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[{"key": "dc.contributor.advisor", "value": "Turkki, Paula", "language": null, "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Marjom\u00e4ki, Varpu", "language": null, "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Sallinen-Dal Maso, Heidi", "language": null, "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2017-06-08T16:43:34Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2017-06-08T16:43:34Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2017", "language": null, "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.other", "value": "oai:jykdok.linneanet.fi:1703983", "language": null, "element": "identifier", "qualifier": "other", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/54383", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Enterovirukset ovat pieni\u00e4, vaipattomia, yksijuosteisia RNA-viruksia, jotka infektoivat yleisesti ihmisi\u00e4. Ne aiheuttavat laajan kirjon lievi\u00e4 sek\u00e4 vakavia oireita ett\u00e4 taudinkuvia, muun muassa flunssan oireita, syd\u00e4nlihastulehdusta sek\u00e4 kroonisia tauteja. Tyypillisesti enterovirusinfektio johtaa nopeaan solun hajoamiseen ja viruspartikkelien vapautumiseen solukuoleman kautta. Tutkimustieto viittaa kuitenkin siihen, ett\u00e4 enterovirukset kykenisiv\u00e4t my\u00f6s pysyv\u00e4\u00e4n infektiomuotoon, jossa soluviljelm\u00e4 pysyy elossa virusinfektiosta huolimatta. T\u00e4t\u00e4 infektiomuotoa ei kuitenkaan ole viel\u00e4 paljoa tutkittu eik\u00e4 esimerkiksi tiedet\u00e4 miten infektio n\u00e4ist\u00e4 soluista levi\u00e4\u00e4. Pysyv\u00e4 enterovirusinfektio on yhdistetty esimerkiksi ykk\u00f6styypin diabetekseen tehden siit\u00e4 ajankohtaisen tutkimuskohteen. Ty\u00f6n tarkoituksena oli tutkia kahta eri coxsackievirusperheeseen kuuluvaa enterovirusta (CVB3, CVB5) kahdessa erilaista solutyyppi\u00e4 edustavassa ihmissolulinjassa (A549, RD). Virusinfektioista haluttiin m\u00e4\u00e4ritt\u00e4\u00e4 uusien viruspartikkeleiden muodostumisen sek\u00e4 genomin monistumisen kinetiikkaa, solukuoleman ajoitusta ja infektion levi\u00e4mismekanismia. Luminesenssiin perustuvat el\u00e4vyysmittaukset osoittivat, ett\u00e4 molemmat virukset tappoivat A549-soluviljelm\u00e4t hyvinkin nopeasti CVB3 24 tuntiin menness\u00e4 ja CVB5 72 tuntiin menness\u00e4 osoittaen kyseess\u00e4 olevan solut tappava infektio. RD-soluviljelm\u00e4t pysyiv\u00e4t elossa useita p\u00e4ivi\u00e4 virusinfektiosta huolimatta viitaten kyseess\u00e4 olevan pysyv\u00e4 infektiomalli. Immunofluoresenssileimaukset ja konfokaalimikroskopia osoittivat, ett\u00e4 A549-soluissa molemmilla viruksilla genomien monistuminen sek\u00e4 uusien viruspartikkelien tuotto oli tehokasta. Ajallisesti virukset kuitenkin k\u00e4ytt\u00e4ytyiv\u00e4t eri tavalla: CVB5 oli huomattavasti hitaampi merkitt\u00e4v\u00e4n virustuoton ilmetess\u00e4 vasta 12 tuntia infektion aloituksesta, kun taas CVB3:lla sama tilanne n\u00e4htiin jo kuusi tuntia infektion aloituksesta. RD-soluissa molemmat virukset olivat hitaampia kuin A549-soluissa. Lis\u00e4ksi vain pieni osa soluista vaikutti infektoituvan viruksilla mik\u00e4 n\u00e4kyi RD-soluissa CVB3:lla alle 20 % ja CVB5:ll\u00e4 alle 10 % infektiotehokkuutena 6-96 tunnin aikav\u00e4lill\u00e4 kun tarkasteltiin virusgenomin monistumista. Kapsidien tuotto seurasi t\u00e4t\u00e4 samaa linjaa CVB3:lla kun taas CVB5:ll\u00e4 alle 20 % soluista n\u00e4ytti olevan positiivisia virustuoton suhteen aikav\u00e4lill\u00e4 6-48 tuntia kunnes yll\u00e4tt\u00e4en 96 tuntia infektiosta CVB5-kapsidille positiivisia RD-soluja olikin 58 % populaatiosta. T\u00e4m\u00e4 yll\u00e4tt\u00e4v\u00e4 piikki ei ollut n\u00e4kyviss\u00e4 virusgenomin monistumispositiivisissa soluissa tarkoittaen, ett\u00e4 n\u00e4m\u00e4 olivat luultavimmin virusgenomin monistumista ilment\u00e4vist\u00e4 soluista siirtyneit\u00e4 viruksia. Samaa ilmi\u00f6t\u00e4 ei havaittu CVB3-infektiossa viitaten eroavaisuuksiin virusten levi\u00e4mismekanismeissa t\u00e4ss\u00e4 solulinjassa. My\u00f6s kapsidileiman ja tuotettujen kapsidien sijainti soluissa oli erilainen CVB3- ja CVB5-infektioissa, mik\u00e4 viittaa siihen, ett\u00e4 n\u00e4ill\u00e4 viruksilla on eri levi\u00e4misstrategiat. Lis\u00e4ksi alustavat kokeet antoivat viitteit\u00e4, ett\u00e4 RD-solut eritt\u00e4v\u00e4t viruksia CVB3- ja CVB5-infektioiden aikana ilman solukuolemaa. Kaiken kaikkiaan ty\u00f6ss\u00e4 havaittiin kahta erilaista infektiomekanismia: tappavaa ja pysyv\u00e4\u00e4. Molemmissa malleissa havaittiin viruksen siirtymist\u00e4 uusiin soluihin ilman solujen hajoamista mahdollisesti kalvorakkulav\u00e4litteisesti. RD-solujen pysyv\u00e4 CVB5-infektio kaipaisi lis\u00e4kokeita, jotta ymm\u00e4rrett\u00e4isiin uusien viruspartikkeleiden siirtyminen valtaosaan soluista hyvinkin yht\u00e4kkisesti useita p\u00e4ivi\u00e4 infektion aloituksen j\u00e4lkeen.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Enteroviruses are small, non-enveloped, single-stranded RNA-viruses. They are one of the most common human pathogens. They cause a wide variety of mild and severe symptoms and diseases, such as common cold, myocarditis and different chronic diseases. Typically enterovirus infection quickly leads to the release of virus particles via cell death called cell lysis. However, research indicates that enteroviruses are also capable of causing persistent infection in which the cell culture stays alive despite of virus infection. This form of infection is not yet widely studied and for example it is not known how the infection spreads from these cells. Persistent enterovirus infection has been linked to type I diabetes making it an interesting research subject. The goal of this study was to examine two different enteroviruses (CVB3, CVB5) that belong to the family of coxsackieviruses, in two human cell lines that represent two different cell types (A549, RD). The goal was to determine the kinetics of forming new virus particles and replication, the timing of the cell death and the spreading mechanism of the infection. Cell viability test based on luminescence showed that both viruses killed A549 cell cultures quickly: CVB3 by 24 hours and CVB5 by 72 hours. This showed that the infection in question was lytic. RD cell cultures stayed alive for several days despite of the virus infection which indicates persistent infection model. Immunofluorescence labelling and confocal microscopy showed that in A549 cells the replication and the production of new virus particles were efficient with both viruses despite the viruses behaving differently in regards of time. CVB5 was considerably slower with significant virus production appearing only 12 hours post infection. The CVB3 was in a similar situation already in 6 hours post infection. In RD cells both viruses were slower than in A549 cells. Only a subpopulation off RD cells seemed to be infected with the viruses. This was shown in CVB3 with less than 20 % and in CVB5 with less than 10 % infected cells in a time span of 6-96 hours during which the replication was examined. Similar results were shown with production of capsids in CVB3. In CVB5 infection less than 20 % of cells showed positive for capsids labeling during a time span of 6-48 hours. Surprisingly though, 96 hours post infection 58 % of the RD cell population were CVB5 capsid positive cells. This surprising spike was not seen in the replication positive cells, which means that these were probably viruses transferred from the cells that show replication. This phenomenon was not seen during CVB3 infection meaning that the viruses have different spreading mechanisms in this cell line. The location of the capsids label was also different in CVB3 and CVB5 infections suggesting that these viruses have different spreading mechanisms. Additionally, preliminary tests indicate that RD cells produce viruses during CVB3 and CVB5 infections without cell lysis. All in all, in this study two different infection mechanisms (lytic and persistent) were examined. In both models viruses were observed to transfer to new cells without cell death, possibly via vesicle spreading. The persistent CVB5 infection of RD cells needs additional testing as to understand the spreading of new virus particles into majority of cells suddenly several days post infection.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted using Plone Publishing form by Heidi Sallinen-Dal Maso (hepasall) on 2017-06-08 16:43:33.802878. Form: Pro gradu -lomake (https://kirjasto.jyu.fi/julkaisut/julkaisulomakkeet/pro-gradu-lomake). JyX data: [jyx_publishing-allowed (fi) =False]", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by jyx lomake-julkaisija (jyx-julkaisija.group@korppi.jyu.fi) on 2017-06-08T16:43:34Z\nNo. of bitstreams: 2\nURN:NBN:fi:jyu-201706082754.pdf: 2249987 bytes, checksum: a08a506475cabec75774d5446af81f9a (MD5)\nlicense.html: 1147 bytes, checksum: d64fa6ead9cb8d2cbcaa5d6bc1b91cd8 (MD5)", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2017-06-08T16:43:34Z (GMT). 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Aineisto on luettavissa Jyv\u00e4skyl\u00e4n yliopiston kirjaston arkistoty\u00f6asemalta. Ks. https://kirjasto.jyu.fi/fi/tyoskentelytilat/laitteet-ja-tilat.", "language": "fi", "element": "rights", "qualifier": "accessrights", "schema": "dc"}, {"key": "dc.rights.accessrights", "value": "This material has a restricted access due to copyright reasons. It can be read at the workstation at Jyv\u00e4skyl\u00e4 University Library reserved for the use of archival materials: https://kirjasto.jyu.fi/en/workspaces/facilities.", "language": "en", "element": "rights", "qualifier": "accessrights", "schema": "dc"}, {"key": "dc.type.okm", "value": "G2", "language": null, "element": "type", "qualifier": "okm", "schema": "dc"}]
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