Repository of the University of Rijeka, Department of Biotechnology
Not a member yet
715 research outputs found
Sort by
Sinteza peptida na čvrstom nosaču
No full textPeptides are biomolecules composed of two to fifty amino acids, connected into chains through peptide bonds. They have a wide variety of applications in medicine, biology, chemistry and other biomedical-related fields because of their antimicrobial, antiviral and catalytic activities. Solid phase peptide synthesis (SPPS) is the method used for producing these compounds in the laboratory. It consists of a few basic steps, namely: deprotection, coupling, washing, acetylation, and cleavage.
This undergraduate thesis is focused on the Fmoc SPPS synthesis, analysis, and purification of the Ac-HREAKRTRVCYR-Am (Ac=acyl group; Am=amide group) peptide and provides summarized protocols for all used methods.
During the synthesis, difficulties regarding the removal of the protecting group and liquid chromatography-mass spectrometry analysis were encountered due to the high hydrophilicity of the peptide. For this reason, the aim was to find different ways to optimize the method for synthesis, characterization, and purification to achieve a peptide of high purity that could undergo different biocatalytic assays in the future.Peptidi su biomolekule sastavljene od 2 do 50 aminokiselina povezanih u lanac peptidnim vezama. Imaju široku primjenu u medicini, biologiji, kemiji i ostalim srodnim biomedicinskim područjima zbog antimikrobnih, antivirusnih i katalitičkih sposobnosti. Sinteza peptida na čvrstom nosaču (engl. solid phase peptide synthesis; SPPS) metoda je koja se koristi za proizvodnju ovih spojeva u laboratoriju. Sastoji se od nekoliko osnovnih koraka: uklanjanje zaštitne skupine s aminokiselina, spajanja aminokiselina u lanac (engl. coupling), ispiranja, acetilacije i skidanja peptida s nosača (engl. cleavage).
Ovaj završni rad usmjeren je na Fmoc SPPS sintezu, analizu i pročišćavanje peptida Ac-HREAKRTRVCYR-Am (Ac=acilna skupina; Am=amidna skupina) i pruža sažete protokole za sve korištene metode.
Tijekom sinteze pojavile su se poteškoće s uklanjanjem zaštitne skupine i analize tekućinskom kromatografijom zbog visoke hidrofilnosti peptida. Iz tog razloga, cilj je bio pronaći različite načine za optimizaciju metoda sinteze, karakterizacije i pročišćavanja kako bi dobili peptid visoke čistoće koji može biti podvrgnut različitim biokatalitičkim testovima u budućnost
Lijekovi protiv herpes simpleks virusa 1 i 2
No full textInfekcije herpes simpleks virusima (HSV) jedne su od najčešćih i
najraširenijih virusnih infekcija. Razlikujemo HSV-1 koji najčešće uzrokuje
orofaringealne infekcije i HSV-2 koji najčešće uzrokuje genitalne infekcije.
HSV su DNA virusi koji pripadaju virusnoj obitelji Herpesviridae.
Replikacijski ciklus HSV sastoji se od tri faze: faza produktivne infkekcije,
faza latencije te faza reaktivacije. Najčešće HSV infekcije nisu opasne i
brzo se liječe, ali u imunokompromitiranih osoba i novorođenčadi one
mogu dovesti do ozbiljnih komplikacija, a nerijetko i smrti. Komplikacije
koje se najčešće javljaju su keratitis koji potencijalno vodi do sljepoće,
encefalitis te brojne sistemske bolesti. Smrtnost neliječenih odraslih
pacijenata s encefalitisom je oko 70%, a novorođenčadi 80%. Velika
većina odraslih i novorođenčadi koja prebole encefalitis imaju trajna
neurološka oštećenja. Veliki problem predstavlja sve veća stopa zaraženih
HSV-2. Najčešće korišteni lijekovi za liječenje HSV infekcija su nukelozidni
analozi. Oni ciljaju virusnu DNA polimerazu kako bi spriječili replikaciju
virusa. Predvodnik ove skupine je aciklovir. Kako nukelozidni analozi
zahtijevaju aktivaciju putem virusne timidin kinaze (TK), vrlo često dolazi
do rezistencije na njih zbog mutacije u virusnoj TK. Zbog toga su razvijeni
lijekovi koji ne trebaju aktivaciju TK-om. Primjer takvog lijeka je cidofovir.
Uz nukelozidne analoge, postoje i analozi pirofosfata koji također ne
zahtjevaju fosforilaciju virusnom TK. Primjer ovakvog lijeka je foskarnet.
Novije skupine lijekova u razvoju ciljaju helikaza- primaza kompleks kako
bi zaustavili virusnu replikaciju. Drugi pristup su lijekovi koji djeluju na
sam ulazak virusa u stanicu. Unatoč brojnim naporima, do danas nije
pronađeno uspješno protu-HSV cjepivo. Iako je protuvirusna terapija
omogućila kontinuirano i značajno poboljšanje u liječenju primarnih i
ponovljenih infekcija, otpornost na trenutno dostupne lijekove i
dugotrajna toksičnost predstavljaju trenutnu i buduću prijetnju s kojom se
treba pozabaviti razvojem protuvirusnih spojeva usmjerenih na nove ciljne
molekuleHerpes simplex virus (HSV) infections are one of the most common and
widespread viral infections. There are two herpes simplex viruses: HSV-1,
which most often causes oropharyngeal infections, and HSV-2, which most
often causes genital infections. HSV are DNA viruses that belong to the
virus family Herpesviridae. The replication cycle of HSV consists of three
phases: the phase of productive infection, the phase of latency and the
phase of reactivation. The most common HSV infections are not
dangerous and can be treated quickly, but in case of immunocompromised
persons and newborns, serious complications, often even death, can
occur. Mortality of untreated adult patients with encephalitis is about
70%, and 80% for newborns. The vast majority of adults and newborns
who recover from encephalitis have permanent neurological damage. A
big problem is the increasing rate of HSV-2 infection. The most commonly
used drugs for the treatment of HSV infections are nucleoside analogues.
The leader of this group is acyclovir. As nucleoside analogues require
activation by the viral thymidine kinase (TK), resistance to them occurs
very often due to a mutation in the viral TK. For this reason, drugs that do
not need TK activation have been developed. An example of such a drug is
cidofovir. Foscarnet is a priphosphate analog. New classes of drugs in
development target the helicase-primase complex to stop viral replication.
Another approach is drugs that act on the entry of the virus into the cell
itself. Despite numerous efforts, no successful anti-HSV vaccine has been
found to date. Although antiviral therapy has enabled continuous and
significant improvement in the treatment of primary and recurrent
infections, resistance to currently available drugs and long-term toxicity
represent a current and future threat that needs to be addressed by the
development of antiviral compounds directed against new targets
Structural modifications introduced by NS2B cofactor binding to the NS3 protease of the Kyasanur forest disease virus
No full textKyasanur Forest Disease virus (KFDV), a neglected human pathogenic virus, is a Flavivirus that causes severe hemorrhagic fever in humans. To develop a new antiviral therapy against KFDV, we focused on the nonstructural proteins NS2B and NS3 of KFDV which are responsible for serine protease activity. The structure of the NS2B/NS3 protease of KFDV using AlphaFold was created and molecular dynamics (MD) simulations with and without NS2B cofactor were performed to investigate structural rearrangements due to cofactor binding and to identify alternative allosteric sites. 100 ns MD simulations for the top 4 hit molecules confirmed the stability of the allosteric complexes.Detailed protocol used for MD simulation is described in 'Structural modifications introduced by NS2B cofactor binding to the NS3 protease of the Kyasanur forest disease virus', Ticks and Tick-borne Disease
Gaussian field-based 3D-QSAR and molecular simulation studies to design potent pyrimidine–sulfonamide hybrids as selective BRAFV600E inhibitors
No full textThe “RAS-RAF-MEK-ERK” pathway is an important signaling pathway in melanoma. BRAFV600E(70–90%) is the most common mutation in this pathway. BRAF inhibitors have four types of conformers: type I (aC-IN/DFG-IN), type II (aC-IN/DFG-OUT), type I1/2(aC-OUT/DFG-IN), and type I/II (aC-OUT/DFG-OUT). First-and second-generation BRAF inhibitors show resistance to BRAFV600Eand are ineffective against malignancies induced by dimer BRAF mutants causing ‘paradoxical’ activation. In the present study, we performed molecular modeling of pyrimidine–sulfonamide hybrids inhibitors using 3D-QSAR, molecular docking, and molecular dynamics simulations. Previous reports reveal the importance of pyrimidine and sulfonamide moieties in the development of BRAFV600Einhibitors. Analysis of 3D-QSAR models provided novel pyrimidine sulfonamide hybrid BRAFV600Einhibitors. The designed compounds share similarities with several structural moieties present in first- and second-generation BRAF inhibitors. A total library of88 designed compounds was generated and molecular docking studies were performed with them. Four molecules (T109, T183, T160, and T126) were identified as hits and selected for detailed studies. Molecular dynamics simulations were performed at 900 ns and binding was calculated. Based on molecular docking and simulation studies, it was found that the designed compounds have better interactions with the core active site [the nucleotide (ADP or ATP) binding site, DFG motif, and the phospho-acceptor site (activation segment) of BRAFV600Eprotein than previous inhibitors. Similar to the FDA-approved BRAFV600Einhibitors the developed compounds have [aC-OUT/DFG-IN] conformation. Compounds T126, T160 and T183 interacted with DIF (Leu505), making them potentially useful againstBRAFV600Eresistance and malignancies induced by dimer BRAF mutants. The synthesis and biological evaluation of the designed molecules is in progress, which may lead to some potent BRAFV600Eselectiveinhibitors
Development of Raman spectroscopy as a noninvasive in-process control method in pharmaceutical industry : Master's Thesis
No full textKontinuirani proizvodni procesi farmaceutika sve su važniji zahvaljujući većoj efikasnosti, smanjenju troškova i boljim osiguranjem kvalitete u odnosu na serijsku proizvodnju. Kako bi kontinuirani proizvodni proces bio valjan nužno je osigurati primjerene metode za nadzor procesa u stvarnom vremenu u skladu s ICH Q2 (R1) smjernicama. Ramanova spektroskopija neinvazivna je vibracijska spektroskopska tehnika koja omogućava brzu kvantitativnu analizu nekog uzorka. Upravo iz tog razloga Raman uređaj opremljen sondom koja može biti implementirana u proizvodni proces obećavajući je način za praćenje sadržaja djelatne tvari (eng. active pharmaceutical ingreedient, API) u stvarnom vremenu. U provedenom istraživanju ispitana je prikladnost Ramanove spektroskopije kao metode za procesnu kontrolu (eng. in-process control, IPC) u proizvodnji tekućih farmaceutskih oblika s 1% w/w ili manje djelatne tvari. Napravljeni su Raman modeli za Meralys® sprej sadržaja ksilometazolin hidroklorida (Xylo HCl) 1 i 0.5 mg/ml te za Aknet® otopinu sadržaja klindamicin hidroklorida (Clinda HCl) 10 mg/ml. Izrađeni su univarijantni integracijski i hard modeli te multivarijantni modeli metodom parcijalnih najmanjih kvadrata (eng. partial least squares, PLS). Integracijski modeli nisu dali validacijske parametre unutar zahtjeva za niti jedan od API-ja. Za Xylo HCl najbolje rezultate dao je PLS model uz koeficijent korelacije (R2) od 0.999, pogrešku predviđanja (RMSEP) od 0,92% i razliku stvarne od predviđene vrijenosti sadržaja od -1,98 do +1,86%. Za Clinda HCl najbolje rezultate dao je hard model uz R2 od 0.997, RMSEP od 0,38% i razlikom od -0,70 do 0,98%. Niti jedan od razvijenih modela nije zadovoljio sve validacijske parametre. Konvencionalna Raman spektroskopija nije prikladna IPC metoda za praćenje sadržaja API-ja u odabranim proizvodima.Continuous manufacturing of pharmaceuticals becomes more important due to greater efficiency, cost reduction and better quality assurance compared to batch production. For the continuous manufacturing process to be valid it is necessary to provide adequate methods for real- time process monitoring in accordance with the ICH Q2 (R1) guidelines. Raman spectroscopy is a non-invasive vibrational spectroscopic technique that enables rapid quantitative characterization of analytes in a sample. For this reason, a conventional Raman device equipped with a probe implemented in the production process is a promising way to monitor the active pharmaceutical ingredient (API) content in real-time. In this research we investigated the suitability of Raman spectroscopy as an in-process control (IPC) method in the production of liquid pharmaceuticals with 1% w/w or less API. Raman models were created for Meralys® spray containing 1 and 0.5 mg/ml xylometazoline hydrochloride (Xylo HCl) and for Aknet® solution containing 10 mg/ml clindamycin hydrochloride (Clinda HCl). Univariate integration and hard models as well as multivariate partial least squares (PLS) models were created. The integration models did not provide validation parameters within the requirements for any of the APIs. For Xylo HCl, PLS model gave best results with a correlation coefficient (R2) of 0.999, a prediction error (RMSEP) of 0.92% and a difference between the real and the predicted API content from -1.98 to +1.86%. For Clinda HCl, hard model gave the best results with an R2 of 0.997, an RMSEP of 0.38% and the difference ranging from -0.70 to 0.98%. None of the developed models met all the validation parameters. Conventional Raman spectroscopy is not a suitable method for analyzing API content in selected products and as such cannot be accepted as IPC in accordance with ICH Q2 (R1) guidelines
Molekulsko dinamičke simulacije retinala u otopini
No full textSkup podataka, uglavnom kvantno-kemijskih računa (optimizacija, računa energija i simulacija UV i IR spektara), koji su rezultirali objavom tri rada:
1) Biliskov, N.; Novak, J.; Petkovic, M.; Zgrablic, G.; Baranovic, G.; Doslic, N., Localization of the Counterion of the Protonated Schiff Base of n-butylretinal in Solution, Croatica Chemica Acta, 2011, 84, 221-231
2) Malis, M.; Novak, J.; Zgrablic, G.; Parmigiani, F.; Doslic, .N, Phys. Chem. Chem. Phys., 2017, 19, 25970-25978
3) Novak, J.; Malis, M.; Doslic, N., Croatica Chemica Acta, 2020, 93, 303-31
miRNA REMODELING OF THE TUMOR MICROENVIRONMENT
No full textTumor je nakupina stanica koje imaju nekontroliran rast. Tumor nastaje tumorogenezom, složenim procesom pretvorbe normalnih stanica u tumorske u uvjetima hipoksije. Tumori se mogu podijeliti na dobroćudne (benigne) i zloćudne (maligne). Na proliferaciju i progresiju malignog tumora ne utječu samo tumorske stanice, nego i stanice koje se nalaze u tumorskom mikrookolišu (TME), a bitnu ulogu u razvoju imaju i microRNA (miRNA). Postoje dvije glavne podjele miRNA koje sudjeluju u metabolizmu tumora, onkomiR i anti-onkomiR. OnkomiR su miRNA koje mogu transformirati normalne stanice u tumorske, a anti-onkomiR mogu spriječiti transformaciju u tumorske stanice. miRNA se mogu smatrati ključnim elementom koji utječe na stanice TME jer sudjeluju u regulaciji genske ekspresije i na taj način mijenjaju TME i potiču lučenje citokina. Dugogodišnja istraživanja lijekova za tumore još uvijek nisu dala očekivane rezultate te su potrebna daljnja istraživanja i razumijevanje mehanizama koji pogoduju inicijaciji i progresiji tumora. Budući da je deregulacija miRNA povezana s razvojem i napredovanjem raka, cirkulirajuće miRNA razmatraju se kao novi, minimalno invazivni biomarkeri te su potencijalna terapeutska meta jer je dokazana mogućnost njihove manipulacije. Kako bi se miRNA mogle uspješno koristiti kao terapeutici potrebno je premostiti izazov isporuke miRNA u ciljana tkiva.A tumor is a mass of abnormal cells that have uncontrolled growth. The tumor is formed by tumorigenesis, a complex process of converting normal cells into tumor cells under conditions of hypoxia. Tumors can be divided into benign and malignant. The proliferation and progression of a malignant tumor is influenced not only by tumor cells, but also by cells located in the tumor microenvironment (TME), and microRNA (miRNA) also plays an important role in its development. There are two main divisions of miRNAs involved in tumor metabolism, oncomiR and anti-oncomiR. OncomiR are miRNAs that can transform normal cells into tumor cells, and anti-oncomiR can prevent transformation into tumor cells. miRNAs can be considered a key element that affects TMEs cells because they participate in the regulation of gene expression and thus alter TMEs and stimulate the secretion of cytokines. Many years of research into tumor drugs have not yet given the expected results, so further research and understanding of the mechanisms conducive to tumor initiation and progression is needed. Because deregulation of miRNAs is associated with cancer development and progression, circulating miRNAs are considered a new, minimally invasive biomarker and are a potential therapeutic target because the possibility for their manipulation has been proven. In order for miRNA to be used successfully as a therapeutic, it is necessary to bridge the challenge of delivering miRNA to target tissues
mRNA vaccines against COVID-19
No full textTeški akutni respiratorni sindrom koronavirus 2, virus je koji se pojavio
krajem 2019. godine u Wuhanu, proširio svijetom i uzrokovao pandemiju
koronavirusne bolesti 19. Zbog brzog širenja virusa i često
eksponencijalnog rasta zaraženih, javili su se brojni problemi u zdravstvu,
ali i svjetskoj ekonomiji i gospodarstvu. Kako bi se spriječilo širenje virusa
i smanjio broj pacijenata većina država uvodila je stroge epidemiološke
mjere te je zatvarala svoje granice. Pandemija COVID-19 zahtijevala je brzu
reakciju znanstvenika te su nekoliko dana nakon sekvenciranja genoma
virusa započela brojna istraživanja usmjerena razumijevanju biologije
virusa te terapijskim i preventivnim pristupima protiv COVID-19 bolesti.
Pritom su uspješno iskorištena prethodna znanja o cjepivima i modernoj
mRNA tehnologiji te je upravo mRNA cjepivo bilo prvo odobreno cjepivo
protiv koronavirusne bolesti. mRNA tehnologija u posljednjih deset godina
doživjela je velik razvoj te je danas često korištena tehnika u
eksperimentalnim pristupima liječenju raka. mRNA molekule za cjepiva se
sintetiziraju in vitro na temelju sekvence genoma ciljanog patogena te se
cijepljenjem unose u organizam nakon čega aktiviraju urođeni i stečeni
imunosni odgovor. Stoga se mRNA tehnologija ubraja u važne pristupe
prevencije COVID-19 bolesti te se očekuje da će pridonijeti završetku
pandemije COVID-19.Severe acute respiratory syndrome coronavirus 2, appeared in late 2019.
in Wuhan, spread around the world and caused coronavirus pandemic. Due
to rapid spread of the virus and almost an exponential growth of those
infected individuals, numerous problems arose in healthcare, as well as in
the world economy. In order to prevent spread of the virus and reduce the
number of patients, most states introduced strict epidemiological measures.
The COVID-19 pandemic required a quick response from scientists, so few
days after sequencing of the virus genome, intense research began to
understand the biology of the virus, and preventive and therapeutic
measures against COVID-19 infection. Previous knowledge of vaccines and
modern mRNA technology has been used and mRNA vaccine became first
approved vaccine against coronavirus disease. mRNA technology had a
major development in the last ten years and today it is often used technique
in experimental tumor targetting. mRNA molecules are synthesized in vitro
based on the pathogen genome sequence and are introduced into the body
by vaccination. After vaccination they stimulate a protective immune
response. mRNA technology is one of the major upcoming therapeutic
approaches and will likely contribute to ending COVID-19 pandemic
DNA metiltransferaza iz sisavaca DNMT1: razvoj novih inhibitora analizama strukture i funkcije enzima pomoću superračunala
No full textBackground: DNA methylation is considered a basic and longest-lasting mechanism of human genome organization. It is established during embryogenesis and lasts for the lifetime as the first step of epigenetic regulation. Dysregulation of DNA methylation causes apoptosis, aging, and various pathological conditions. The development of DNMT inhibitors is one of the greatest challenges for 21st-century pharmacology and biotechnology. DNA methyltransferases are a uniquely complex challenge considering the size and multiple complex regulatory mechanisms. Selective manipulation of DNA methylation in healthy tissues can be used to produce stem cells. In this study, we used a supercomputer to develop inhibitors and analyze the structure and function of the major human DNA methyltransferase DNMT1.
Results: The binding site of the SAM cofactor in the active site of the enzyme is evolutionarily highly conserved, indicating that this part of the enzyme is described the best. 4WXX, 3PTA, and 4DA4 are three structural forms of DNMT1 from the PDB database that show different degrees of enzyme activity regulation and subtle differences in the active site. DNMT1 showed the highest mobility in its RFTS, CXXC, and BAH domains. Different enzyme forms have been used to screen compounds from commercial small molecule libraries to find compounds with the best binding affinity for the DNMT1 active site. The compounds were initially selected by the docking method. By that, compounds that exhibit shape-complementarity with the DNMT1 active site and generate the largest number of interactions were identified. The best-ranked compounds were used to describe the detailed binding mechanism by MD over 100 ns. Binding modes of identified compounds were compared to the binding of cofactor SAH as a reference ligand with a well-known binding mechanism.
Conclusion: ELITE12 and MYB1 are two compounds from commercial compound libraries that do not show a significant improvement in stability and number of binding interactions relative to SAH. The development of small molecules with binding affinity to the DNMT1 active site depends on organic syntheses and specifically designed compounds.Pozadina: DNA metilacija je osnovni i najdugotrajniji mehanizam u funkcionalnoj organizaciji ljudskog genoma. DNA metilacija se uspostavlja tijekom embriogeneze i traje do kraja života kao prvi korak epigenetske regulacije. Poremećaji DNA metilacije uzrokuju apoptozu, starenje te različita patološka stanja. Razvoj DNMT inhibitora je jedan od najvećih komercijalnih izazova za farmakologiju i biotehnologiju 21. stoljeća. DNA metiltransferaze su jedinstveno složen izazov jer se radi o velikim enzimima s višestrukim složenim regulatornim mehanizmima. Selektivna manipulacija DNA metilacije u zdravim tkivima može se koristiti za proizvodnju matičnih stanica. U ovom istraživanju smo koristili superračunalo kako bismo razvili inhibitore te analizirali strukturu i funkciju glavne ljudske DNA metiltransferaze DNMT1.
Rezultati: Mjesto vezanja kofaktora SAM u aktivnom mjestu enzima je evolucijski visoko očuvano što ukazuje da taj dio enzima najbolje poznajemo. 4WXX, 3PTA i 4DA4 su tri strukturalne forme DNMT1 iz PDB baze podataka. One prikazuju različite stupnjeve regulacije aktivnosti enzima sa suptilnim razlikama u aktivnom mjestu. Enzimski oblici su pokazali najveću mobilnost DNMT1 u RFTS, CXXC i BAH domenama. Različiti oblici enzima su stoga korišteni za pregled spojeva iz komercijalnih baza malih molekula u potrazi za spojevima koji se mogu najbolje vezati u aktivno mjesto DNMT1. Spojevi su početno selektirani metodom docking-a, čime su indetificirani spojevi koji stvaraju najveći broj interakcija i svojim oblikom najbolje odgovaraju aktivnom mjestu DNMT1. Najbolji spojevi su se koristili za opisivanje detaljnog mehanizma vezanja molekularnom dinamikom kroz 100 nanosekundi. Vezanje identificiranih spojeva se uspoređivalo s vezanjem kofaktora SAH kao referentnog liganda koji ima dobro poznat mehanizam vezanja.
Zaključak: ELITE12 i MYB1 su dva spoja iz komercijalnih baza malih molekula koji ne pokazuju značajno poboljšanje u stabilnosti i broju veznih interakcija u odnosu na S-adenozil-homocistein. Razvoj malih molekula koje se vežu na aktivno mjesto DNMT1 ovisi o organskim sintezama i specifično dizajniranim spojevima
The role of dopamine in preferential self-administration of methamphetamine
No full textKonzumacija opojnih droga utječe na dopaminsku i serotoninsku neurotransmisiju te je značajan faktor razvoja ovisnosti. Psihostimulansi poput metamfetamina (METH) djeluju na centre u mozgu zadužene za regulaciju procesuiranja nagrade i ugode. Nagrađujući utjecaj droga dovodi do opetovane konzumacije i u konačnici do ovisničkog ponašanja koje se ispituje na laboratorijskim životinjama kao preferencijalna samoadministracija. Drosophila melanogaster poznati je modelni organizam za izučavanje ovisničkog ponašanja.
U provedenom istraživanju koristili smo fumin (fmn) mutante Drosopile melanogaster kao modelni organizam za ispitivanje preferencijalne konzumacije METH-a. Zanimalo nas je kakva će biti preferencijalna administracija u tih mušica koje zbog mutacije u genima za dopaminski transporter (DAT) imaju narušenu homeostazu u dopaminskom sustavu. Metoda pomoću koje se ispitivala preferencijalna konzumacija je Fly CAFE (eng. Capillary Feeder) esej u kojemu su mušice imale izbor između šećerne otopine i šećerne otopine s dodatkom METH-a.
Dobiveni rezultati upućuju da dopamin (DA) ima ključnu ulogu u preferencijalnoj konzumaciji METH-a. fmn mušice razvijaju odbojan do neutralan odgovor prema šećernoj otopini koja sadrži METH. Također prikazana je značajna statistička razlika u preferenci za METH-om između divljeg tipa (wt) mušica s normalnom funkcijom dopaminskog sustava i fmn s neaktivnim DAT-om.
Potrebna su daljnja istraživanja i razumijevanje samoadministracije opojnih droga i njihovog nagrađujućeg djelovanja kako bi se dizajnirali prikladni farmakološki tretmani u liječenju i sprječavanju ovisnosti.The consumption of narcotic drugs affects dopamine and serotonin neurotransmission and is a significant factor in the development of addiction. Psychostimulants such as methamphetamine (METH) act on centers in the brain responsible for regulating reward and pleasure processing. The rewarding influence of drugs leads to repeated consumption and ultimately to addictive behavior that is tested on laboratory animals as preferential self-administration. Drosophila melanogaster is a well-known model organism for studying addictive behavior.
In the conducted experiment we used fumin (fmn) mutants of Drosopila melanogaster as a model organism for testing the preferential consumption of METH. We were interested to test the preferential administration in flies that, due to a mutation in the genes for the dopamine transporter (DAT), have disturbed homeostasis in the dopamine system. The method used to test preferential consumption is the Fly CAFE (Capillary Feeder) assay in which the flies had a choice between a sugar solution and a sugar solution with the addition of METH.
We found that dopamine plays a key role in the preferential consumption of METH. fmn flies develop an aversive to neutral response to a sugar solution containing METH. A significant statistical difference in preference for METH was also shown between wild type flies (wt) with normal dopamine system function and fmn with with an inactive DAT.
Further research and understanding of self-administration of narcotic drugs and their rewarding effects will be useful in design of appropriate pharmacological treatments for the treatment and prevention of addiction