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[{"key": "dc.contributor.advisor", "value": "Kalkkinen, Nisse", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Valmu, Leena", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.advisor", "value": "Yl\u00e4nne, Jari", "language": "", "element": "contributor", "qualifier": "advisor", "schema": "dc"}, {"key": "dc.contributor.author", "value": "Karinen, Anu", "language": "", "element": "contributor", "qualifier": "author", "schema": "dc"}, {"key": "dc.date.accessioned", "value": "2021-01-04T06:43:37Z", "language": null, "element": "date", "qualifier": "accessioned", "schema": "dc"}, {"key": "dc.date.available", "value": "2021-01-04T06:43:37Z", "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/123456789/73520", "language": null, "element": "identifier", "qualifier": "uri", "schema": "dc"}, {"key": "dc.description.abstract", "value": "V\u00e4lit\u00f6n tyypin I luonnonkumiallergia on IgE-v\u00e4litteinen allerginen reaktio, jota esiintyy s\u00e4\u00e4nn\u00f6llisesti luonnonkumituotteiden kanssa kosketuksiin joutuvilla henkil\u00f6ill\u00e4. Luonnonkumiallergia aiheutti varsinkin 1990-luvulla vakavia ongelmia erityisesti terveydenhuollon ty\u00f6ntekij\u00f6ille ja potilaille, jotka ovat syntym\u00e4st\u00e4\u00e4n l\u00e4htien l\u00e4pik\u00e4yneet lukuisia kirurgisia toimenpiteit\u00e4, kuten selk\u00e4rankahalkiolapset. Vaikka huomattavaa edistymist\u00e4 on tapahtunut luonnonkumiallergian tunnistamisessa ja ehk\u00e4isyss\u00e4, voi se yh\u00e4 olla kehittyviss\u00e4, mutta my\u00f6s teollistuneissa maissa merkitt\u00e4v\u00e4 ty\u00f6per\u00e4inen sairaus tai aiheuttaa allergisia reaktioita riskiryhmiin kuuluvilla, jotka k\u00e4ytt\u00e4v\u00e4t luonnonkumisia l\u00e4\u00e4kinn\u00e4llisi\u00e4 tai kotitaloustuotteita. T\u00e4h\u00e4n menness\u00e4 15 luonnonkumin proteiinia on saanut allergeeniluokituksen. N\u00e4ist\u00e4 yksi, Hevb2 (\uf062-1,3-glukanaasi), on tunnistettu ja kloonattu, mutta sen merkitys allergeenina on j\u00e4\u00e4nyt ep\u00e4selv\u00e4ksi. T\u00e4ss\u00e4 tutkimuksessa Hevb2-proteiini puhdistettiin ultrasentrifugoidun luonnonkumin pohjafraktiosta (B-seerumi) kationinvaihto-, geelisuodatus- ja k\u00e4\u00e4nteisfaasikromatografialla. Proteiinin tunnistamisessa ja sen rakenteen karakterisoimisessa k\u00e4ytettiin geelielektroforeesia, MALDI-TOF MS (matriisiavusteinen laserdesorptio-ionisaatio-lentoaika massaspektrometria), ESI (s\u00e4hk\u00f6sumutusionisaatio) MS ja MS/MS -menetelmi\u00e4 sek\u00e4 N-terminaalista sekvensointia. Immunologinen karakterisointi suoritettiin ELISA (entsyymiv\u00e4litteinen immunosorbenttim\u00e4\u00e4ritys) ja Western blot -menetelmill\u00e4 k\u00e4ytt\u00e4m\u00e4ll\u00e4 suomalaisten potilaiden seerumeja, joista muutama oli per\u00e4isin my\u00f6s luonnonkumiallergisilta potilailta. Hevb2-proteiinin havaittiin esiintyv\u00e4n kahtena, molekyylipainoltaan 36,4 ja 37,6 kDa kokoisena isomuotona. Isomuodot tunnistettiin ja niiden yhteys m\u00e4\u00e4ritettiin massaspektrometrisesti peptidisormenj\u00e4lkien avulla. Molemmat olivat N-terminaalisesti blokattuja, jonka aiheutti 36,4 kDa isomuodolla syklinen pyroglutamiinihappo. ELISA-immunom\u00e4\u00e4rityksess\u00e4 vain 10 %:lla (7/71) potilaista l\u00f6ytyi spesifist\u00e4 IgE-vasta-ainetta 36,4 kDa Hevb2-proteiinille. Vaikka t\u00e4m\u00e4n alustavan kokeen perusteella Hevb2-proteiini ei vaikuttaisi olevan merkitt\u00e4v\u00e4 luonnonkumiallergeeni suomalaisilla potilailla, tutkimus on syyt\u00e4 toistaa valikoituneemmalla potilas- ja kontrollijoukolla niin, ett\u00e4 molemmat isomuodot analysoidaan yhdess\u00e4 ja proteiinin\u00e4ytteen puhtaus varmistetaan viel\u00e4 esimerkiksi saatavilla olevilla monoklonaalisilla vasta-aineilla.", "language": "fi", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.abstract", "value": "Type \uf049 immediate hypersensitivity to natural rubber latex (NRL) is an IgE-mediated allergic reaction recognized in individuals who are frequently in contact with NRL products. NRL allergy has particularly in the 1990s caused the serious problems especially among health care workers and patients who have undergone multiple surgical operations, such as spina bifida children. Although substantial progress has been made in the understanding and prevention of NRL allergy, it may still be an important disease in developing but also in industrialized countries among person with occupational exposure to NRL gloves and among individuals belonging to risk group and using latex-containing medical or household products. To date, 15 latex proteins have received an international nomenclature designation as allergens. One of these, Hevb2 (\uf062-1,3-glucanase) has been identified and cloned but its significance as latex allergen has remained ambiguous. In this study, Hevb2 was purified from the bottom fraction (B-serum) of ultracentrifuged NRL by cation-exchange chromatography, gel filtration and reversed-phase chromatography. Identification and characterization of Hevb2 was performed using gel electrophoresis, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS), electrospray ionization (ESI) MS and MS/MS as well as N-terminal sequencing. Immunological characterization of Hevb2 was performed by enzyme-linked immunosorbent assay (ELISA) and immunoblotting (Western blotting) using sera of Finnish patients, including also a few NRL allergic patients. Hevb2 protein occurred in two isoforms, 36.4 and 37.6 kDa according to mass spectrometry. The identity and the relationship of these two isoforms was demonstrated by peptide mass fingerprinting. Both isoforms were found to be N-terminally blocked but only the blocking group of 36.4 kDa Hevb2 was demonstrated to be a cyclic pyroglutamic acid. In ELISA, only 10 % (7/71) of sera from Finnish patients showed specific IgE antibodies to purified 36.4 kDa Hevb2. While this preliminary study demonstrated that highly purified 36.4 kDa Hevb2 appears not to be major latex allergen among Finnish population, more study is still needed with more selected patient and control group, and performing experiment so that the both isoforms are analysed together and the purity of protein preparation is further ascertained e.g. by monoclonal antibodies.", "language": "en", "element": "description", "qualifier": "abstract", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Submitted by Paivi Vuorio (paelvuor@jyu.fi) on 2021-01-04T06:43:37Z\nNo. of bitstreams: 0", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.description.provenance", "value": "Made available in DSpace on 2021-01-04T06:43:37Z (GMT). No. of bitstreams: 0\n Previous issue date: 2020", "language": "en", "element": "description", "qualifier": "provenance", "schema": "dc"}, {"key": "dc.format.extent", "value": "117", "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": "fin", "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": "beeta-1", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "3-glukanaasi", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "B-seerumi", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "Hevea brasiliensis", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "IgE-vasta-aine", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.subject.other", "value": "luonnonkumiallergia", "language": "", "element": "subject", "qualifier": "other", "schema": "dc"}, {"key": "dc.title", "value": "Kumipuun Hevb2-proteiinin puhdistus, tunnistus ja karakterisointi sek\u00e4 merkitys luonnonkumiallergeenina", "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-202101041001", "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.collaborator", "value": "public", "language": "", "element": "contractresearch", "qualifier": "collaborator", "schema": "yvv"}, {"key": "yvv.contractresearch.funding", "value": "0", "language": "", "element": "contractresearch", "qualifier": "funding", "schema": "yvv"}, {"key": "yvv.contractresearch.initiative", "value": "student", "language": "", "element": "contractresearch", "qualifier": "initiative", "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": "nestekromatografia", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "massaspektrometria", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "allergeenit", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "allergia", "language": null, "element": "subject", "qualifier": "yso", "schema": "dc"}, {"key": "dc.subject.yso", "value": "lateksit", "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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