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Synthesis and characterization of peptide-functionalized noble metal nanoparticles and nanoclusters and their antibiotic activity
No full textNanostrukture, primjerice nanočestice i nanoklasteri, mogu pokazivati citotoksičnu aktivnost ali i sposobnost da djeluju kao nosači određenih molekula (lijekovi, proteini...). U ovom se diplomskom radu provela sinteza nanočestica i nanoklastera srebra i zlata nakon čega su nanostrukture povezane s antimikrobnim peptidima. Antimikrobni peptidi su klasa kratkih peptida koji se mogu naći u prirodi, a imaju širok raspon inhibicijskih učinaka prema bakterijama i virusima. Koristili su se indolicidin, anoplin i PMAP-23. Cilj rada je bio da se njihovim povezivanjem ispita postojanje potencijalnog sinergijskog efekta koji dolazi od nanostruktura i antimikrobnih peptida zajedno. Povezivanjem nanostruktura s antimikrobnim peptidima dobile su se funkcionalizirane nanostrukture, čiji se antibiotski potencijal provjeravao stavljanjem na hranjivu podlogu s bakterijom Escherichia coli. Karakterizacija prekursora i produkata sinteze odvijala se različitim analitičkim metodama. Konjugat nanočestica-peptid se okarakterizirao fluorescencijskom spektrometrijom i dinamičkim raspršenjem svjetlosti. Izgled (oblik, struktura) i veličina nanostruktura se provjeravala elektronskom mikroskopijom i putem UV-Vis spektrofotometrije. Antimikrobni peptidi su karakterizirani cirkularnim dikroizmom i UV-Vis spektrofotometrijom.Nanostructures, for example, nanoparticles and nanoclusters, can show cytotoxic activity and the ability to act as carriers of certain molecules (medicines, proteins...). In this thesis, nanoparticles and nanoclusters of silver and gold were synthesised, after which the nanostructures were conjugated with antimicrobial peptides. Antimicrobial peptides are a class of short peptides found in nature and have a wide range of inhibitory effects against bacteria and viruses. Indolicidin, anoplin and PMAP-23 were used. The goal was to examine the potential synergistic effect that comes from nanostructures and antimicrobial peptides together by connecting them. Functionalized nanostructures were obtained by linking nanostructures with antimicrobial peptides, whose antibiotic potential was determined by placing them on a nutrient medium with Escherichia coli. Characterization of precursors and synthesis products was carried out using various analytical methods. The nanoparticle-peptide conjugate was characterized by fluorescence spectrometry and dynamic light scattering. The appearance (shape, structure) and size of the nanostructures were determined by electron microscopy and UV-Vis spectrophotometry. Antimicrobial peptides were characterised by circular dichroism and UV-Vis spectrophotometry
Investigation of local structure in complex oxides of titanium and tantalum using neutron Larmor precession
No full textU ovom radu predstavljeni su rezultati Larmorove difrakcije neutrona na dva glavna predstavnika kvantnih paraelektrika, stroncijevom titanatu i kalijevom tantalatu. Istraženi su nedopirani uzorci, kao i uzorci dopirani niobijem i vakancijama kisika. Promatranjem promjene razmaka rešetke utvrđeno je da negativna termalna ekspanzija nedopiranog stroncijevog titanata nestaje pri dopiranju. Ovaj rezultat implicira snažan utjecaj dopiranja na anharmoničnost fonona u stroncijevom titanatu. Isti rezultat nije pronađen u kalijevom tantalatu, gdje je termalna ekspanzija pozitivna i u nedopiranom i dopiranom uzorku. Također, određene su i distribucije razmaka rešetke stroncijevog titanata, u kojima je uočeno cijepanje na dvije srednje vrijednosti razmaka rešetke, što je u skladu s poznatim antiferodistortivnim faznim prijelazom. Iz distribucija je moguće odrediti međusobni omjer zastupljenosti vrijednosti razmaka u uzorku, kao i širinu distribucije koja daje ideju o unutarnjoj deformaciji uzoraka. Dobivena vrijednost unutarnje deformacije u nedopiranom stroncijevom titanatu je veća od kritične vrijednosti elastične deformacije koja inducira feroelektričnost, što može objasniti formaciju polarnih nanoregija pri niskim temperaturama.representative quantum paraelectrics: strontium titanate and potassium tantalate. Both undoped samples and samples doped with niobium and oxygen vacancies were investigated. By looking into the change of lattice-spacings, we discover that the negative thermal expansion of strontium titanate vanishes with very low doping. This result implies that doping has a strong effect on phonon anharmonicity in strontium titanate. The same result is not found in potassium tantalate, where thermal expansion is positive in both the undoped and doped samples. Additionally, distributions of lattice-spacings of strontium titanate are determined for various temperatures and a splitting of distributions into two mean values is observed. This is in agreement with the known antiferrodistortive phase transition. From these distributions, it is possible to determine the ratio of the mean value populations, as well as the width of the distribution which provides an idea of intrinsic strain. The obtained intrinsic strain values in undoped strontium titanate are above the critical value of elastic strain that induces ferroelectricity, which can explain the formation of polar nanoregions at low temperatures
Synthesis and separation properties of zirconium based homochiral metal-organic frameworks with amino acid derived ligands
No full textU okviru rada je iz fumarne kiseline u 4 koraka sintetizirana ciklobutan-1,2,3,4-tetrakarboksilna
kiselina (cBTA) na koju su zatim amidnom vezom vezane po 4 molekule različitih
aminokiselina: L-alanina, L-fenilalanina i L-prolina. Dobiveni homokiralni ligandi korišteni su
za pripravu metaloorganskih mreža (MOF-ova) u solvotermalnim i mehanokemijskim
reakcijama sa spojevima cirkonija(IV). Separacijska svojstva MOF-a izvedenog iz L-alanina
ispitana su pomoću tri kiralna organska spoja. Katalitička svojstva istog MOF-a ispitana su na
aldolnoj reakciji cikloheksanona i 4-nitrobenzaldehida. Separacija je praćena tekućinskom
kromatografijom visoke djelotvornosti s kiralnom stacionarnom fazom. Sintetizirani ligandi
analizirani su spektroskopijom NMR, masenom spektrometrijom i IR spektroskopijom. MOFovi su analizirani difrakcijom rentgenskog zračenja na polikristalnom uzorku,
termogravimetrijom, IR spektroskopijom te spektroskopijom NMR u čvrstom stanju.Within this thesis cyclobutane-1,2,3,4-tetracarboxylic acid (cBTA) was synthesised in 4 steps
starting from fumaric acid. Amide bonds were formed between cBTA and 4 molecules of
different amino acids: L-alanine, L-phenylalanine and L-proline. Obtained homochiral ligands
were used to prepare metal-organic frameworks (MOFs) through solvothermal and
mechanochemical reactions with zirconium(IV) compounds. Separation properties of the MOF
derived from L-alanine were investigated using different chiral organic compounds followed by
high performance liquid chromatography with a chiral stationary phase. Catalytic properties of
the MOF were investigated through the aldol reaction of cyclohexanone and
4-nitrobenzaldehyde. Synthesised ligands were analysed using NMR spectroscopy, mass
spectrometry and IR spectroscopy. MOFs were analysed using powder x-ray diffraction,
thermogravimetry, IR and solid-state NMR spectroscopy
FILOGENETSKI ODNOSI UNUTAR RODA Phoxinus (TELEOSTEI: CYPRINIDAE) NA PODRUČJU ZAPADNOG BALKANA
No full textMinnows (genus Phoxinus) are small, cyprinid species of fish, widely distributed
throughout Europe and Asia, including in Croatia and neighboring countries. Currently, the
genus Phoxinus has as many as 23 different mitochondrial genetic lineages, thirteen
described species, while three species have been named, but their status has not yet been
confirmed. Until a few years ago, it was thought that two species were widespread in the
Western Balkans: Eurasian minnow (Phoxinus phoxinus) and the Adriatic minnow (P.
lumaireul). The first investigations showed the absence of the P. phoxinus species in this
area and the exceptional diversity of the P. lumaireul species with the existence of new
genetic lines and undescribed species. The aim of this doctoral investigation is to provide a
more detailed and clearer insight into the phylogenetic and phylogeographic as well as interand
intra-species relationships within the genus Phoxinus by a combination of molecular
phylogenetics and population genetics methods. The research showed a wide distribution of
the species P. lumaireul and a great genetic diversity in the Western Balkans, with the
existence of four genetic subclades in the area of Croatia and Slovenia. Two new species of
the Phoxinus genus were discovered in Croatia, one of which is new to science. By
combining the methods of molecular phylogenetics with morphology and meristics, this
species was described as Phoxinus krkae. For the needs of more detailed research on
population genetics within the genus, but especially within the P. lumaireul s.l. complex, a
new set of microsatellite markers was developed, which will provide detailed insights into
mutual population relationships, hybridization events and natural adn human-mediated
introgressions.Pijori (rod Phoxinus) su male, ciprinidne vrste riba, široko rasprostranjenje na
području cijele Europe i Azije, pa tako i na području Hrvatske i susjednih zemalja. Trenutno
rod Phoxinus broji čak 23 različite mitohondijske genetičke linije, trinaest opisanih vrsta,
dok je trima vrstama dodijeljeno ime, ali njihov status još nije potvrđen. Do prije nekoliko
godina se smatralo da su na području Zapadnog Balkana rasprostranjene dvije vrste: pijor
(Phoxinus phoxinus) i tzv. jadranski pijor (P. lumaireul). Prva istaživanja su pokazala
nepostojanje vrste P. phoxinus na ovom području te iznimnu raznolikost vrste P. lumaireul
uz postojanje novih genetičkih linija i neopisanih vrsta. Cilj ovog rada je pružiti detaljniji i
jasniji uvid u filogenetske i filogeografske te inter- i intra-specijske odnose unutar Phoxinus
roda kombinacijom raznih molekularne filogenije i populacijske genetike. Istraživanje je
pokazalo široku rasprostranjenost vrste P. lumaireul te veliku genetsku raznolikost na
području Zapadnog Balkana, uz postojanje četiri genetičke podlinije na području Hrvatske i
Slovenije. Otkrivene su i dvije nove vrste roda Phoxinus na području Hrvatske, od kojih
jedna nova za znanost. Kombinacijom metoda molekularne filogenetike te morfologije i
meristike, ova vrsta je opisana kao Phoxinus krkae. Za potrebe detaljnijih istraživanja
populacijske genetike unutar roda, ali posebice unutar kompleksa P. lumaireul s.l., razvijen
je novi set mikrosatelitnih markera, koji će dati detaljne uvide u međusobne populacijske
odnose, hibridizacije te prirodne i ljudski uzrokovane introgresije
Influence of the way of preparation of zeolites on the strenght of their catalytic active sites
No full textPripravljena su dva tipa zeolitne strukture: beta i mordenit. Iako su zeoliti mikroporozni materijali, njihova aktivnost u katalitičkim reakcijama je limitirana internom difuzijom reaktanata i produkata, a ista je poboljšana uvođenjem hijerarhijske strukture. Dio uzoraka modificiran je tijekom sinteze dodatkom ”mezoporoznog” templata – cetiltrietilamonijevog bromida, dok je drugi dio postsintetski obrađen nagrizanjem pomoću vodene otopine natrijevog hidroksida, tetrapropilamonijevog hidroksida i klorovodičnom kiselinom. Ovisno o omjeru silicija i aluminija te vrsti zeolita birana je otopina ili smjesa otopina za nagrizanje točno određenih koncentracija. Katalitička aktivnost i efikasnost katalizatora može se poboljšati povećanjem broja i podešavanjem jakosti katalitički aktivnih mjesta (Brønstedovih i Lewisovih kiselih mjesta), što je postignuto postsintetskim uvođenjem odabranih dvovalentnih kationa metala postupkom ionska izmjene popračene termičkom obradom. Količina, položaj i jakost katalitički aktivnih mjesta u materijalu praćen je Fourier-transformiranom infracrvenom spektroskopijom u vakuumu pomoću desorpcije probne molekule, deuteriranog acetonitrila.Two types of zeolite structures were prepared: beta and mordenite. Although they are microporous materials by nature, intracrystalline diffusion of reactants and products of catalysis was limited, and is improved by introducing a hierarchical structure. Some samples were modified during synthesis by adding mesoporous template – cetyltrimethylammonium bromide, while the other samples was post–synthetically treated by etching using a solution of sodium hydroxide, tetrapropylammonium hydroxide, and hydrochloric acid. The etching solution or mixture of etching solutions was selected depending on the Si/Al ratio and zeolite topology. The catalytic activity and efficiency of the materials can be improved by increasing the number and adjusting the strength of catalytically active sites (Brønsted and Lewis acid sites), which can be achieved by ion–exchange with specific divalent metal cations. The number, position and strength of catalytically active sites was studied using vacuum Fourier-transform infrared spectroscopy of desorption of probe molecules, deuterated acetonitrile
The role of BPM1 protein in de novo DNA methylation mechanism during development of Arabidopsis thaliana L.
No full textProteini BPM iz vrste Arabidopsis thaliana primarnu ulogu ostvaruju u putu
proteasomalne razgradnje ovisne o ubikvitinu, u sklopu kojeg prepoznaju specifične supstrate
za razgradnju. Prethodno je utvrđena interakcija proteina BPM1 s proteinima RDM1 i DMS3,
komponentama metilacijskog mehanizma RdDM. Uloga proteina BPM1 u mehanizmu RdDM
u ovom je radu ispitana primjenom kromatinske imunoprecipitacije u svrhu identifikacije novih
ciljnih gena ovog mehanizma u čijoj regulaciji metilacije posreduje protein BPM1.
Identificirani su geni FBW2, RKP i AGL14, a kao referentni odabran je gen CML41. Analizom
metilacije ovih gena metodom bisulfitnog sekvenciranja i pirosekvenciranja u linijama s
prekomjernom ekspresijom proteina od interesa (oeBPM1, oeDMS3), te linijama s narušenom
funkcijom mehanizma RdDM (dms3-1, nrpe1-10), potvrđen je stimulativni učinak proteina
BPM1 na aktivnost mehanizma, koji je najizraženiji tijekom embriogeneze, a sam mehanizam
RdDM značajniji je za zigotnu nego za somatsku embrigenezu. Ovisno o poziciji, učinci
metilacijskih događaja reflektiraju se na ekspresiju svih analiziranih gena.Proteins BPM of Arabidopsis thaliana have a primary role in the ubiquitin-dependent
proteasomal degradation pathway, where they recognize specific substrates for degradation.
The interaction of BPM1 protein with RDM1 and DMS3, components of RdDM methylation
mechanism, has previously been established. Here, the role of BPM1 protein in RdDM was
analyzed using chromatin immunoprecipitation for identification of new RdDM targets whose
methylation is mediated by BPM1 protein. Genes FBW2, RKP and AGL14 were identified as
targets, and CML41 was chosen as a reference gene. Methylation analysis of these genes was
conducted using bisulfite sequencing and pyrosequencing in lines with overexpression of
BPM1 (oeBPM1) or DMS3 (oeDMS3), and lines with impaired RdDM function (dms3-1,
nrpe1-10). The results confirmed a stimulative role of BPM1 protein in RdDM. This
mechanism proved to be more significant in zygotic than somatic embyrogenesis. Depending
on the position, methylation had a diverse effect on expression of analyzed genes
Crystals of cadmium(II) polymers with halide and aminopyridine ligands: Preparation, characterization and mechanical properties ; Phase diagram - an example of good teaching practice
No full textOvaj diplomski rad sastoji se od dvije cjeline: istraživačkog i metodičkog dijela. U
istraživačkom dijelu rada ukupno je priređeno 5 novih spojeva kadmijevih(II) halogenida s 3-
aminopiridinskim ligandima [Cd(μ-Cl)(μ3-Cl)(3-NH2py)]n : [Cd(μ-Cl)2(3-NH2py)2]n (1a),
[Cd(μ-Br)2(μ-3-NH2py)2]n (2a), [CdI2(3-NH2py)2] (3), [CdCl2(3-NH2py)4]·EtOH (X = Cl, Br;
1b, 2b). Također su priređena tri literaturno poznata spoja s 4-aminopirdinskim ligandima
[CdX2(4-NH2py)2]n (X = Cl, Br, I; 4-6) u svrhu ispitivanje njihovih mehaničkih svojstava. Za
karakterizaciju priređenih spojeva korištena je difrakcija rendgenskog zračenja u polikristalnom
uzorku i infracrvena spektroskopija. Spojevima 1a, 1b, 2a, 2b i 3 određena je kristalna struktura
metodom difrakcije rendgenskog zračenja u jediničnom kristalu. Preliminarna ispitivanja
mehaničkog odziva provedena su na kristalima spojeva 2a, 3, 4, 5 koji su bili odgovarajuće
morfologije i zadovoljavajuće kvalitete.
U metodičkom dijelu rada načinjen je pregled i kritički osvrt stručne literature na temu Fazni
dijagram. Predložen je 90-minutni nastavni sat učenja otkrivanjem u kojemu su ciljana
obrazovna postignuća primjerena za uzrast učenika prvog razreda gimnazije te je detaljno
razrađen radni listić za učenike kao i uputa za nastavnika.This thesis consists of two parts: research and methodological parts. In the research part, five
new compounds of cadmium(II) halides with 3-aminopyridine ligands were prepared
[Cd(μ-Cl)(μ3-Cl)(3-NH2py)]n : [Cd(μ-Cl)2(3-NH2py)2]n (1a), [Cd(μ-Br)2(μ-3-NH2py)2]n (2a),
[CdI2(3-NH2py)2] (3), [CdCl2(3-NH2py)4]·EtOH (X = Cl, Br; 1b, 2b), along with three
compounds with 4-aminopyridine ligands which are previously known from literature [CdX2(4-
NH2py)2]n (X = Cl, Br, I; 4-6). Diffraction of X-ray radiation in polycrystalline sample (PXRD)
and infrared spectroscopy (IR) were used to characterize the prepared compounds. The crystal
structure of compounds 1a, 1b, 2a, 2b and 3 was determined using single crystal X-ray
diffraction. Preliminary investigations of the mechanical properties of crystals were carried out
on crystals of compounds 2a, 3, 4, 5 with appropriate morphology and satisfactory quality.
In the methodological part of the paper, an overview and critical review of professional
literature on the topic Phase diagram was made. A 90-minute learning lesson has been proposed
by disclosure in which targeted educational achievements are appropriate for the age of first-
graders of the gymnasium and a worksheet for students as well as instructions for teacher have
been elaborated in detail
Impact of environmental enrichment on habituation period of the Common wall lizard (Podarcis muralis) in captivity
No full textPri proučavanju obrazaca ponašanja laboratorijskih životinja, uključujući habituaciju kao
jednostavan oblik učenja, često se koriste testovi istraživanja ponašanja. Svrha obogaćivanja
okoliša životinja u zatočeništvu jest podići kvalitetu života na višu razinu povećanjem interakcije
s okolinom. U slučaju održavanja gušterica u zatočeništvu dokazano je da vrlo važnu ulogu ima
kemosenzorno obogaćivanje. Za provedbu ovog istraživanja, ulovljeno je 20 ženki i 20 mužjaka
odraslih jedinki zidne gušterice (Podarcis muralis). Dvotjedni tretman kemosenzornog
obogaćivanja uključivao je prikupljanje mirisa jedinki pomoću destiliranom vodom namočenih
filter papira koji su se postavljali u skloništa za gušterice. Mirisi nepoznate gušterice su postavljani
u kutove terarija tretirane skupine, dok su se kontrolnoj skupini postavljali filter papiri bez mirisa.
U drugom tjednu tretmana sve jedinke podvrgnute su testu otvorenog polja u kojem je snimano
istraživačko ponašanje. Promatrane varijable, ukupna količina kretanja i učestalost palucanja
jezikom, analizirane su korištenjem računalnog programa EthoVision XT 15. Rezultati potvrđuju
bržu habituaciju tretiranih gušterica u odnosu na kontrolnu skupinu. Utvrđena je tendencija
postizanja uniformnijeg odgovora tretiranih jedinki. Smatram kako je vrlo važno laboratorijskim
istraživanjima doprinijeti novim spoznajama vezanim uz obogaćivanje okoliša gmazova i drugih
slabije istraživanih skupina organizama te mogućnostima njihove aktivne primjene.While studying behavior in laboratory animals, including habituation as a simple form of learning,
behavioral tests are often used. The purpose of environmental enrichment is to improve the quality
of life in captive animals by increasing interaction with surroundings. In the case of keeping lizards
in captivity, chemosensory enrichment plays very important role. For implementation of this
research, 20 female and 20 male adult common wall lizards (Podarcis muralis) were caught. The
two-week chemosensory enrichment treatment included individuals’ odor collection by using
distilled water-soaked filter papers placed in lizard shelters. Odors of unknown lizards were placed
in corners of the terrarium of experimental group, while odorless filter papers were placed in
control group. In the second week of treatment all individuals underwent an open field test in which
exploratory behavior was recorded. Observed variables, total amount of movement and frequency
of tongue flicking were analyzed using the EthoVision XT 15 software. The results confirm that
treated lizards habituated faster compared to the control group. A tendency to achieve more
uniform response of the treated individuals was determined. I believe it is very important to
contribute to new insights through laboratory research related to environmental enrichment of
reptiles and other less researched groups of organisms and the possibilities of their active
application
Quantitative determination of neonicotinoides and its metabolites in honeybees and honeybee products using liquid chromatography-tandem mass spectrometry
No full textRazvijena je metoda za kvantitativno određivanje neonikotinoida i njihovih metabolita u
uzorcima pčela i pčelinjim proizvodima vezanim sustavom tekućinska kromatografija –
tandemna spektrometrija masa. Predloženi analitički postupci validirani su u skladu sa
smjernicama SANTE/12682/2019 ispitivanjem radnih značajki metode u svim ciljanim
matricama. Preciznost, izražena kao relativna standardna devijacija, bila je < 20 %, dok je
točnost, izražena kao analitički povrat bila u rasponu od 70 % do 120 %, za većinu
neonikotinoida i njihovih metabolita. U okviru validacije metoda procijenjena mjerna
nesigurnost iznosila je < 50 % za sve analite. Predloženom metodom analizirano je ukupno
664 uzoraka pčela i pčelinjih proizvoda uzorkovanih tijekom 2019. i 2020. godine u svim
regijama Republike Hrvatske. Ostatci neonikotinoida i njihovih metabolita detektirani su u
17,6 % uzoraka, a tiakloprid je bio najčešće prisutan analit u obje godine istraživanja.A method for the quantitative determination of neonicotinoids and their metabolites in
honeybee and bee product samples using the liquid chromatography - tandem mass
spectrometry system was developed. The proposed analytical procedures were validated in
accordance with the SANTE/12682/2019 guidelines by evaluating the performance
characteristics of the method in all target matrices. Precision, expressed as relative standard
deviation, was < 20 %, while accuracy, expressed as analytical recovery, ranged from 70 %
to 120 %, for most neonicotinoids and their metabolites. As part of method validation, the
estimated measurement uncertainty was < 50 % for all analytes. A total of 664 samples of
honeybees and bee products, sampled during 2019 and 2020 in all regions of the Republic of
Croatia, were analysed using the proposed method. Residues of neonicotinoids and their
metabolites were detected in 17.6 % of samples, and thiacloprid was the most frequently
present analyte in both years of the study
An application of settlement central functions in the calculation of the development index: the example of the Primorje-Gorski kotar County
No full textMjerenje regionalne razvijenosti nekog prostora važno je zbog njegovog kvalitetnog planiranja i usmjeravanja razvojnih politika. U tu svrhu, u Hrvatskoj se koristi indeks razvijenosti, kompozitni pokazatelj koji u sebi sadrži skup demografskih i ekonomskih varijabli. No, pregledom relevantne literature utvrđeno je da sadrži određene nedostatke, ponajviše u metodološkom smislu. Zbog toga se otvorila mogućnost njegova unaprjeđenja pomoću novog pokazatelja – indeksa razvijenosti centralnih funkcija. Naime, centralne se funkcije u geografiji uvelike koriste kako bi se njima predočila razvijenost određenog prostora, te kao takve predstavljaju kvalitetnu osnovu za nadgradnju ovog pokazatelja. Stoga su oba pokazatelja metodom ranga stavljeni u međuodnos te je posljedično konstruiran kombinirani pokazatelj koji je objedinio oba navedena. Na primjeru Primorsko-goranske županije pokazalo se da on značajno ublažava izražene razlike u poretku jedinica lokalne samouprave prema razvijenosti, te da smanjuje dotad vidljivu polarizaciju u prostoru. Na posljetku, pokazalo se da novi, kombinirani pokazatelj ima potencijala za primjenu i korištenje u praksi.Measuring the regional development of an area is important because of its quality planning and direction of development policies. For this purpose, the Republic of Croatia uses the development index, a composite indicator that contains a set of demographic and economic variables. But, a review of the relevant literature found that it contains certain shortcomings, mostly in the methodological sense. Because of this, the possibility of its improvement using a new indicator - the index of the development of central functions has opened up. Namely, the central functions in geography are widely used in order to represent the development stage of a certain area, and as such they are a quality basis for the superstructure of this indicator. Therefore, both indicators were interrelated using the rank method, and as a result, a combined indicator was constructed that united both of them. On the example of the Primorje-Gorski Kotar County, it was shown that it significantly reduces the expressed differences in the order of local self-government units according to development, and that it reduces the previously visible polarization in space. In the end, it was shown that the combined indicator has potential for application and use in practice