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master's thesis
Fluorimetrijska metoda za mjerenje aktivnosti tripeptidil peptidaze 1 u homogenatu leukocita
Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry, 2019. urn:nbn:hr:163:125241
Blažević, Nikolina
University of Zagreb
Faculty of Pharmacy and Biochemistry
Department of medical biochemistry and haematology

Institutional repository: Repository of Faculty of Pharmacy and Biochemistry University of Zagreb

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Blažević, N. (2019). Fluorimetrijska metoda za mjerenje aktivnosti tripeptidil peptidaze 1 u homogenatu leukocita (Master's thesis). Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry. Retrieved from https://urn.nsk.hr/urn:nbn:hr:163:125241

Blažević, Nikolina. "Fluorimetrijska metoda za mjerenje aktivnosti tripeptidil peptidaze 1 u homogenatu leukocita." Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, 2019. https://urn.nsk.hr/urn:nbn:hr:163:125241

Blažević, Nikolina. "Fluorimetrijska metoda za mjerenje aktivnosti tripeptidil peptidaze 1 u homogenatu leukocita." Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, 2019. https://urn.nsk.hr/urn:nbn:hr:163:125241

Blažević, N. (2019). 'Fluorimetrijska metoda za mjerenje aktivnosti tripeptidil peptidaze 1 u homogenatu leukocita', Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, accessed 06 May 2024, https://urn.nsk.hr/urn:nbn:hr:163:125241

Blažević N. Fluorimetrijska metoda za mjerenje aktivnosti tripeptidil peptidaze 1 u homogenatu leukocita [Master's thesis]. Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry; 2019 [cited 2024 May 06] Available at: https://urn.nsk.hr/urn:nbn:hr:163:125241

N. Blažević, "Fluorimetrijska metoda za mjerenje aktivnosti tripeptidil peptidaze 1 u homogenatu leukocita", Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, 2019. Available at: https://urn.nsk.hr/urn:nbn:hr:163:125241

Metadata
TitleFluorimetrijska metoda za mjerenje aktivnosti tripeptidil peptidaze 1 u homogenatu leukocita
Title (english)Fluorimetric method for measuring activity of the tripeptidyl peptidase 1 in the leukocyte homogenate
AuthorNikolina Blažević
MentorKsenija Fumić (mentor)
Committee memberKsenija Fumić (predsjednik povjerenstva)
Committee memberPetra Turčić (član povjerenstva)
Committee memberMarija Grdić Rajković (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Pharmacy and Biochemistry
(Department of medical biochemistry and haematology)
Zagreb
Defense date and country2019-07-08, Croatia
Scientific / art field,
discipline and subdiscipline		BIOMEDICINE AND HEALTHCARE
Pharmacy
Medical Biochemistry
Abstract
Neuronalne ceroidne lipofuscinoze (NCL) su nasljedne, progresivno degenerativne bolesti koje prvenstveno utječu na mozak i mrežnicu. One se smatraju lizosomalnim poremećajima skladištenja (LSD), zbog njihovog karakterističnog nakupljanja autofluorescentnih ceroidnih lipopigmenata, podjedinica C mitohondrijskih ATP sintetaza ili sfingolipidnih aktivatorskih proteina A i D u lizosomima većine stanica. Klasični kasni infantilni oblik NCL je uzrokovan mutacijama u CLN2 genu koji kodira lizosomalnu tripeptidil peptidazu 1. Klasični kasni infantilni fenotip CLN2 bolesti ima predvidljiv klinički tijek obilježen epilepsijom i brzim padom psihomotoričkih sposobnosti. Najčešći početni simptomi su kašnjenje razvoja govora i napadaji, koji se obično počinju pojavljivati u dobi između 2 i 4 godine. Rana dijagnoza CLN2 (neuronalna ceroidna lipofuscinoza tipa 2) bolesti je presudna kako bi se osigurala optimalna skrb za bolesnike i njihove obitelji. CLN2 bolest je moguće dijagnosticirati mjerenjem aktivnosti enzima TPP1. TPP1 enzim je pepstatin-nestabilna lizosomska serin egzopeptidaza s optimalnom in vitro aktivnošću u kiselom pH. Razvijena je rekombinantna terapija za liječenje CLN2, cerliponaza alfa, koja je najučinkovitija kod primjene nakon pojave prvih simptoma bolesti. Cilj rada je uvođenje metode za mjerenje aktivnosti enzima tripeptidil peptidaze 1 u uzorku leukocita izoliranih iz pune krvi. In house fluorimetrijska metoda određivanja aktivnosti enzima tripeptidil peptidaze 1 u leukocitima izoliranim iz EDTA krvi pokazala se kao pouzdana metoda za postavljanje dijagnoze CLN2. Takva se metoda može koristiti u selektivnoj laboratorijskoj dijagnostici nakon postavljene kliničke sumnje na CLN2. Za primjenu u rutinskoj laboratorijskoj dijagnostici potrebno je proširiti ispitivanje referentnog intervala na većem broju uzoraka, prvenstveno djece do pete godine života, jer su oni ciljana populacija za buduću CLN2 dijagnostiku. Međutim, zbog brojnih ograničenja fluorimetrijske metode, kao prva metoda probira preporučuje se metoda mjerenja aktivnosti TPP1 iz uzorka suhe kapi krvi na filter papiru.
Abstract (english)
The neuronal ceroid lipofuscinoses (NCLs) are inherited progressive degenerative diseases that primarily affect the brain and retina. They are considered lysosomal storage disorders (LSD), because of their characteristic accumulation of autofluorescent ceroid lipopigments, subunit c of mitochondrial ATP synthase or sphingolipid activator proteins A and D in lysosomes of most cells. The classic late infantile onset NCL is caused by mutations in CLN2 gene which encodes lysosomal tripeptidyl peptidase 1. The classic late-infantile phenotype of CLN2 disease has a predictable clinical course marked by epilepsy and rapid psychomotor decline. The most common initial symptoms are language delay and seizures, which typically begin to manifest between the ages of two to four years. Early diagnosis of CLN2 disease is crucial to ensure optimum care for patients and their families. CLN2 disease can be diagnosed by measuring the activity of the enzyme TPP1. TPP1 enzyme is a pepstatin-insensitive lysosomal serine exopeptidase with optimal in vitro activity at acidic pH. Recombinant therapy for the treatment of CLN2 has been developed, cerliponase alpha, which is most effective when is applied after the first symptoms of the disease. The aim of the study was to introduce methods for measuring the activity of the enzyme tripeptidyl peptidase 1 in a sample of leukocytes isolated from full blood. In house fluorimetric method of determining the activity od the enzyme tripeptidyl peptidase 1 in leukocytes isolated from EDTA blood has proven to be a reliable method for diagnosing CLN2. Such a method can be used in selective laboratory diagnostics after clinical suspected CLN2. For routine laboratory diagnostics, it is necessary to extend the reference interval test on a large number of samples, primarily children up to the age of five, as they are a target population for future CLN2 diagnostics. However, due to the numerous limitations of the fluorimetric method, the method of measurement of TPP1 activity from the dry blood sample on filter paper is recommended as the first method of screening.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:163:125241
Study programmeTitle: 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 resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2020-02-14 14:18:44