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[{"key": "dc.contributor.advisor", "value": "Vihinen-Ranta, Maija", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Paloheimo, Outi", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Niva, Juho", "language": "", "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2020-12-15T09:38:10Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2020-12-15T09:38:10Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2020", "language": "", "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/73210", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Enterovirus-sukuun kuuluva Coxsackievirus B3 aiheuttaa vakavia sairauksia mm. syd\u00e4nlihastulehdusta, kardiomyopatiaa ja tyypin 1 diabetesta. CVB3 on yksijuosteita RNA:ta sis\u00e4lt\u00e4v\u00e4 virus, joka replikoituu solun sytoplasmassa. CVB3 infektion loppuvaiheessa solun solukalvosta kuroutuu ulosp\u00e4in aktiinia sis\u00e4lt\u00e4vi\u00e4 pitk\u00e4nomaisia kalvoulkonemia. N\u00e4iden rakenteiden on arveltu mahdollistavan CVB3-virukselle ep\u00e4tyypillisen, ei-lyyttisen poistumisen is\u00e4nt\u00e4solusta ja levi\u00e4misen solusta toiseen. T\u00e4ss\u00e4 ty\u00f6ss\u00e4 tarkasteltiin viruksen solusta poistumista ja selvitettiin miten solutukirankaan vaikuttavat inhibiittorit s\u00e4\u00e4telev\u00e4t CVB3-infektion etenemist\u00e4 ja viruksen aiheuttamien kalvoulkonemien muodostumista green monkey kidney (GMK) soluissa. K\u00e4ytimme inhibiittoreita, jotka aiheuttavat aktiinitukirangan ja mikrotubulusten stabiloimisen tai depolymerisaation. Lis\u00e4ksi tutkimme miten myosiini II:n, myosin light chain -kinaasin (MLCK), RhoA-kinaasin (ROCK) ja MAP-kinaasin inhibitio vaikuttaa viruksen aiheuttamiin morfologisiin muutoksiin ja kalvoulkonemien muodostumiseen. Analysoimme CVB3-infektoituja ja inhibiittorik\u00e4siteltyj\u00e4 apinan munuaissoluja virtaussytometriaa ja konfokaalimikroskopiaa hy\u00f6dynt\u00e4en. Analyysiemme tulokset osoittivat, ett\u00e4 solutukirangan molekyyleja ja kinaaseja inhiboimalla, infektoituneiden GMK solujen m\u00e4\u00e4r\u00e4 pieneni merkitt\u00e4v\u00e4sti. Mikroskooppikuvista n\u00e4hd\u00e4\u00e4n, ett\u00e4 jotkut inhibiittorit (phalloidin ja nocodazole) my\u00f6s ehk\u00e4isiv\u00e4t CVB3-infektion aiheuttamia morfologisia muutoksia GMK soluissa. Lis\u00e4ksi huomasimme, ett\u00e4 ROCK:n ja MLCK:n inhibointi tai aktiinitukirangan stabiloiminen phalloidiinilla, mahdollisesti heikent\u00e4\u00e4 CVB3-partikkeleiden ei-lyyttist\u00e4 poistumista is\u00e4nt\u00e4solusta. Vaikka mikrotubulusten kemiallinen pirstominen v\u00e4hensi huomattavasti CVB3-infektoitujen solujen m\u00e4\u00e4r\u00e4\u00e4, ei mikrotubulusten inhibitio kuitenkaan est\u00e4nyt CVB3:n aiheuttamien kalvoulkonemien syntymist\u00e4 Tutkimustuloksemme tukee dynaamisen aktiinitukirangan merkityst\u00e4 CVB3:n ei-lyyttiselle poistumiselle.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Coxsackievirus B3 (CVB3) is an enterovirus that has an impact on severe diseases including myocarditis, cardiomyopathy, and type 1 diabetes. The infection of CVB3 has been noticed to alter the morphology of green monkey kidney cells by forming cage-like protrusions to the host cell membrane. The protrusions were found packed with actin filaments and viral capsids, thus, hypothesizing an atypical, non-lytic way of spreading from the host cell to another. To further examine the non-lytic pathway and the role of cellular cytoskeleton, stabilizing and depolymerizing inhibitors of actin and microtubules were used. The inhibition of myosin II, myosin light chain kinase (MLCK), RhoA-kinase (ROCK) and mitogen-activated kinase were also examined to see the structural and transcriptional effect on the morphological changes and protrusion formation. The CVB3-infected and drug-treated GMK cells were analyzed with fluorescent-activated cell sorting (FACS) and confocal laser scanning microscopy. Our analyses showed that inhibition of the cytoskeleton filaments and cell kinases decreases the number of CVB3 infected cells. According to the confocal microscopy images, the strongest inhibitors (phalloidin and nocodazole) also prevented some of the characteristic changes in the cell morphology normally seen in CVB3 infection. Our findings also suggest that inhibition of ROCK or MLCK or actin stabilizing with phalloidin could interfere with the nonlytic egress of CVB3. Although, microtubule depolymerization significantly inhibited the number of CVB3 infected cells, the microtubule inhibition did not prevent the emergence of CVB3-induced protrusions. Overall, our results were promising, and this work further establishes that a dynamic actin cytoskeleton plays an essential role in the possible non-lytic egress of CVB3.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by Paivi Vuorio (paelvuor@jyu.fi) on 2020-12-15T09:38:10Z\nNo. of bitstreams: 0", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2020-12-15T09:38:10Z (GMT). No. of bitstreams: 0\n Previous issue date: 2020", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "37", "language": "", "element": "format", "qualifier": "extent", "schema": "dc"}, {"key": "dc.format.mimetype", "value": "application/pdf", "language": null, "element": "format", "qualifier": "mimetype", "schema": "dc"}, {"key": "dc.language.iso", "value": "eng", "language": null, "element": "language", "qualifier": "iso", "schema": "dc"}, {"key": "dc.rights", "value": "In Copyright", "language": "en", "element": "rights", "qualifier": null, "schema": "dc"}, {"key": "dc.subject.other", "value": "coxsackievirus B3", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "cell cytoskeleton", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "egress", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "The role of cellular cytoskeleton in the egress of Coxsackievirus B3", "language": "", "element": "title", "qualifier": null, "schema": "dc"}, {"key": "dc.type", "value": "master thesis", "language": null, "element": "type", "qualifier": null, "schema": "dc"}, {"key": "dc.identifier.urn", "value": "URN:NBN:fi:jyu-202012157156", "language": "", "element": "identifier", "qualifier": "urn", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Pro gradu -tutkielma", "language": "fi", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.type.ontasot", "value": "Master\u2019s thesis", "language": "en", "element": "type", "qualifier": "ontasot", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Matemaattis-luonnontieteellinen tiedekunta", "language": "fi", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.faculty", "value": "Faculty of Sciences", "language": "en", "element": "contributor", "qualifier": "faculty", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Bio- ja ymp\u00e4rist\u00f6tieteiden laitos", "language": "fi", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.department", "value": "Department of Biological and Environmental Science", "language": "en", "element": "contributor", "qualifier": "department", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "Jyv\u00e4skyl\u00e4n yliopisto", "language": "fi", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "University of Jyv\u00e4skyl\u00e4", "language": "en", "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Solu- ja molekyylibiologia", "language": "fi", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "dc.subject.discipline", "value": "Cell and molecular biology", "language": "en", "element": "subject", "qualifier": "discipline", "schema": "dc"}, {"key": "yvv.contractresearch.funding", "value": "0", "language": "", "element": "contractresearch", "qualifier": "funding", "schema": "yvv"}, {"key": "dc.type.coar", "value": "http://purl.org/coar/resource_type/c_bdcc", "language": null, "element": "type", "qualifier": "coar", "schema": "dc"}, {"key": "dc.rights.accesslevel", "value": "openAccess", "language": null, "element": "rights", "qualifier": "accesslevel", "schema": "dc"}, {"key": "dc.type.publication", "value": "masterThesis", "language": null, "element": "type", "qualifier": "publication", "schema": "dc"}, {"key": "dc.subject.oppiainekoodi", "value": "4013", "language": "", "element": "subject", "qualifier": "oppiainekoodi", "schema": "dc"}, {"key": "dc.subject.yso", "value": "virukset", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "aktiinit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "viruses", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "actins", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.format.content", "value": "fulltext", "language": null, "element": "format", "qualifier": "content", "schema": "dc"}, {"key": "dc.rights.url", "value": "https://rightsstatements.org/page/InC/1.0/", "language": null, "element": "rights", "qualifier": "url", "schema": "dc"}, {"key": "dc.type.okm", "value": "G2", "language": null, "element": "type", "qualifier": "okm", "schema": "dc"}]
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