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[{"key": "dc.contributor.advisor", "value": "Sundberg, Lotta-Riina", "language": null, "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Goladze, Sophia", "language": null, "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "de Oliveira Patricio, Daniel", "language": null, "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Suokko, Valtteri", "language": null, "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2025-02-19T06:45:20Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2025-02-19T06:45:20Z", "language": null, "element": "date", "qualifier": "available", "schema": "dc"}, {"key": "dc.date.issued", "value": "2024", "language": null, "element": "date", "qualifier": "issued", "schema": "dc"}, {"key": "dc.identifier.uri", "value": "https://jyx.jyu.fi/handle/123456789/100262", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Antibiooteille vastustuskykyisten bakteerien yleistyminen on noussut yhdeksi suurimmista maailmanlaajuisista terveysongelmista. Se on aikaansaanut tarpeen katsoa perinteisten hoitomenetelmien ulkopuolelle ja kehitt\u00e4\u00e4 uusia menetelmi\u00e4 vastustuskykyisi\u00e4 bakteeri-infektioita vastaan. Faagiterapia eli bakteerien omien virusten k\u00e4ytt\u00e4minen is\u00e4nt\u00e4spesifisesti infektioita vastaan on yksi lupaavimmista menetelmist\u00e4. T\u00e4m\u00e4n tutkielman tavoitteena oli selvitt\u00e4\u00e4, miten limakalvoymp\u00e4rist\u00f6 vaikuttaa faagiresistenssin syntyyn Pseudomonas aeruginosa PA14 -bakteereissa ja miten syntynyt resistenssi vaikuttaa bakteerien virulenssiin. Lis\u00e4tavoitteena oli selvitt\u00e4\u00e4 musiinin vaikutusta PA14-bakteerien biofilmimuodostukseen. Resistenssikehitykselle optimaalisimmat ravinneolosuhteet ja aikajana m\u00e4\u00e4ritettiin kasvuanalyysill\u00e4 ja faagiresistentit PA14-bakteerit tuotettiin kasvattamalla bakteereita faagien DMS3vir ja GEC-PNG14 ollessa l\u00e4sn\u00e4. Resistenssimekanismeja vertailtiin spot-on-spot testill\u00e4 sek\u00e4 CRISPR PCR -kokeella. Lopuksi vastustuskykyisten bakteereiden virulenssi testattiin soluviljelymallilla hy\u00f6dynt\u00e4en ihmisen keuhkosolulinjan A549-soluja. Musiinin odotettiin edist\u00e4v\u00e4n DMS3vir-resistenssin synty\u00e4 CRISPR-j\u00e4rjestelm\u00e4n kautta, kun muutoin pintamodifikaatio olisi yleisemp\u00e4\u00e4. GEC-PNG14 oletettiin indusoivan resistenssi\u00e4 vain pintamodifikaation kautta sen anti-CRISPR-ominaisuuksien ansiosta. Resistenssin odotettiin laskevan bakteerien virulenssia riippumatta mekanismista. Musiini edisti faagiresistenssikehityst\u00e4 ja alensi bakteerien biofilmimuodostuskyky\u00e4, mutta ei vaikuttanut resistenssi-mekanismeihin. Resistenttien bakteerien virulenssi oli kuitenkin merkitt\u00e4v\u00e4sti matalampi kuin villityypin bakteereilla. CRISPR-puolustuksen puutoksen syyt eiv\u00e4t selvinneet t\u00e4ss\u00e4 tutkielmassa, mutta j\u00e4rjestelm\u00e4 saattoi vain esimerkiksi olla ep\u00e4suotuisa pintamodifikaatioon verrattuna faagi-is\u00e4nt\u00e4- ja ymp\u00e4rist\u00f6tekij\u00f6iden takia. Virulenssimuutokset johtuivat luultavimmin pintamodifikaatioista, kun bakteerit muokkasivat solukalvon reseptoreitaan saavuttaakseen resistenttiyden. Viimeisimp\u00e4n\u00e4, biofilmimuodostuksen muutokset johtuivat luultavimmin musiinin kyvyst\u00e4 edist\u00e4\u00e4 bakteerien liikehdint\u00e4\u00e4.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "The rise of antibiotic resistance in bacteria is one of the severest global health hazards, forcing us to investigate novel treatment methods against resistant pathogens. Phage therapy, the use of bacterial viruses to selectively kill their hosts, is among the most feasible of the alternatives. This thesis aimed to determine how a mucous environment, simulated with mucin, effects phage resistance development in Pseudomonas aeruginosa PA14, and how said resistance affects bacterial virulence. Additionally, how mucin affects PA14 biofilm formation was assessed. A growth rate analysis was first used to assess the best nutrient conditions and timescale for phage resistance generation. Next, phage resistant PA14 were produced in an evolution experiment using two phages: DMS3vir and GEC-PNG14. Resistance mechanisms were evaluated using a spot-on-spot assay and CRISPR PCR. Lastly, virulence changes caused by resistance generation were quantified by cell viability assays utilizing human A549 cells. Mucin was expected to promote resistance to DMS3vir through the CRISPR system while surface modification would reign in its absence. GEC-PGN14 was expected to induce resistance only through surface modification due to its anti-CRISPR capabilities. The virulence of the resistant mutants was expected to be lowered compared to wild type bacteria regardless of the type of resistance. Mucin was also expected to increase biofilm production in PA14. Ultimately, mucin promoted phage resistance generation and inhibited biofilm formation, but did not influence the phage resistance mechanisms. Phage resistant mutants had a significantly lowered virulence compared to wild type bacteria. The reason for the lack of CRISPR defences was not identified, although the system might just have been costly compared to surface modification due to phage-host and environmental characteristics. The observed virulence changes may have stemmed from changes in motility and interaction capabilities, with surface mutants sacrificing cell surface receptors in exchange for resistance. Lastly, changes in biofilm formation resulted probably from mucin promoting twitching motility in PA14.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by Paivi Vuorio (paelvuor@jyu.fi) on 2025-02-19T06:45:20Z\nNo. of bitstreams: 0", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2025-02-19T06:45:20Z (GMT). No. of bitstreams: 0\n Previous issue date: 2024", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "44", "language": null, "element": "format", "qualifier": "extent", "schema": "dc"}, {"key": "dc.language.iso", "value": "eng", "language": null, "element": "language", "qualifier": "iso", "schema": "dc"}, {"key": "dc.rights", "value": "In Copyright", "language": null, "element": "rights", "qualifier": null, "schema": "dc"}, {"key": "dc.subject.other", "value": "CRISPR-Cas", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "ESKAPE", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "mucin", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "surface modification", "language": null, "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "Effect of mucosal environment on phage resistance and subsequent infectivity of Pseudomonas aeruginosa", "language": null, "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-202502192040", "language": null, "element": "identifier", "qualifier": "urn", "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": null, "element": "contributor", "qualifier": "organization", "schema": "dc"}, {"key": "dc.contributor.organization", "value": "University of Jyv\u00e4skyl\u00e4", "language": null, "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": "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.yso", "value": "fagiterapia", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "antibioottiresistenssi", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.rights.url", "value": "https://rightsstatements.org/page/InC/1.0/", "language": null, "element": "rights", "qualifier": "url", "schema": "dc"}]
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