from egg culture. The ability of influenza viruses to grow in cells in the absence of trypsin is currently considered as a hallmark of highly pathogenic influenza viruses. Our data indicate that differentiating between high and low pathogenicity using cell culture only is not appropriate and other indicators such as sequence analysis and in vitro pathogenicity index should be performed.
    MeSH term(s) Animals ; Cell Line ; Chickens ; Dogs ; Influenza A virus/classification ; Influenza A virus/growth & development ; Influenza A virus/pathogenicity ; Influenza A virus/physiology ; Influenza in Birds/virology ; Peptide Hydrolases/metabolism ; Virulence ; Virus Cultivation/methods ; Virus Replication ; Zygote/enzymology ; Zygote/virology
    Chemical Substances Peptide Hydrolases (EC 3.4.-)
    Language English
    Publishing date 2014-03-10
    Publishing country Netherlands
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 8013-5
    ISSN 1879-0984 ; 0166-0934
    ISSN (online) 1879-0984
    ISSN 0166-0934
    DOI 10.1016/j.jviromet.2014.02.023
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Proteolytic enzymes in embryonated chicken eggs sustain the replication of egg-grown low-pathogenicity avian influenza viruses in cells in the absence of exogenous proteases

    Kandeil, Ahmed / Bagato, Ola / Zaraket, Hassan / Debeauchamp, Jennifer / Krauss, Scott / El-Shesheny, Rabeh / Webby, Richard J / Ali, Mohamed A / Kayali, Ghazi

    Elsevier B.V. Journal of virological methods. 2014 June 15, v. 202

    2014  

    Abstract: Low pathogenic influenza viruses grow readily in embryonated chicken eggs but require the addition of exogenous proteases to grow in MDCK cell culture. In this study, we found that the influenza viruses propagated previously in eggs, can grow for up to ... ...

    Abstract Low pathogenic influenza viruses grow readily in embryonated chicken eggs but require the addition of exogenous proteases to grow in MDCK cell culture. In this study, we found that the influenza viruses propagated previously in eggs, can grow for up to two passages in cell culture without the addition of exogenous proteolytic enzymes. These results indicate that the reason for virus propagation in cells during the first two passages may be due to proteases from egg allantoic fluid carried over from egg culture. The ability of influenza viruses to grow in cells in the absence of trypsin is currently considered as a hallmark of highly pathogenic influenza viruses. Our data indicate that differentiating between high and low pathogenicity using cell culture only is not appropriate and other indicators such as sequence analysis and in vitro pathogenicity index should be performed.
    Keywords Influenza A virus ; allantoic fluid ; cell culture ; chicken eggs ; eggs ; pathogenicity ; sequence analysis ; trypsin ; viruses
    Language English
    Dates of publication 2014-0615
    Size p. 28-33.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 8013-5
    ISSN 1879-0984 ; 0166-0934
    ISSN (online) 1879-0984
    ISSN 0166-0934
    DOI 10.1016/j.jviromet.2014.02.023
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  3. Article ; Online: Impact of Adjuvants on the Immunogenicity and Efficacy of Split-Virion H7N9 Vaccine in Ferrets.

    Wong, Sook-San / Kaplan, Bryan / Zanin, Mark / Debeauchamp, Jennifer / Kercher, Lisa / Crumpton, Jeri-Carol / Seiler, Patrick / Sun, Yilun / Tang, Li / Krauss, Scott / Webster, Robert / Webby, Richard J

    The Journal of infectious diseases

    2015  Volume 212, Issue 4, Page(s) 542–551

    Abstract: Background: An effective vaccine is urgently needed against the H7N9 avian influenza virus. We evaluated the immunogenicity and protective efficacy of a split-virion H7N9 vaccine with or without the oil-in-water adjuvants in ferrets.: Methods: ... ...

    Abstract Background: An effective vaccine is urgently needed against the H7N9 avian influenza virus. We evaluated the immunogenicity and protective efficacy of a split-virion H7N9 vaccine with or without the oil-in-water adjuvants in ferrets.
    Methods: Ferrets were vaccinated with 2 doses of unadjuvanted, MF59 or AS03-adjuvanted A/Shanghai/2/2013 (H7N9) vaccine, and the induction of antibodies to hemagglutinin (HA) or neuraminidase proteins was evaluated. Ferrets were then challenged with wild-type H7N9 virus to assess the vaccine's protective efficacy. The vaccine composition and integrity was also evaluated in vitro.
    Results: Adjuvanted vaccines stimulated robust serum antibody titers against HA and neuraminidase compared with the unadjuvanted vaccines. Although there was a difference in adjuvanticity between AS03 and MF59 at a lower dose (3.75 µg of HA), both adjuvants induced comparable antibody responses after 2 doses of 15 µg. On challenge, ferrets that received adjuvanted vaccines showed lower viral burden than the control or unadjuvanted vaccine group. In vitro examinations revealed that the vaccine contained visible split-virus particles and retained the native conformation of HA recognizable by polyclonal and monoclonal antibodies.
    Conclusions: The adjuvanted H7N9 vaccines demonstrated superior immunogenicity and protective efficacy against H7N9 infection in ferrets and hold potential as a vaccination regimen.
    MeSH term(s) Adjuvants, Immunologic/pharmacology ; Animals ; Antibodies, Viral/biosynthesis ; Antibodies, Viral/blood ; Cross Reactions ; Dose-Response Relationship, Immunologic ; Drug Combinations ; Ferrets ; Influenza A Virus, H7N9 Subtype/immunology ; Influenza Vaccines/administration & dosage ; Influenza Vaccines/immunology ; Male ; Polysorbates/administration & dosage ; Polysorbates/pharmacology ; Specific Pathogen-Free Organisms ; Squalene/administration & dosage ; Squalene/pharmacology ; alpha-Tocopherol/administration & dosage ; alpha-Tocopherol/pharmacology
    Chemical Substances Adjuvants, Immunologic ; Antibodies, Viral ; Drug Combinations ; Influenza Vaccines ; MF59 oil emulsion ; Polysorbates ; Squalene (7QWM220FJH) ; AS03 adjuvant (A7YT618XBV) ; alpha-Tocopherol (H4N855PNZ1)
    Language English
    Publishing date 2015-02-23
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 3019-3
    ISSN 1537-6613 ; 0022-1899
    ISSN (online) 1537-6613
    ISSN 0022-1899
    DOI 10.1093/infdis/jiv099
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.MO 445: Show issues

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  4. Article ; Online: Evidence of infection with H4 and H11 avian influenza viruses among Lebanese chicken growers.

    Kayali, Ghazi / Barbour, Elie / Dbaibo, Ghassan / Tabet, Carelle / Saade, Maya / Shaib, Houssam A / Debeauchamp, Jennifer / Webby, Richard J

    PloS one

    2011  Volume 6, Issue 10, Page(s) e26818

    Abstract: Human infections with H5, H7, and H9 avian influenza viruses are well documented. Exposure to poultry is the most important risk factor for humans becoming infected with these viruses. Data on human infection with other low pathogenicity avian influenza ... ...

    Abstract Human infections with H5, H7, and H9 avian influenza viruses are well documented. Exposure to poultry is the most important risk factor for humans becoming infected with these viruses. Data on human infection with other low pathogenicity avian influenza viruses is sparse but suggests that such infections may occur. Lebanon is a Mediterranean country lying under two major migratory birds flyways and is home to many wild and domestic bird species. Previous reports from this country demonstrated that low pathogenicity avian influenza viruses are in circulation but highly pathogenic H5N1 viruses were not reported. In order to study the extent of human infection with avian influenza viruses in Lebanon, we carried out a seroprevalence cross-sectional study into which 200 poultry-exposed individuals and 50 non-exposed controls were enrolled. We obtained their sera and tested it for the presence of antibodies against avian influenza viruses types H4 through H16 and used a questionnaire to collect exposure data. Our microneutralization assay results suggested that backyard poultry growers may have been previously infected with H4 and H11 avian influenza viruses. We confirmed these results by using a horse red blood cells hemagglutination inhibition assay. Our data also showed that farmers with antibodies against each virus type clustered in a small geographic area suggesting that unrecognized outbreaks among birds may have led to these human infections. In conclusion, this study suggests that occupational exposure to chicken is a risk factor for infection with avian influenza especially among backyard growers and that H4 and H11 influenza viruses may possess the ability to cross the species barrier to infect humans.
    MeSH term(s) Adult ; Agriculture ; Animal Husbandry ; Animals ; Case-Control Studies ; Chickens ; Female ; Humans ; Influenza A virus ; Influenza in Birds/transmission ; Influenza in Birds/virology ; Influenza, Human/transmission ; Influenza, Human/virology ; Lebanon ; Male ; Middle Aged ; Occupational Exposure
    Language English
    Publishing date 2011-10-28
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2267670-3
    ISSN 1932-6203 ; 1932-6203
    ISSN (online) 1932-6203
    ISSN 1932-6203
    DOI 10.1371/journal.pone.0026818
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  5. Article ; Online: A single dose of whole inactivated H7N9 influenza vaccine confers protection from severe disease but not infection in ferrets.

    Wong, Sook-San / Jeevan, Trushar / Kercher, Lisa / Yoon, Sun-Woo / Petkova, Atanaska-Marinova / Crumpton, Jeri-Carol / Franks, John / Debeauchamp, Jennifer / Rubrum, Adam / Seiler, Patrick / Krauss, Scott / Webster, Robert / Webby, Richard J

    Vaccine

    2014  Volume 32, Issue 35, Page(s) 4571–4577

    Abstract: The H7N9 influenza virus caused significant mortality and morbidity in infected humans during an outbreak in China in 2013 stimulating vaccine development efforts. As previous H7-based vaccines have been poorly immunogenic in humans we sought to ... ...

    Abstract The H7N9 influenza virus caused significant mortality and morbidity in infected humans during an outbreak in China in 2013 stimulating vaccine development efforts. As previous H7-based vaccines have been poorly immunogenic in humans we sought to determine the immunogenic and protective properties of an inactivated whole virus vaccine derived from a 2013 H7N9 virus in ferrets. As whole virus vaccine preparations have been shown to be more immunogenic in humans, but less likely to be used, than split or surface antigen formulations, we vaccinated ferrets with a single dose of 15, 30, or 50 μg of the vaccine and subsequently challenged with wild-type A/Anhui/1/2013 (H7N9) either by direct instillation or by contact with infected animals. Although ferrets vaccinated with higher doses of vaccine had higher serum hemagglutinin inhibition (HI) titers, the titers were still low. During subsequent instillation challenge, however, ferrets vaccinated with 50 μg of vaccine showed no illness and shed significantly less virus than mock vaccinated controls. All vaccinated ferrets had lower virus loads in their lungs as compared to controls. In a separate study where unvaccinated-infected ferrets were placed in the same cage with vaccinated-uninfected ferrets, vaccination did not prevent infection in the contact ferrets, although they showed a trend of lower viral load. Overall, we conclude that inactivated whole-virus H7N9 vaccine was able to reduce the severity of infection and viral load, despite the lack of hemagglutinin-inhibiting antibodies.
    MeSH term(s) Animals ; Antibodies, Viral/blood ; Ferrets ; Hemagglutination Inhibition Tests ; Influenza A Virus, H7N9 Subtype/immunology ; Influenza Vaccines/administration & dosage ; Influenza Vaccines/immunology ; Lung/virology ; Orthomyxoviridae Infections/pathology ; Orthomyxoviridae Infections/prevention & control ; Severity of Illness Index ; Vaccines, Inactivated/administration & dosage ; Vaccines, Inactivated/immunology ; Viral Load ; Virus Shedding
    Chemical Substances Antibodies, Viral ; Influenza Vaccines ; Vaccines, Inactivated
    Language English
    Publishing date 2014-06-18
    Publishing country Netherlands
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 605674-x
    ISSN 1873-2518 ; 0264-410X
    ISSN (online) 1873-2518
    ISSN 0264-410X
    DOI 10.1016/j.vaccine.2014.06.016
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  6. Article ; Online: Chp1-Tas3 interaction is required to recruit RITS to fission yeast centromeres and for maintenance of centromeric heterochromatin.

    Debeauchamp, Jennifer L / Moses, Arian / Noffsinger, Victoria J P / Ulrich, Dagny L / Job, Godwin / Kosinski, Aaron M / Partridge, Janet F

    Molecular and cellular biology

    2008  Volume 28, Issue 7, Page(s) 2154–2166

    Abstract: The maintenance of centromeric heterochromatin in fission yeast relies on the RNA interference-dependent complexes RITS (RNA-induced transcriptional silencing complex) and RDRC (RNA-directed RNA polymerase complex), which cooperate in a positive feedback ...

    Abstract The maintenance of centromeric heterochromatin in fission yeast relies on the RNA interference-dependent complexes RITS (RNA-induced transcriptional silencing complex) and RDRC (RNA-directed RNA polymerase complex), which cooperate in a positive feedback loop to recruit high levels of histone H3 K9 methyltransferase activity to centromeres and to promote the assembly and maintenance of centromeric heterochromatin. However, it is unclear how these complexes are targeted to chromatin. RITS comprises Chp1, which binds K9-methylated histone H3; Ago1, which binds short interfering (siRNAs); the adaptor protein Tas3, which links Ago1 to Chp1; and centromeric siRNAs. We have generated mutants in RITS to determine the contribution of the two potential chromatin-targeting proteins Chp1 and Ago1 to the centromeric recruitment of RITS. Mutations in Tas3 that disrupt Ago1 binding are permissive for RITS recruitment and maintain centromeric heterochromatin, but the role of Tas3's interaction with Chp1 is unknown. Here, we define the Chp1 interaction domain of Tas3. A strain expressing a tas3 mutant that cannot bind Chp1 (Tas3(Delta)(10-24)) failed to maintain centromeric heterochromatin, with a loss of centromeric siRNAs, a failure to recruit RITS and RDRC to centromeres, and high levels of chromosome loss. These findings suggest a pivotal role for Chp1 and its association with Tas3 for the recruitment of RITS, RDRC, and histone H3 K9 methyltransferase activity to centromeres.
    MeSH term(s) Argonaute Proteins ; Carrier Proteins/genetics ; Carrier Proteins/physiology ; Cell Cycle Proteins/genetics ; Cell Cycle Proteins/physiology ; Centromere/ultrastructure ; Chromosomes, Fungal/ultrastructure ; Genes, Mating Type, Fungal/genetics ; Genomic Instability ; Heterochromatin/metabolism ; Heterochromatin/ultrastructure ; Histone Methyltransferases ; Histone-Lysine N-Methyltransferase/metabolism ; Multiprotein Complexes/metabolism ; Protein Interaction Mapping ; Protein Methyltransferases ; Protein Structure, Tertiary ; Protein Transport ; RNA, Fungal/metabolism ; RNA, Small Interfering/metabolism ; RNA-Binding Proteins ; Schizosaccharomyces/genetics ; Schizosaccharomyces/metabolism ; Schizosaccharomyces/ultrastructure ; Schizosaccharomyces pombe Proteins/genetics ; Schizosaccharomyces pombe Proteins/physiology ; Telomere/ultrastructure ; Transcription, Genetic
    Chemical Substances Ago1 protein, S pombe ; Argonaute Proteins ; Carrier Proteins ; Cell Cycle Proteins ; Chp1 protein, S pombe ; Heterochromatin ; Multiprotein Complexes ; RNA, Fungal ; RNA, Small Interfering ; RNA-Binding Proteins ; Schizosaccharomyces pombe Proteins ; Tas3 protein, S pombe ; Histone Methyltransferases (EC 2.1.1.-) ; Protein Methyltransferases (EC 2.1.1.-) ; Histone-Lysine N-Methyltransferase (EC 2.1.1.43)
    Language English
    Publishing date 2008-01-22
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 779397-2
    ISSN 1098-5549 ; 0270-7306
    ISSN (online) 1098-5549
    ISSN 0270-7306
    DOI 10.1128/MCB.01637-07
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