Klinička primjena mjerenja hitotriozidaze u plazmi i suhoj kapi krvi na filtar papiru

master's thesis

• Title (english): Clinical significance of chitotriosidase in plasma and dried blood spot
• Author: Ines Fijan
• Mentor: Ksenija Fumić (mentor)
• Committee member: Ksenija Fumić (predsjednik povjerenstva)
• Committee member: Sandra Šupraha Goreta (član povjerenstva)
• Committee member: Marija Grdić Rajković (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2017-01-27, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Nasljedni metabolički poremećaji su monogenske bolesti, kod kojih se kao posljedica mutacije
gena javlja sinteza nefunkcionalnog enzima ili njegova nedostatna sinteza. Zbog toga može doći
do nakupljanja supstrata ili jednog od međuprodukata metaboličkog puta. Lizosomske bolesti
nakupljanja spadaju u skupinu nasljednih metaboličkih poremećaja. Gaucherova bolest je najčešća
lizosomska bolest. Riječ je o sfingolipidozi uzrokovanoj deficijencijom lizosomalnog enzima β-
glukocerebrozidaza. Kao rezultat nedostatka β-glukocerebrozidaze dolazi do akumulacije
glukozilceramida u lizosomima makrofaga te se tako formiraju Gaucherove stanice u kojima se
nakuplja enzim hitotriozidaza. Stoga je hitotriozidaza biomarker koji se može određivati kod
Gaucherovih bolesnika. Također smatra se korisnim biomarkerom i u sarkoidozi, multiploj
sklerozi te drugim patološkim stanjima. U ovom diplomskom radu mjerila se aktivnost enzim
hitotriozidaze u 84 uzorka plazme bolničkih pacijenata te suhe kapi krvi na filtar papiru.
Usporedbom rezultata dobivenih u obje vrste uzoraka cilj je bio razviti metodu za određivanje
hitotriozidaze u suhoj kapi krvi na filtar papiru jer ona kao uzorak ima veliki broj prednosti u
odnosu na plazmu. Nakon uspostavljanja zadovoljavajućih uvjeta mjerenja hitotriozidaze u suhoj
kapi krvi na filtar papiru i njezinim mjerenjem spektrofluorimetrijom, dobiveni otkloni
fluorescencije pokazali su da korelacija između dobivenih rezultata u suhoj kapi krvi na filter
papiru i rezultata u uzorcima plazme s povišenom aktivnosti ne postoji.

Abstract (english)

Hereditary disorders are monogenetic diseases. In this type of disease synthesis of disfunctional
enzyme or lack of synthesised enzyme are caused by mutation of a gene. Moreover, those diseases
can cause accumulation of certain substrates or intermediates of metabolic pathways. Lysosomal
storage disorder is one of the types of this hereditary disorders. Most common one is called
Gauche disease. It is a sphingolipidosis caused by the deficiency of glucocerebrosidase.
Consequent accumulation of glucosylceramide inside the lysosomes of macrophages results in
formation of so called Gaucher cells. Chitotriosidase (CT) is enzyme that can be determined as
biomarker in patients with Gaucher disease because it accumulates in macrophages. Moreover, CT
is also a useful biomarker in sarcoidosis, multiple sclerosis and samo other conditions. In this work
we carried out the analysis of 84 samples of plasma and dried blood spot. In both sample types we
measured CT activity.. The goal was to develop a method for determination of CT in dried blood
spot, which has many advantages compared to plasma samples.
Therefore, after we achieved appropriate conditions for CT determination in dried blood spot we
measured CT activity in both samples for each patient using flourescence spectroscopy.
Fluorescence readings showed no correlation among dried blood spot samples and plasma samples
in which we expected high CT activity.

• Keywords: suha kap krvi na filter papiru; Gaucherova bolest; hitotriozidaza
• Keywords (english): dried blood spot; Gaucher disease; chitotriosidase
• Language: croatian
• URN:NBN: urn:nbn:hr:163:184849
• Study programme: Title: Medical biochemistry; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra medicinske biokemije (magistar/magistra medicinske biokemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-09-14 10:23:28