Kvantifikacija fekalnog eozinofilnog kationskog proteina

master's thesis

• Title (english): Quantification of eosinophil cationic protein in feces
• Author: Ivana Ćaleta
• Mentor: Karmela Barišić (mentor)
• Committee member: Karmela Barišić (predsjednik povjerenstva)
• Committee member: Roberta Petlevski (član povjerenstva)
• Committee member: Renata Zrinski Topić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2018-09-28, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

ECP je bazični protein smješten u granulama eozinofilnih granulocita koji je uključen u sustav imunosnog odgovora. U stanicama domaćina broj eozinofila korelira s koncentracijama eozinofilnog kationskog proteina (ECP) koji se oslobađa njihovom aktivacijom. Karakteristika ECP-a jest citotoksičnost i sudjelovanje u različitim upalnim procesima. Intestinalna sluznica pacijenata s upalnom bolesti crijeva (IBD) karakterizirana je oštećenjem epitelnih stanica i infiltracijom različitih upalnih stanica, uključujući i ECP. Cilj ovoga diplomskoga rada bio je ispitati učinkovitost i predložiti potencijalne modifikacije metode za kvantifikaciju fekalnog ECP-a. Za bolju ekstrakciju fekalnog ECP-a korišten je ekstrakcijski pufer s CTAB-om. Ispitane su modifikacije originalnog postupka ekstrakcije ECP-a u pogledu priređivanja homogenata uzorka fecesa i trajanja inkubacije. Također je ispitana stabilnost i ponovljivost mjerenja ECP-a. Uporabom instrumenta Phadia100 provedeno je ImmunoCAP ECP mjerenje koncentracija ECP-a u uzorcima. Bolji rezultati postignuti su homogenizacijom uzoraka ultrazvučnim sonifikatorom, te sekundarnim razrjeđenjem od 5 i 10 puta. Stabilnost uzoraka osigurana je pohranom u zamrzivaču na -20°C prije analiza.

Abstract (english)

ECP is a basic protein localized in granules of the eosinophil granulocytes and is included in immune response reactions. In host cells, eosinophil levels correlate with ECP levels that are released during their activation. Characteristic of ECP is cytotoxicity and involvement in various inflammatory processes. Intestinal mucose of the patients with IBD is characterized with damaged epitelial cells and infiltration of various inflammatory cells, including eosinophils. The goal of this research was to examine efficacy and suggest potential modifications of the method for the quantification of the ECP in stool samples. For better fecal ECP extraction, buffer mixture with CTAB was used. Modifications of the original ECP extraction procedure were examined in terms of stool samples preparation and incubation time. Stability and repeatability of ECP measurement was also tested. ImmunoCAP ECP assay preformed with Phadia100 instrument was used for measuring ECP concentrations in samples. Better results were achieved by sample homogenization with ultrasonic sonificator and secundary dilution of 5 x and 10 x. Sample stability was secured by storing in the freezer at the -20°C before assays.

• Keywords: eozinofili; ECP; IBD; metoda kvantifikacije u fecesu; imunoanaliza
• Keywords (english): eosinophil; ECP; IBD; quantification method in feces; immunoassays
• Language: croatian
• URN:NBN: urn:nbn:hr:163:596480
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2019-09-26 13:21:04