Capacity Building for Rare Genomics

Infrequent occurrence of rare diseases (RD) has led to prolonged time needed to make an accurate diagnosis. Research and development of new treatments has been insufficient due to the narrow market for affected patients. Fascinating progress and accumulation of knowledge in the field of genomics provided for improvement of diagnosis and follow-up of numerous RD. It also led to the design of molecular-targeted therapeutics and more effective treatment of RD. Building up a centre for genomics of rare diseases comprises strengthening of several components in an institution already involved in molecular genetics and biomedical research, such as, infrastructure, human resources, local and international networking and dissemination and outreach activities. Regarding infrastructure, facilities for Next-generation sequencing (NGS) and array-based comparative genomic hybridization (aCGH) are necessary to perform high-throughput genomewide screening. Establishment of the bioinformatics unit is also very important. At the heart of genomics of RD is biological samples storage and database facilities. Biobanks are a critical resource in the field of genomics of RD. Additionally, strong connections with local medical institutions have to be made. Reinforcement of human potential, especially through recruitment of experts in bioinformatics, is indispensable contribution to the implementation of new equipment and high-throughput methodologies. Trainings of researchers are continually in focus of an expert centre for genomics of RD. Integration of the centre into European networks in order to intensify the transfer of knowledge is one of the cornerstones for its capacity building. The data a nd biospecimens that are distributed to all researchers, are absolute requirements in the pathway to develop modern science, diagnosis and cures for RD. Finally, dissemination and outreach activities are needed to ensure increased visibility of the centre within local and international scientific community and stakeholders and social entities interested in exploiting the results of the centre

Molecular diagnosis of phenylketonuria: From defective protein to disease-causing gene mutation

Fenilketonurija (PKU) najčešći je urođeni metabolički poremećaj u populaciji belaca (1/10000). Fenilketonurija nastaje kao posledica više od 500 mutacija u genu za fenilalanin hidroksilazu (PAH) koje dovode do deficijencije u aktivnosti enzima fenilalanin hidroksilaze (PAH). Pomoću in vitro ekspresione analize mutiranog enzima PAH i genotip-fenotip korelacije procenjuje se težina mutacija. Pokazano je da postoji značajna korelacija između genotipova u kojima su prisutni mutirani aleligena PAH i fenotipa PKU. Zbog toga je detekcija mutacija u genu za fenilalanin hidroksilazu sastavni deo moderne dijagnostike PKU. Studija molekularne osnove fenilketonu rije u Srbiji obuhvatila je identifikaciju spektra i frekvencije mutacija PAH i analizu korelacije genotipa i fenotipa pacijenata. Kombinovanjem metoda, PCR-RFLP, DGGE širokog spektra i DNK sekvenciranja, postignut je nivo detekcije mutacija od 97%. Na taj način je u Srbiji postavljena osnova za molekularnu dijagnostiku, genetsko savetovanje i odabir pacijenata sa fenilketonurijom kojima bi BH4 terapija bila od koristi.Phenylketonuria (PKU) is the most common inborn error of amino acid metabolism, with an average incidence of 1/10000 in Caucasians. PKU is caused by more than 500 mutations in the phenylalanine hydroxylase gene (PAH) which result in phenylalanine hydroxylase (PAH) enzyme deficiency. Two approaches, in vitro expression analysis of mutant PAH and genotypephenotype correlation study, are used for the assessment of severity of PAH mutations. It has been shown that there is a significant correlation between mutant PAH genotypes and PKU phenotypes. As a result, the molecular diagnosis is completely shifted toward the detection of mutations in the phenylalanine hydroxylase gene. The study of the molecular basis of PKU in Serbia included identification of the spectrum and frequency of PAH mutations in Serbian PKU patients and genotype-phenotype correlation analysis. By using both PCR-RFLP and 'broad range' DGGE/DNA sequencing analysis, the mutation detection rate reached 97%. Thus, the base for molecular diagnosis, genetic counseling and selection of BH4-responsive PKU patients in Serbia was created

Хибридни дуо

As members of the LP Duo, for the past 14 years we have experimented with various possibilities of playing on two pianos. Our artistic curiosity and the desire to gain new knowledge and freedom led to our involvement with the Quantum Music project and the creation of a new instrument – the hybrid piano. In this paper we elaborate on our experience with using the new hybrid pianos within the Quantum Music project, but also discuss our ensemble as an artistic embodiment of duality, entanglement and other quantum phenomena. Hybrid piano is a combination of a traditional piano and digital synthesizer with analogue controls that allows diferent processing of acoustic and digital signals in real time and enables the pianists to continue to use concert pianos whilst equipping them with new colours and new, hitherto unexlplored expressive possibilities.Београдски клавирски дуо Соња Лончар и Андрија Павловић (ЛП Дуо) већ четрнаест година експериментише са различитим могућностима свирања на два клавира. Поред класичног репертоара који је овај дуо прошао за време и након oсновних и специјалистичких студија на ФМУ у Београду а потом и на Високој школи за музику и позориште у Роштоку (Немачка), његови чланови су од самог почетка испољавали интересовање за савремену уметност, извођење дела композитора-савременика, бављење визуелним и примењеним уметностима (видео, филм, фотографија, музички перформанс, кореографије, позоришне представе), те свирање у ансамблима и музичким саставима различитих жанрова (рок, популарна, експериментална музика), као и свирање на различитим синтисајзерима. Њихов истраживачки дух и потреба за освајањем нових знања и слобода, као и сва наведена искуства, постепено су водила ка осмишљавању и реализацији пројекта Квантна музика и стварању новог инструмента – хибридног клавира. У овом раду чланови ЛП Дуа преносе своја искуства рада на пројекту Квантна музика и на развоју хибридног клавира, док у исто време свој ансамбл посматрају као отелотворење квантних феномена попут дуалности. Хибридни клавир је комбинација традиционалног клавира и дигиталног синтисајзера са аналогним контролама које омогућавају различито процесовање акустичких и дигиталних сигнала у реалном времену и омогућавају пијанистима да наставе да користе концертне клавире, али обогаћене новим бојама и досад незамисливим изражајним могућностима.Some parts of this article were incorporated into authors' (unpublished) doctoral dissertations: Lončar, Sonja (2018) „Hibridni duo – Novi prostori savremenog izvođaštva u ansamblu klavirski duo.“ [Hybrid Duo – New realms of contemporary performance for the piano duo.] Unpublished dissertation. Belgrade: Faculty of Music, University of Arts; Pavlović, Andrija (2018) „Savremeni izazovi rada i saradnje muzičara u klavirskom duu na primeru muzičko-scenskog projekta Kvantna muzika.“ [Contemporary challenges of work and collaboration between musicians in a piano duo, using the example of the music-scenic project Quantum Music.] Unpublished dissertation. Belgrade: Faculty of Music, University of Arts

Individualized therapy: Role of thiopurine S-methyltransferase protein and genetic variants

Tiopurin S-metiltransferaza (TPMT: EC 2.1.1.67) jeste enzim koji metaboliše imunosupresivne tiopurinske lekove, koji se koriste za lečenje autoimunih bolesti, malignih oboljenja i u transplantacionoj medicini. Aktivnost enzima TPMT kod pojedinih ljudi je izrazito smanjena ili povećana u odnosu na normalni nivo aktivnosti. Istraživanja strukture i biohemijskih karakteristika proteina TPMT su ukazala na postojanje određenih proteinskih varijanti koje imaju izmenjenu aktivnost. Otkriveni su polimorfizmi u genu za TPMT koji daju različite TPMT alozime. Smanjenoj aktivnosti enzima može doprineti i manja količina sintetisanog proteina, što zavisi i od transkripcione aktivnosti promotora gena za TPMT. Polimorfizmi u samom promotoru, kao što je promenjiv broj tandemskih ponovaka (VNTR), mogu da modulišu transkripciju. Primena tiopurinskih lekova kod pacijenata sa određenim genetskim varijantama TPMT izaziva tešku hematološku toksičnost. Da bi se toksičnost izbegla, terapija se modifikuje u skladu sa genotipom TPMT (farmakogenetika). Mi smo izučavali polimorfizme u egzonima i regulatornim elementima (promotor) gena za TPMT koji dovode do promene aktivnosti enzima TPMT u srpskoj populaciji. Koristili smo metodologiju baziranu na PCR i DNK sekvenciranje za detekciju genetskih varijanti TPMT. Pokazali smo da su u našoj populaciji prisutne genetske varijante u egzonima koje ukupno daju 7,5% varijantnih alozima TPMT koji imaju smanjenu enzimsku aktivnost. Terapija za pacijente koji imaju ove farmakogenetičke markere je modifikovana, što je doprinelo uspešnijem lečenju. Funkcionalnim esejima in vitro smo pokazali da aktivnost promotora gena za TPMT, a samim tim i količina sintetisanog enzima TPMT, zavisi od arhitekture (broja i tipa) VNTR u promotoru. Promotor gena za TPMT specifično odgovara na tretman ćelija K562 tiopurinom zavisno od tipa VNTR. Izučavanje interakcija DNK i proteina je otkrilo da transkripcioni faktori Sp1 i Sp3 interaguju sa VNTR. Naša istraživanja ukazuju na to da bi region VNTR u promotoru gena za TPMT mogao postati novi farmakogenetički marker od kliničkog značaja za individualizaciju tiopurinske terapije.Thiopurine S-methyltransferase (TPMT: EC 2.1.1.67) is an enzyme that metabolizes immunosuppressive thiopurine medications, used in the treatment of autoimmune diseases, cancer and in transplantation medicine. In some individuals, TPMT enzyme activity is significantly increased or decreased compared to the normal TPMT activity level. Structural and biochemical analyses of the TPMT protein revealed the existence of certain protein variants with altered activity. It has been shown that certain TPMT gene polymorphisms exist, that define different TPMT allozymes. Decreased TPMT enzyme activity can also be a consequence of lower protein synthesis, which depends on the promoter transcription activity. Promoter polymorphisms, such as variable number of tandem repeats (VNTR), can modulate the transcription. Administering thiopurine drugs in patients with certain genetic TPMT variants leads to severe hematologic toxicity. To avoid toxicity, therapy is being modified according to the TPMT genotype (pharmacogenetics). We investigated the polymorphisms in exons and regulatory elements (promoter) of the TPMT gene which affect TPMT enzyme activity in the Serbian population. We used PCR-based methodology and sequencing in the detection of genetic variants on TPMT gene. We showed that genetic variants in exons account for 7.5% of all TPMT variants with decreased enzyme activity. The therapy for patients with these pharmacogenetic markers was modified, which contributed to the efficiency of treatment. Functional assays in vitro showed that the TPMT promoter activity and, therefore, the quantity of TPMT protein synthesized, depended on the architecture of VNTRs (i.e. number and type) in the promoter. Promoter of the TPMT gene specifically responds to mercaptopurine treatment of K562 cells in a VNTR-dependent manner. Study of DNA-protein interactions revealed that Sp1 and Sp3 transcription factors interact with VNTRs. Our research pointed out that the VNTR promoter region of the TPMT gene could become a new pharmacogenetic marker, clinically significant for the individualization of thiopurine therapy

Imunoprofilaksa mastitisa krava izazvanog sa Staphylococcus aureus

Mastitis in cows represents one of the most actual problems in intensive dairy production. The prevention of pathogen penetration in the udder, its colonization and reproduction impose the constant need for regular milk check-ups, and preventive and therapeutic measures. Staphylococcus aureus causes subclinical and clinical mastitis, which when in the acute form can originate difficult and malignant udder infections with granulomatous and necrotic changes. Chronic forms of Staphylococcal mastitis most often pass as subclinical cases. An efficient commercial vaccine has not been introduced yet, though the application of autochthonous vaccines in the prevention of mastitis can give satisfactory results. In this study we have developed and applied an autochthonous vaccine prepared from S. aureus isolated from milk samples taken from an experimental farm, as well as from the referent capsular strain S. aureus. The vaccine was applied to experimental dairy cattle twice before calving in a dose of 5 mL. It consisted of inactivated bacterial S. aureus JR3 cells in a quantity of 1x1010 cfu/mL and 5 mg SM capsule of the strain S. aureus 2286. This vaccination of dairy cows has resulted in a significant decrease of subclinical and clinical manifested mastitis in the treated group, when compared to the vaccinated group of experimental animals.Mastitisi krava predstavljaju jedan od najaktuelnijih problema u intenzivnoj proizvodnji mleka. Sprečavanje prodora patogenog uzročnika u mlečnu žlezdu, njegovo naseljavanje i razmnožavanje, nameću stalnu potrebu za redovnom kontrolom mleka, kao i preduzimanje preventivnih i terapijskih mera. Staphylococcus aureus izaziva subkliničke i kliničke forme mastitisa, koje mogu u akutnoj formi da izazovu teške, maligne mastitise u vidu granulomatoznih i nekrotičnih promena. Hronične forme stafilokoknog mastitisa uglavnom često prolaze kao subklinički oblici obolenja mlečne žlezde Još uvek nije pronađena komercijalna efikasna vakcina protiv mastitisa izazvanog S. aureus-om, ali primena autohtonih vakcina u preveniranju mastitisa može dati zadovoljavajuće rezultate. Koristili smo autohtonu vakcinu koju smo pripremili od S. aureus-a izolovanog iz mleka uzetog sa ogledne farme i referentnog kapsularnog soja S. aureus. Vakcina je dvokratno aplikovana oglednim kravama dva meseca pred telenje u dozi od 5 ml, a sastojala se od inaktivisanih bakterijskih ćelija S. aureus JR3 u količini od 1x1010 cfu/ml i 5 mg SM kapsule soja S. aureus 2286. Ovaj način vakcinisanja krava protiv mastitisa izazvanog Staphylococcus aureus-om doveo je do smanjenja subkliničkih i kliničkih mastitisa u značajno manjem procentu u odnosu na kontrolnu grupu krava

Application of data mining in direct marketing in banking sector

The key of a successful business operation lies in a good communication with clients, and therefore companies are paying more attention to Customer Relationship Management. One strategy of the CRM is to analyze and understand the consumer’s behaviour and characteristics, and to reach the necessary answers based on the implementation of the direct marketing campaign. The aim of this study is to identify factors that would indicate the customers who are ready to submit long-term deposit to the bank. The obtained results will distinguish the group of clients who are satisfied with the bank's operations and are ready to participate in marketing campaigns. Having compared the methods used in the study, the method of classification has proved to be more reliable than others. This analysis obtains results through the use of data mining algorithm known as decision trees. The disadvantage of this method is inaccurate data supplied by the client

Genomics as a basis for precision medicine

Although medicine always aimed to be personalized, true implementation of personalized medicine in health care practice has started recently. Fascinating progress of molecular genetics has strongly contributed to this great achievement of modern medicine. Personalized medicine, also known as genome-based medicine and precision medicine, uses the knowledge of molecular basis of the disease in order to individualize treatment for each patient. Development of novel powerful high-throughput technologies has enabled better insight into “oms” landscape of many diseases, resulting in application of precision medicine approaches in their treatment. There are four cornerstones of modern precision medicine: “omics”-based diagnostics, pharmacogenomics, specific molecular targeted, gene and cellular therapy and predictive genetics. One of the most important successes of precision medicine is a discovery of novel diagnostic molecular markers. Furthermore, numerous newly discovered molecular markers have contributed to more precise classification of patients in distinct prognostic groups, leading to specific, more successful treatment protocols. Development of pharmacogenomics platforms and application of molecular–targeted therapy have led to the individualization of therapy, tailored to genetic profile of a disease in each patient. The development of gene therapies which can cure or prevent a disease by targeting disease-causing molecular defect has confirmed that the precision medicine has responded successfully to a great challenge. Additionally, cellular and tissue therapies have opened new possibilities for personalized treatment of many patients. Growing knowledge in predictive genetics leads to the preventive medicine, the most important goal of modern medicine. There is no doubt that we are getting closer to full implementation of precision medicine in every day clinical practice.Book of abstracts: International Conference of Biochemists and Molecular Biologists in Bosnia and Herzegovina - ABMBBIH May, 202

Molecular genetic markers as a basis for personalized medicine

Genetika i genomika su danas potpuno integrisane u medicinsku praksu. Personalizovana medicina, poznata i kao medicina zasnovana na genomu, koristi znanja o genetičkoj osnovi bolesti da bi se individualizovalo lečenje svakog pacijenta. Veliki broj genetičkih varijanti, molekularno-genetičkih markera, već se koristi u kliničkoj praksi za dijagnozu, prognozu i praćenje bolesti (monogenska nasledna oboljenja, fuzioni geni i rearanžmani u pedijatrijskim i adultnim leukemijama) i presimptomatsku procenu rizika od obolevanja (BRCA1/2 za kancer dojke). Osim toga, primena farmakogenomike u kliničkoj praksi značajno je doprinela individualizaciji terapije u skladu sa genotipom i profilom ekspresije gena pacijenta. Genetičko testiranje za nekoliko farmakogenomičkih markera (TPMT, UGT1A1, CYP2C9, VKORC1) obavezno je ili se preporučuje pre započinjanja terapije. Najvažniji doprinos medicine zasnovane na genomu je ciljana molekularna terapija, prilagođena genetskom profilu bolesti. Testiranje genetičkih varijanti u malignim oboljenjima (BCR-ABL, PML/RARa, RAS, BCL-2, KIT, PDGFR, EGF) doprinosi tačnijoj stratifikaciji različitih kancera i adekvatnom izboru terapije. Krajnji cilj medicinske nauke je da primeni gensku terapiju koja bi eliminisala uzrok bolesti ili prevenirala bolest, ciljajući genetički defekt koji leži u osnovi bolesti. Tehnologija koja prati gensku terapiju veoma se brzo razvija i već se uspešno primenjuje. Iako je medicina oduvek suštinski bila "personalizovana", prilagođena svakom pacijentu, personalizovana medicina danas koristi modernu tehnologiju i znanja iz oblasti molekularne genetike i genomike, omogućujući stepen personalizacije koji vodi ka značajnom napretku medicinske prakse.Nowadays, genetics and genomics are fully integrated into medical practice. Personalized medicine, also called genome-based medicine, uses the knowledge of the genetic basis of disease to individualize treatment for each patient. A number of genetic variants, molecular genetic markers, are already in use in medical practice for the diagnosis, prognosis and follow-up of diseases (monogenic hereditary disorders, fusion genes and rearrangements in pediatric and adult leukemia) and presymptomatic risk assessment (BRCA 1/2 for breast cancer). Additionally, the application of pharmacogenomics in clinical practice has significantly contributed to the individualization of therapy in accordance with the patient's genotype and gene expression profile. Genetic testing for several pharmacogenomic markers (TPMT, UGT1A1, CYP2C9, VKORC1) is mandatory or recommended prior to the initiation of therapy. The most important achievement of genome-based medicine is molecular-targeted therapy, tailored to the genetic profile of a disease. Testing for gene variants in cancer (BCR-ABL, PML/RARa, RAS, BCL-2) is part of the recommended evaluation for different cancers, in order to achieve better management of the disease. The ultimate goal of medical science is to develop gene therapy which will fight or prevent a disease by targeting the disease-causing genetic defect. Gene therapy technology is rapidly developing, and has already been used with success. Although medicine has always been essentially "personal" to each patient, personalized medicine today uses modern technology and knowledge in the field of molecular genetics and genomics, enabling a level of personalization which leads to significant improvement in health care

DEVELOPMENT OF EMOTIONAL COMPETENCE IN EXAM SITUATIONS WITH STUDENTS WHO ACHIEVE DIFFERENT ACADEMIC SUCCESS AND STUDY DIFFERENT STUDY PROGRAMS

The authors, in the theoretical part of the paper, define and explain the concept of emotional competence, as well as skills that should be developed and strengthened within the mentioned concept. Also, the importance of emotional competence in exam situations for the academic success of students has been presented, which is supported by the presentation of research related to this topic. The aim of the study was to examine whether students with different academic achievement and from different study programs showed different levels of emotional competence in exam situations. The research was conducted on a sample of 115 students from the Faculty of Philosophy, University of East Sarajevo. The paper presents the results of examining the differences in the development of emotional competence in exam situations in students with different academic achievement and the results of examining the differences in the development of emotional competence in students in exam situations depending on the study program they study. The findings show that there are no differences in the development of emotional competence in exam situations in students with different academic achievement, but there are differences when it comes to the development of emotional competence in exam situations depending on the study program that students’ study. This research is a contribution to a better understanding of students' emotional competencies in exam situations and may have practical implications in terms of understanding the importance of improving the emotional sphere in all study programs by expanding study objectives so that they include developing of students' emotional competencies. Article visualizations

Molecular characteristics, phenotypic diversity and genotype-estimated therapeutic responsiveness of Serbian patients with phenylketonuria

Fenilketonurija (PKU) jeste retko metaboličko oboljenje koje se nasleđuje autozomalno recesivno. Uzrok PKU su mutacije u genu koji kodira za enzim fenilalanin-hidroksilazu (PAH). U ovom revijskom radu su opisane najznačajnije metode za detekciju mutacija u PAH genu. Budući da slika o PKU prevazilazi "jednostavno" monogensko oboljenje, prikazani su i podaci o varijantama u nekodirajućim regionima PAH gena, kao i genima modifikatorima za PKU. Iako postoji značajna korelacija između PAH genotipa i PKU fenotipa, uočene su i neusaglašenosti. U ovom revijskom radu su izneti mogući uzroci za neusaglašenost između genotipa i fenotipa, kao što su: propust u detekciji više od dve mutacije prisutne u PAH genotipu pacijenta, neusaglašenost u fenotipskoj klasifikaciji pacijenata (koncentracija fenilalanina u serumu pre terapije i tolerancija fenilalanina), fenomen interalelske komplementacije (pozitivne i negativne). Novi terapijski pristup, terapija tetrahidrobiopterinom (BH4), predstavlja važnu inovaciju u lečenju pacijenata sa PKU. Međutim, u zemljama u kojima test opterećenja sa BH4 i terapija sa BH4 nisu dostupni, procena odgovora na terapiju koja je zasnovana na genotipu predstavlja koristan pristup. Ovakav pristup omogućava procenu potencijalne koristi od terapije sa BH4 što je bitna informacija, kako za pacijenta, tako i na nivou populacije. Predložen je i optimalni algoritam za molekularnu dijagnostiku, uspostavljen prema objavljenim učestalostima mutacija kod pacijenata sa PKU u Srbiji. U budućnosti, molekularno-genetički algoritam za PKU bi mogao da bude proširen tako da uključi i različite transkripcione regulatorne elemente u nekodirajućim regionima PAH gena, kao i gene modifikatore koji tek treba da budu otkriveni.Phenylketonuria (PKU) is a rare, inherited metabolic disease which is transmitted in an autosomal recessive pattern. PKU is caused by mutations in the gene encoding the phenylalanine hydroxylase (PAH) enzyme. This review cites the most prominent methods for the detection of mutations in the PAH gene. Since the image of PKU transcends "simple" monogenic disease, the known data on non-coding PAH gene variants and their role and PKU modifier genes have been further reviewed. It has been shown that there is a significant correlation between mutant PAH genotypes and PKU phenotypes. However, genotype-phenotype correlation inconsistencies have also been found. This review discusses the possible causes of phenotypic inconsistencies, such as oversight of more than two mutations present in the patient's PAH genotype, pitfalls of patient phenotypic classification (plasma phenylalanine concentration and phenylalanine tolerance), the inter-allelic complementation (positive and negative) phenomenon. A new therapeutic approach, tetrahydrobiopterin (BH4) supplementation therapy, is an important innovation in the course of PKU patients' treatment. However, in countries where the BH4-loading test and BH4-supplementation therapy are not available, a genotype-based estimation of responsiveness to the therapy is a valuable approach. It enables BH4-potential benefit estimation, which provides vital information both for the patient and for the population. An optimal molecular diagnostics algorithm, established according to the published mutation frequencies in Serbian PKU patients, has been suggested. In the future, the molecular-genetic algorithm for PKU could be expanded to include a variety of transcriptional regulatory elements located in noncoding PAH gene regions and yet to be discovered modifier genes