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Razvoj metode izolacije inhibitorjev trombina iz glive stereum hirsutum = Development of isolation method for inhibitors thrombin from fungus stereum hirsutum : diplomska nalogaBorštar Dim, KatarinaBolezni srca in ozilja so zelo pogosta obolenja in predstavljajo enega najpogostejsih vzrokov smrti sodobnega èloveka. Trombin ima kot serinska proteaza pri tem kljuèno vlogo, saj sodeluje pri ... procesih strjevanja krvi in razvoju tromboze. Zaradi tega predstavlja primerno tarèo za inhibicijo. Glive so farmakolosko gledano relativno neraziskano podroèje, zato je iskanje aktivnih substanc v njih obetavno. Resetanje nekaterih izbranih vrst gliv glede inhibitorne aktivnosti na trombin je pokazalo, da dlakava slojevka (Stereum hirsutum) zmanjsuje aktivnost trombina. Na osnovi teh rezultatov smo se odloèili razviti metodo izolacije aktivne komponente iz glivnega materiala.Dlakava slojevka je v zadnjem obdobju tudi sicer delezna poveèanega znanstvenega zanimanja. Metanolni izvleèek dlakave slojevke smo pripravili iz 1 kg droge in ga macerirali v 6 L metanola. Po odparitvi topila smo suh izvleèek raztopili v ustrezni mobilni fazi in ga uporabili za kolonsko in tankoplastno kromatografijo. Po razvijanju vzorca v sedmih mobilnih fazah, v razliènih razmerjih diklormetana in metanola, smo se odloèili za mobilno fazo diklormetanČmetanol = 5 Č 1. Inhibitorno aktivnost trombina smo spremljali s pomoèjo spektrofotometra na mikrotitrskih plosèicah, na katerih smo merili absorbanco produkta, ki nastaja po amidolitièni cepitvi substrata S-2238. Med razvojem metode za izolacijo smo spremljali inhibitorno aktivnost posamezne frakcije in nadaljevali z najbolj aktivno frakcijo. Rezultati meritve inhibicije na trombin kazejo ponovljive inhibitorne aktivnosti. Pri zadnjih dveh kolonskih kromatografijah smo uporabili mobilno fazo diklormetanČmetanol = 20 Č 1, da bi dobili bolj selektivno loèene lise, a smo ugotovili, da imamo se vedno prisotnih veliko lis, ki v prejsnji mobilni fazi niso bile vidne. Dobro bi bilo poiskusiti se druge kromatografske tehnike, ki bi dale veè rezultatov o fizikalno-kemijskih lastnostih aktivne spojine.Type of material - undergr. thesisPublication and manufacture - Ljubljana : [Borštar D. K.], 2002Language - slovenianCOBISS.SI-ID - 1194097
Author
Borštar Dim, Katarina
Other authors
Štrukelj, Borut |
Doljak, Bojan
Topics
hemostaza |
proteaze |
kromatografija |
glive |
metode |
peptidaze |
priprava vzorcev
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Library | Call number – location, accession no. ... | Copy status |
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Faculty of Pharmacy, Lj. | Knjižnica dp 581 Borštar Dim K. Razvoj IN: 0006584 |
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Štrukelj, Borut | 07849 |
Doljak, Bojan | 19785 |
Source: Personal bibliographies
and: SICRIS
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