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Redefinisanje modela učešća građana u urbanističkom planiranju Srbije u skladu sa komunikativno kolaborativnom paradigmom

Redefining the model of citizen participation in urban planning in Serbia according to the communicative-collaborative paradigm

Planersku teoriju i praksu poslednjih petnaestak godina Karakteriše traganje za sistemom planiranja, koje je u procesu za štite i promovisanja zajedničkih vrednosti, sposobno za identifikaciju i osetljivo na aspiracije građana. Takav pristup planiranju nije nov, i po nekim karakteristikama se može uspostaviti relacija sa konceptima zastupničkog ili advokatskog planiranja formulisanim šezdesetih godina prošlog veka. Međutim, danas je, pored izuzetno visokog stepena raširenosti ideje o neophodnosti uključivanja širokog spektra aktera u proces planiranja, koncept izuzetno dobro i široko teorijski zasnovan, sa veoma rasprostranjenom i razvijenom proverom u praksi planiranja, tako da se može govoriti o novoj komunikativno-kolaborativnoj paradigmi planiranja. Termini koji se vezuju za različite varijetete planiranja sa naglašenim učešćem građana su: planiranje "odozdo-nagore" (eng. "bootom-up"), izvorno planiranje (eng. "grassroot"), participativno planiranje (eng. "participative planning"), demokratsko planiranje i planiranje sa zajednicom (eng. "community planning"). U radu se istražuje koncept planiranja u kojem se učešće građana, kao jedne od grupa aktera, smatra ne samo poželjnim, već neophodnim preduslovom dobrog planiranja i upravljanja razvojem. Traga se za odgovorima na pitanja: šta je to efikasno i efektivno učešće, ko su oni koji (bi trebalo da) učestvuju i na koji način to (bi trebalo da) čine, da li su informacione i komunikacione tehnologije (IKT) realna podrška na koju se možemo osloniti, i da li je i koliko je zaista moguće ostvariti savremen koncept učešća građana / planiranja, te time i potencijalno unaprediti urbanističko planiranje u Republici Srbiji. Istraživanje se odvija na dva nivoa: (1) Na nivou opštih konceptualnih opredeljenja, i njihove operacionalizacije do nivoa preporuka, baziranim na proučavanju relevantnih teorijskih radova i svetskih iskustava i primera u ovoj oblasti, a zatim, na osnovu toga, i na (2) Nivou urbanističkog planiranja u Srbiji, kao istraživačkom poligonu na kom se opšti nivo konceptualnih opredeljenja i njihove operacionalizacije proveravaju, a zatim i prilagođavaju formulisanjem modela planiranja adekvatnog istraživanom poligonu. Na osnovu istraživanja različitih teorijskih stanovišta, koncepata i praksi planiranja, prepoznate su mogućnosti i definisani načini i uslovi za realizaciju savremenog koncepta učešća građana u urbanističkom planiranju Srbije.

This work represents a contribution to the study of modern architecture and urbanism of the twentieth century in Belgrade. The paper investigates the architecture of public buildings on the principles of modern architecture as a significant and broad cultural phenomenon in the wider Belgrade and the Yugoslav territory in the period since 1918 untill 1941. During this period public buildings were built with the applied principles of modern architecture and urbanism. The work starts from the initial analysis of the modern movement of ideas between the two World Wars in Europe and the reflections that had the architects who were taking this model in the design and implementation of public buildings in Belgrade. Systematization and critical analysis of realized examples of public buildings to investigate the extent to which modern Belgrade was represented in the architectural production of the time, who were models from European experiences taken over directly and indirectly in the functional organization, volumetry architectural, design and materialization of objects. The work includes a chronological, critical analysis of the most important objects implemented in this period. The research starts from the premise that the preparation of the public buildings in Belgrade were present modern and avant‐garde ideas whose principles were established in the context of key historical, social and cultural phenomenon in Europe and Serbia. This attitude on the issue of research presupposes the establishment of correlation between the principles of modern architecture and implementation by local authors as well as theoretical reflection of changes that is generated in the development of modern Belgrade, which have promoted a new state and modern society...

Uloga urbanističkog planiranja u procesu regeneracije braunfild lokacija

The role of urban planning in the process of brownfield regeneration

Teza se bavi istraživanjem uloge urbanističkog planiranja u procesu regeneracije braunfild lokacija kao jednom od najsloženijih mehanizama održive upotrebe zemljišta. Specifičnost regeneracije braunfild lokacija se ogleda u posebnom načinu finansiranja tog procesa. Naime, dati proces zahteva sredstva javnog sektora, ali i privatnog kapitala, pri čemu privatni sektor često nije zainteresovan za ulaganje u braunfild lokacije. Ipak, kada se pomenuti akter odluči da učestvuje u procesu braunfild regeneracije, teško je uspostaviti kontrolu nad privatnim interesom, tj. balansiranje javnog i privatnog interesa postaje ključna veština u datom procesu. Pored uspostavljanja saradnje javnog i privatnog sektora, uspehu braunfild regeneracije doprinosi i integracija sektorskih politika iz različitih domena (saoobraćaja, životne sredine, lokalnog i regionalnog razvoja, itd.). S obzirom da se planiranje može razumeti kao jedan od oblika intervencije države na tržištu, fokus ovog istraživanja je na ispitivanju planskog pristupa procesu regeneracije braunfild lokacija. Osnovna istraživačka hipoteza se odnosila na prednost odvijanja procesa braunfild regeneracije u kolaborativnom modelu planiranja u odnosu na ostale planske modele (npr. racionalni). Pretpostavka je zasnovana na stavu da efikasan i efektivan proces donošenja odluka o braunfild regeneraciji zahteva usaglašenost interesa, kao i sinergiju znanja i veština čiji su nosioci zainteresovane strane koje učestvuju u datom procesu. Odavde je direktno proizašla sledeća hipoteza koja se odnosila na poziciju planerske ekspertize u procesu braunfild regeneracije. Dakle, pretpostavljeno je da je uspešna realizacija procesa braunfild regeneracije direktna posledica sinteze ekspertskih znanja (kao dominantne odlike racionalnog planskog modela), ali i veština pregovaranja, fasilitacije i medijacije, koje su imanentne kolaborativnom modelu planiranja. S obzirom da je planerska aktivnost neodvojiva od konkretnog konteksta u kojem je smeštena, važno je preispitivanje transformacije i prilagođavanja institucionalnog okvira kolaborativnom modelu planiranja, odakle je proizašla poslednja VI hipoteza. Stoga je pretpostavljeno da efektivan proces braunfild regeneracije zahteva formalni institucionalni okvir koji omogućava kolaboraciju i to: kolaboraciju sektora, disciplina i institucija na horizontalnom nivou, ali i na vertikalnim nivoima planiranja, kao i funkcionisanje javno-privatnih partnerstava. Pored toga, institucionalna transformacija podrazumeva podsticanje neformalnih planskih pristupa, odnosno inovativnih procedura, kojima se regulišu načini učešća čitavog spektra različitih stejkhodlera u planskom procesu. U teorijskom delu istraživanja je, kroz pregled planskih paradigmi, poseban akcenat stavljen na planiranje kao proces. Tačnije, posmatrao se način planiranja, odlučivanja i, posebno pozicija planerske ekspertize, s obzirom da se u odnosu na to očitava stepen i način kolaboracije učesnika u planskom procesu. Prikazu planskih paradigmi je prethodio kratak pregled modela upravljanja kao konteksta u kojem se dati planski modeli odvijaju. Među mnogobrojnim klasifikacijama načina upravljanja dat je prikaz tri modela koji su poslužili kao merilo za procenu upravljanja u realnom kontekstu, kao i za utvrđivanje pozicije različitih učesnika u ovom procesu. Naime, reč je o koordinativnom, kompetitivnom i komunikativnom (argumentativnom) modelu, sa glavnim akcentom na ulogama, odgovornostima i nadležnostima glavnih učesnika uključenih ili pogođenih upravljačkim procesom. S obzirom na porast internacionalizacije planiranja tokom XX veka, promena planske paradigme su posmatrane prvo u anglosaksonskom, kao razvijenom kontekstu, da bi zatim bile prikazane specifične karakteristike paradigmatske promene u okviru postsocijalističkih država. U drugom delu teorijskog okvira istraživanja pažnja je posvećena kolaborativnom planskom modelu i temama koje su od značaja za odvijanje procesa braunfild regeneracije u okviru tog modela - kolaboraciji stejkholdera (zainteresovanih aktera), usaglašenosti interesa, rešavanju konflikata, postizanju konsenzusa, veštinama i znanjima planera, načinima implementacije, itd. Naime, radi razrade osnovnih hipotetičkih aspekata čitavog istraživanja (efektivnih načina kolaboracije, adekvatne planerske ekspertize, kao i optimalnih institucionalnih aranžmana) dat je prikaz osnovnih koncepata izabrane tri teorije koje posebno odgovaraju zahtevima koje postavlja problem VII braunfild regeneracije. Dakle, za temu kolaboracije posebno se izdvaja teorija aktera-mreže, koja govori upravo o formiranju mreže, kao osnovnom obliku organizacije učesnika u složenim procesima; sa aspekta planerske ekspertize, tj. znanja i veština planera, od značaja je teorija pregovaranja, kojom se naglašava značaj eksperata u pregovaranju kao sastavnom delu kolaborativnog procesa, dok o aspektu institucionalnog okvira razvojnih procesa govori institucionalna teorija. Ova teorijska razmatranja čine osnovu za definisanje generalnih istraživačkih pitanja, koja su potom razmatrana kao principi konceptualnog modela kolaboracije stejkholdera u procesu braunfild regeneracije. Poseban doprinos ovog modela se ogleda u tome što on zapravo predstavlja sredstvo evaluacije različitih pristupa braunfild regeneraciji u okviru izabranih primera koji su prikazani u praktičnom delu istraživanja. Metodologija rada je prilagođena generalnim istraživačkim pitanjima, hipotezama i ciljevima istraživanja. Preciznije, praktični cilj istraživanja je utvrđivanje i prikaz načina regeneracije braunfild lokacija uz istovremenu proveru stepena u kojem je primenjeni pristup zaista kolaborativan. Prostorni obuhvat istraživanja se odnosi na zemlje koje teritorijalno pripadaju dunavskom basenu - Švajcarsku, Austriju, Češku, Mađarsku i, na kraju, Srbiju. Kako se ovim istraživanjem ne teži prikazu samo jednog primera regeneracije braunfilda, već izvođenju generalnih zaključaka analizom višestrukih empirijskih podataka, nameće se studija višestrukog slučaja kao odgovarajuća metoda. Sa aspekta ciljeva ovog istraživanja, kao najvažnije metode sadržane u okviru metode studije slučaja su: 1. analiza sadržaja primarnih i sekundarnih izvora koji se tiču institucionalnog konteksta (institucionalne strukture i dokumenata) procesa braunfild regeneracije, čime je zapravo proveravano da li konkretan institucionalni kontekst omogućava i podstiče kolaborativni planski pristup, i 2. ispitivanje izabranih stejkholdera, što je korišćeno za utvrđivanje načina kolaboracije među dominantnim stejkholderima, pozicije planerske ekspertize i prisustva neformalnih pristupa u konkretnom procesu braunfild regeneracije. Dakle, suština metodološkog pristupa se odnosi na ispitivanje kolaborativnih procedura prilikom braunfild regeneracije. Istraživanjem je potvrđena teza da proces regeneracije braunfild lokacija zahteva odvijanje u okviru kolaborativnog planskog modela. Takođe se potvrđuje VIII teza o ekspertnosti planera kao sintezi ekspertskih znanja, sa jedne strane, ali i veština kolaboracije i posredovanja u procesu braunfild regeneracije, sa druge. Na kraju, kolaboracija različitih sektora koje si u nadležnosti javne uprave na horizontalnom nivou, ali i među različitim vertikalnim nivoima planiranja, kao i institucionalizacija neformalnih planskih pristupa doprinosi zadovoljavajućem planskom ishodu procesa braunfild regeneracije. Doprinos istraživanja je dvojak. Prva grupa rezultata se odnosi na istraživanje razvoja teorijsko-metodološkog pristupa za formulisanje principa braunfild regeneracije. Pored toga, konceptualni model kolaboracije stejkholdera u procesu braunfild regeneracije predstavlja mehanizam evaluacije različitih pristupa datom procesu. Druga grupa rezultata je praktičnog tipa i odnosi se na evropska iskustva, kao i iskustva Srbije u braunfild regeneraciji. U kontekstu inostranih primera braunfild regeneracije doprinos istraživanja se, kao prvo, odnosi na prikaz načina planiranja i odlučivanja u konkretnom institucionalnom kontekstu - razvijenim, kao i postsocijalističkim zemljama. Pored toga, svaka od analiziranih studija slučaja nudi efikasne modele (kolaboracije stejkholdera, planerske ekspertize i institucionalne organizacije) za pristup problemu braunfild regeneracije. Poseban doprinos istraživanja je sagledavanje veze između institucionalnog i legislativnog okvira i konkretnog slučaja braunfild regeneracije. Ključni doprinos istraživanja je pregled stanja braunfild regeneracije na području Srbije i, posebno, procena uticaja institucionalnog i legislativnog okvira na planiranje procesa braunfild regeneracije, kao i pružanje smernica za dati proces. Time se, zapravo, teži redefinisanju karaktera urbanističkog planiranja u domenu regeneracije braunfild lokacija u Srbiji.

The thesis aims at researching the role of urban planning in the process of brownfield regeneration, as one of the most complex sustainable land-use mechanisms. The peculiarity of brownfield regeneration is best seen in the specific way of financing of that process. Namely, the mentioned process requires the funding of public sector and also private capital, but not always does private sector show interest in investing in brownfield land. Nevertheless, whenever this actor decides to take part in the process of brownfield regeneration, private interest becomes difficult to control i.e. the crucial skill in this process is seen as sole balancing between public and private interests. In addition to establishing the cooperation between public and private sector, what also contributes to success of brownfield regeneration is the integration of sector policies focusing on different domains (traffic, environment, local and regional development, etc.). Since planning can be understood as a form of state intervention in the market, this research focuses on the assessment of planning reproach to the process of brownfield regeneration. The basic research hypothesis referred to the advantage of this process being realized through the means of collaborative planning model, comparing to other models (e.g. rational planning model). This assumption was based on a view that an efficient and effective decision-making process about brownfield regeneration needed harmonised interests, as well as synergy of knowledge and skills, and is carried out by interested parties which are involved in a given process. This leads to the next hypothesis which referred to the position of planning expertise in the process of brownfield regeneration. Hence, it was assumed that a successful realisation of brownfield regeneration process was a direct consequence of the synthesis of experts' knowledge (as the main feature of rational planning model), but also the skills of negotiation, facilitation and meditation ingrained in collaborative planning model. Considering the fact that planners' activity is inseparable from the specific context in which it takes place, it is important to review transformation and adaptation of the institutional framework to collaborative planning model, which leads to the next hypothesis. Therefore, it was presumed that an effective process of brownfield regeneration required a formal institutional framework which enables collaboration, as X follows: collaboration of sectors, disciplines and institutions at the horizontal level, but also at the vertical planning levels, as well as the functioning of public-private partnerships. Besides, institutional transformation means incentive to informal planning approach and innovative procedures to regulate the participation of a range of different stakeholders in the planning process. Planning as a process was especially emphasised through planning paradigm review in the theoretical part of the research. More precisely, what was being observed was the way of planning, decision-making and especially planning expertise position, since the degree and manner of participants' collaboration in the planning process is to be determined with respect to it. Before planning paradigm review, a short review of the management model, as the context in which given planning models take place, was given. Among numerous classifications of management types, there were three models given. They could serve as a measure for management estimation in the real context, but could also serve for defining positions of different participants in this process. Namely, these are coordinative, competitive and communicative (argumentative) model, mainly stressing the roles, responsibilities and accountabilities of the main participants included and influenced by management. With regard to the growth in internationalisation of planning throughout the 20th century, planning paradigm shift was firstly analysed in the Anglo-Saxon i.e. developed context, followed by the review of the specific characteristics of paradigm change in post-socialist countries. In the second part of theoretical research framework the attention was being given to collaborative planning model and subjects which were relevant to the process of brownfield regeneration within this model – collaboration of stakeholders, harmonisation of interests, conflict solving, reaching a consensus, planners' skills and knowledge, different ways of implementation, etc. For the reason of elaboration of basic hypothetical aspects of the overall research (the effective ways of collaboration, the adequate planning expertise, as well as optimal institutional arrangements), the basic concepts of the three chosen theories which meet demands put forward by the brownfield regeneration problem were reviewed. Therefore, for the subject of collaboration especially important is the actor-network theory, dealing with network- forming as a basic form of participant organisation in a complex processes. From the perspective of planning expertise, i.e. planners' knowledge and skills, the theory of negotiation is important, since it stresses the relevance of experts in negotiation as a part XI of collaborative process, whereas institutional theory deals with the aspect of institutional framework of developing processes. These theoretical arguments represent the basis for defining general research questions, which were afterwards analysed as principles of conceptual model of stakeholders' collaboration in the process of brownfield regeneration. Special contribution of this model lies in the fact that it actually represents means of evaluation of different approaches to brownfield regeneration, within the chosen examples shown in the practical part of the research. The methodology of the research is adjusted to general research questions, hypothesis and research goals. More precisely, the practical goal of the research is to establish and represent ways of brownfield regeneration, at the same time reviewing to which degree is the applied principle actually collaborative. The spatial coverage of the research involved the countries which territorially belong to the Danube basin – Switzerland, Austria, the Czech Republic, Hungary, and finally, Serbia. Since this research does not aim at representing only one example of brownfield regeneration, but at reaching general conclusions through analysis of the multiple empirical data, a multiple case study was being imposed as the adequate method. Having in mind the goals of this research, the most important methods used within the case study were: 1. analysis of the content of primary and secondary sources which are related to institutional context of brownfield regeneration process (institutional structure and documents) by which it was actually being checked whether certain institutional context enabled and encouraged collaborative planning approach; 2. assessment of chosen stakeholders, used for establishing the ways of collaboration among dominant stakeholders, positions of planning expertise and presence of informal approach in the concrete process of brownfield regeneration. Hence, the essence of methodological approach was related to the revision of collaborative procedures during brownfield regeneration. Research confirmed the thesis that brownfield regeneration asked for realization within collaborative planning model. The thesis on planers' expertness as the synthesis of experts' knowledge, on the one hand, and the collaboration and mediation skills throughout the process of brownfield regeneration, on the other hand, is confirmed. Finally, the collaboration of different sectors under jurisdiction of public administration at the horizontal level, but also between different vertical levels of planning, as well as the institutionalization of informal planning approach contributes to the satisfactory outcome of the process of brownfield regeneration. XII The contribution of the research is twofold. The first group of results is related to the development of theoretical and methodological approach for formulation of principles in brownfield regeneration. Apart from that, conceptual model of stakeholders' collaboration in the process of brownfield regeneration represents evaluation mechanism of different approaches to a given process. The second group of results is practical, and refers to European experiences, but also to experiences of brownfield regeneration in Serbia. In the context of foreign brownfield regeneration examples, the contribution of the research primarily refers to the representation of planning and decision making in the specific institutional context – in developed, as well as post- socialist countries. Additionally, each of the analysed case studies offers the efficient models (stakeholders' collaboration, planning expertise and institutional organisation) for brownfield regeneration approach. A significant contribution made by the research is the review of the current state of brownfield regeneration in Serbia and, especially, the estimation of the influence of institutional and legislative framework on the planning of brownfield regeneration, as well as providing guidelines for the given process. The goal here is to actually redefine the character of urban planning in the domain of brownfield regeneration in Serbia.

Morfologija gradskog bloka - analiza mogućnosti primene u projektovanju i planiranju

Morphology of a city block - possibilities for applying in designing and planning

U ovom radu se ukazuje na nužnost uspostavljanja dijaloga između sveobuhvatnih morfoloških istraživanja i prakse projektovanja i planiranja, gde se mogućnosti povezivanja procesa teoretske elaboracije i praktične primene tumače kroz istraživački koncept i fenomen gradskog bloka koji sa jedne strane predstavlja karakterističnu morfološku celinu u sklopu grada, a sa druge strane urbani nivo na kome se direktno ispoljavaju efekti i međuuslovljenosti procesa projektovanja i planiranja. Morfogenetskom analizom gradskog bloka mogu se detektovati karakter i uzroci prirodnih ili planski usmerenih procesa razvoja grada i obrnuto: na osnovu analize postojećeg stanja i savremenih razvojnih procesa moguće je pretpostaviti odgovarajuće oblike fizičke strukture i interpretirati ih na nivou bloka. U tom smislu blok može predstavljati istraživački koncept u procesima arhitektonsko‐ urbanističkog projektovanja i urbanističkog planiranja, kako u fazi analize postojećeg stanja, tako i u završnoj fazi davanja planskih smernica u obliku preporuka za pravila uređenja i građenja ili definisanja modela. Formiranje konceptualnih modela gradskog bloka predstavlja vizuelizaciju ishoda morfoloških i kontekstualnih istraživanja i smernicu ka ekstrapolaciji teoretskih istraživanja u projektantsku i planersku praksu i obrnuto: praksa predstavlja bazu i materijal akumuliranog znanja, veština, inovacija arhitekata, planera, učešća svih aktera koja se istraživanjem i vrednovanjem ugrađuje u teoriju i predstavlja polazište za razvoj novih koncepcija i modaliteta u operacionalizaciji.

This dissertation indicates the need to establish a dialogue between comprehensive morphological investigations and practice in planning and design where the possibilities for linking the process of theoretical elaboration and practical implementation are interpreted through a research concept and phenomenon of city block which represents a typical morphological entity, on the one hand, and an urban level at which effects and interdependence of planning and design process manifest themselves, on the other hand. Through a morphogenetic analysis of an city block, it is possible to identify a character and causes of natural processes or directed planning processes in urban development, and vice versa: based on an analysis of the existing state and contemporary processes of urban development, it is possible to presume appropriate shape of physical structure and interpret it at the level of block. In this context, the block can represent a research concept in architectural and urban planning processes both in the stage of the analysis of the existing state and in the final stage of setting the guidelines in the form of recommendations for planning and building rules or for defining a model. The creation of conceptual models of city block implies a visualization of outcomes of morphological and contextual investigations and a guideline towards extrapolation of theoretical research into planning and design practice and vice versa: practice is a basis and material of accumulated knowledge, skills, innovations of architects, planners, involvement of all actors, which is incorporated into the theory through research and evaluation and represents a starting point for the development of new concepts and modalities in operationalization.

Anamorfoza i polni dimorfizam balkanskog endemita Apfelbeckia insculpta (L. Koch, 1867) (Diplopoda, Callipodida, Schizopetalidae)

Anamorphosis and sexual dimorphism in the Balkan endemic species Apfelbeckia insculpta (L. Koch, 1867) (Diplopoda, Callipodida, Schizopetalidae)

Ova teza je fokusirana na razrešavanje fenomena koji su vezani za postembrionalno razviće i polni dimorfizam kod balkanske endemične stonoge Apfelbeckia insculpta (L. Koch, 1867), jedne od najvećih evropskih stonoga, čija je distribucija ograničena na teritoriju Srbije, Crne Gore, Bosne i Hercegovine, Hrvatske i Albanije. Analizirani takson pripada grupi troglofilnih vrsta; najčešće naseljava pećine i druga endogejska staništa, ali ponekad može biti prisutna i u epigejskim ekosistemima. Prvi deo teze se odnosi na analizu postembrionalnog razvića kod ispitivane vrste. Kao i kod drugih Myriapoda i Arthropoda, postembrionalni period života kod A. insculpta podrazumeva postojanje različitih stadijuma čije je trajanje ograničeno procesom presvlačenja. Na osnovu uzoraka prikupljenih iz pedeset populacija sa teritorija Srbije, Crne Gore, Bosne i Hercegovine, i Hrvatske, ustanovljeno je da A. insculpta tokom postembrionalnog razvića prolazi kroz deset stadijuma. Svako presvlačenje je praćeno povećanjem broja pleurotergita sa i bez nogu za hodanje, broja parova nogu za hodanje i broja redova ocela. Analizom navedenih struktura kod svake jednike ponaosob, bilo je moguće precizno identifikovati anamorfni stadijum kome ta jedinka pripada. Način postembrionalnog razvića kod A. insculpta je teloanamorfoza, što podrazumeva da se sa finalnim presvlačenjem (devetim kod analizirane vrste) uspostavlja adultni stadijum, jedinke postaju polno zrele i nakon toga ne postoje dodatna presvlačenja. Mužjaci i ženke se mogu razlikovati počevši od stadijuma VIII. Drugi deo teze posvećen je analizama fenomena koji su vezani za polni dimorfizam ispitivanog taksona. Analiza polnog dimorfizma je obuhvatila četiri populacije iz Srbije i jednu iz Crne Gore. Razlike u veličini i proporcijama tela su ispitivane univarijantnim i multivarijantnim statističkim metodama. Pokazano je da su ženke krupniji pol (u okviru svih pet populacija statistički značajno veće vrednosti dužine i mase tela su bile prisutne kod ženki). Takođe, dimenzije glave (dužina i širina) i trupa (dužina, širina i visina) su bile veće kod ženki iz svih uzorkovanih populacija. Dužine antena i nogu za hodanje sa sredine trupa su pokazale suprotan smer ekspresije polnog dimorfizma, tj. veće vrednosti su registrovane kod mužjaka iz svih populacija. Razlike u obrascu ekspresije među populacijama su utvrđene za dimenzije gnatohilarijuma (dužinu i širinu), kao i dužine prvog, drugog i četvrtog para nogu, i podrazumevale su da su veće vrednosti dužine i širine gnatohilarijuma prisutne kod ženki iz dve populacije, dok su dužine navedena tri para nogu bile statistički značajno veće kod ženki u okviru samo jedne populacije. U ostalim slučajevima, interseksualne razlike u merama gnatohilarijuma i anteriornih nogu nisu bile statistički značajne. Generalno, kod ženki su prisutne veće vrednosti karaktera koji opisuju veličinu celog tela, kao i onih koji opisuju veličinu svakog telesnog regiona (glave i trupa), dok su kod mužjaka veće vrednosti registrovane za karaktere koji se mogu povezati sa pokretljivošću organizma, pronalaženjem i prepoznavanjem partnera za kopulaciju, kao i ponašanjem tokom samog čina parenja. Takođe, utvrđeno je da stepen polnog dimorfizma varira među analiziranim populacijama. Pretpostavljamo da su selekcija vezana za fekunditet i seksualna selekcija glavni mehanizmi koji utiču na veličinu i proporcije tela kod mužjaka i ženki A. insculpta. Pored analize polnog dimorfizma kod adultnih jedinki, praćene su i morfološke razlike između mužjaka i ženki tokom ontogenije. Ova analiza je obuhvatala tri postembrionalna stadijuma tokom kojih je moguća distinkcija polova (stadijumi VIII, IX i adulti). Ispitivanja promena u veličini i proporcijama tela tokom postembriogeneze su podrazumevala univarijantne, bivarijantne i multivarijantne statističke analize. Rezultati ovih analiza su pokazali da se u osnovi polnog dimorfizma nalaze razlike u stopama rasta. Više stope rasta dužine i mase tela, dužine i širine glave, kao i dužine, širine i visine trupa su prisutne kod ženki. Suprotno ovim karakterima, više vrednosti stopa rasta antena i nogu za hodanje su prisutne kod mužjaka. Pored razlika u stopama rasta, utvrđeno je da polni dimorfizam u dužini glave nastaje kao posledica različitih alometrijskih obrazaca kod mužjaka i ženki (kod ženki je prisutna izometrija, a kod mužjaka negativna alometrija). Za razliku od ostalih karaktera, kod dužine nogu je utvrđeno da dolazi do promene u smeru, a i stepenu polnog dimorfizma tokom ontogenije. Na stadijumu VIII su veće vrednosti dužine nogu prisutne kod jedinki ženskog pola, dok su na stadijumu IX, kao i na adultnom stadijumu veće vrednosti dužine nogu za hodanje prisutne kod mužjaka. Ovakav rezultat verovatno nastaje usled različitih selekcionih pritisaka u okviru svakog stadijuma.

This dissertation is focused on resolving phenomena related to post-embryonic development and sexual dimorphism in the Balkan endemic millipede Apfelbeckia insculpta (L. Koch, 1867) which is one of the largest of European millipedes and is distributed in Serbia, Montenegro, Bosnia and Herzegovina, Croatia, and Albania. The analyzed taxon is classified as a troglophilic species, i.e. it usually inhabits caves and other underground habitats, but sometimes can be found in epigeic habitats. Analysis of post-embryonic development is a focal point of the first part of this doctoral dissertation. As in other myriapods and arthropods, post-embryogenesis of A. insculpta is composed of different stadia which are separated by molts. Based on the data gathered from fifty populations from Serbia, Montenegro, Bosnia and Herzegovina, and Croatia, we conclude that during post-embryonic period of the life cycle A. insculpta undergo ten different stadia. Each molt is accompanied by the addition of podous and apodous pleurotergites, leg pairs and rows od ocelli. Thus, the analysis of these structures provided system for precise identification of anamorphic stadium of each individual. The mode of post-embryonic development in A. insculpta is teloanamorphosis, i.e. with the ninth and the last molt individuals become adults, reach sexual maturity and no additional molts occur after this. Males and females can be distinguished from stadium VIII onward. The second part of this dissertation is focussed on sexual dimorphism in A. insculpta. Data for analysis of sexual dimorphism are gathered from four populations in Serbia and one in Montenegro. Univariate and multivariate statistical techniques are used for analyses of intersexual differences in both body size and body proportions. Females are the larger sex (in all analyzed populations, females expressed larger values for body length and body mass). Also, measures of head length, head width, trunk length, trunk width, and trunk height were significantlly higher in females in all analyzed populations. Comparisons of length of the antennae and length of the walking legs showed that the opposite pattern of sexual dimorphism is present in A. insculpta, i.e. dimorphism was male-biased for these characters in all populations. Sexual dimoprhism in gnathochilarium measurements (length and width), as well as in lengths of the first, second and fourth leg pairs showed inter- populational differences and both gnathochilarium measures had significantly higher values in females originating from two populations, while the mentioned three pairs of anterior legs were larger in females from only one population. Analyses showed that differenes between males and females for gnathochilarium and anterior legs measurements were nonsignificant in remaining populations. Generally, females had significantly greater body size than males, as well as greater both body regions (head and trunk), while males expressed greater values in traits that can be associated with mobility, mate search, and copulation behavior. Also, we found significant variations in the degree of sexual dimorphism among analyzed populations. The influences of fecundity and sexual selection on females and males are regarded as the most probable drivers that affect variation in adult morphology in our study species. In addition to the analysis of sexual dimorphism within adult stadium, we were focussed on ontogenetic perspective of sexual dimorphism in A. insculpta. Animals used in this analysis belonged to three post-embryonic stadia (VIII, IX, and adults) where sexes could be distinguished. Univariate, bivariate, and multivariate statistical procedures were used for quantification of interexual differences in body size and body proportions during ontogeny. Our results indicate that differences between sexes arise via different growth rates. Females exhibit higher growth rates in body length, body mass, head length, head width, trunk length, trunk width, and trunk height. On the other hand, higher growth rates in males are present in lengths of the antennae and the walking legs. Apart from growth rates, our results showed that sexual dimorphism in head length arise beacuse different allometric trends are present in males and females (negative allometry in males and isometry in females). In contrast to all other characters, it is shown that ontogenetic shift in the direction of sexual dimorphism occurs during the development of walking legs. Sexual dimorphism was female-biased for this character at stadium VIII, while the opposite patern of sexual dimorphism in length of the walking legs is present in stadium IX and in adult millipedes. We suppose that variation in selection pressures during ontogeny are the underlynig factors that drive such pattern of sexual dimorphism.

Imunski mehanizmi indukcije i ekspresije reakcije kontaktne preosetljivosti kod pacova

Immune mechanisms of induction and expression of contact hypersensitivity reaction in rats

Alergijski kontaktni dermatitis (AKD) je česta zapaljenska bolest kože kod ljudi, koja pogađa 15-20% svetske populacije. Velika zastupljenost hemijskih agenasa (haptena) koji izazivaju AKD, uz odsustvo adekvatne konvencionalne imunosupresivne terapije, čine ovu bolest velikim izazovom za javno zdravlje, i savremenu imunologiju. Razvoj životinjskog modela AKD, reakcije kontaktne preosetljivosti, omogućio je napredak u razumevanju njene patogeneze. Iako je reakcija kontaktne preosetljivosti (KP) dosta ispitivana, pretežno na mišjem modelu, mehanizmi u osnovi razvoja bolesti nisu dovoljno poznati. Takođe, mali je broj podataka o reakciji KP kod drugih eksperimentalnih životinja, osim miševa, koji bi mogli da unaprede znanje o imunobiologiji kontaktnih alergija. U svetlu toga, u ovom radu su ispitane karakteristike reakcije KP kod pacova, korišćenjem dva soja pacova, Dark Agouti (DA) i Albino Oxford (AO), za koje je pokazano da se razlikuju u reaktivnosti na pojedine zapaljenske stimuluse. Indukcija (senzibilizacija) i izazivanje (elicitacija) reakcije KP vršene su sa haptenom dinitrohlorobenzenom (DNCB), koji je primenjen u dve različite koncentracije (režim senzibilizacije/elicitacije 0.4%/0.13% i 4%/1.3% DNCB). Mehanizmi KP kod pacova ispitani su u fazi senzibilizacije i elicitacije reakcije, praćenjem dinamike promena aktivnosti ćelija drenirajućih limfnih čvorova (dLČ) u fazi senzibilizacije, i ispitivanjem produkcije inflamatornih/ anti-inflamatornih medijatora u kondicioniranom medijumu organokulture uha, i promena aktivnosti ćelija dLČ u fazi elicitacije reakcije (24 časa nakon elicitacije). Rezultati su pokazali povećanje in vivo (otok uha) i ex vivo (patohistološka analiza isečaka kože uha) parametara reakcije KP kod oba soja pacova nakon tretmana sa višom dozom haptena, ali je reakcija bila intenzivnija kod DA u odnosu AO soj. Nakon senzibilizacije dolazi do: povećanja broja ćelija u dLČ kod oba soja (izraženije kod DA jedinki), promene zastupljenosti CD4+ i CD8+ ćelija i njihovog odnosa (povećanja odnosa CD4+/CD8+ ćelija kod DA soja, zbog povećanja zastupljenosti CD4+ ćelija a pada zastupljenosti CD8+ ćelija, i smanjenja odnosa CD4+/CD8+ kod AO soja, zbog pada zastupljenosti CD4+ ćelija a povećanja CD8+ ćelija), povećanja ex vivo proliferacije izolovanih ćelija i povećanja spontane i specifične (nakon in vitro stimulacije ćelija sa haptenom) produkcije inflamatornih citokina IFN-γ i IL-17, i Th2/anti-inflamatornog citokina IL-10 (izraženije kod DA soja). U ovoj fazi reakcije kod oba soja nije zapažena promena u produkciji i ekspresiji Th2/anti-inflamatornog citokina IL-4. U fazi ekspresije (elicitacije) reakcije pokazano je da je povećanje otoka uha praćeno povećanjem produkcije TNF-α, NO, MPO, IFN-γ i IL-17 u kondicioniranom medijumu organokulture eksplanata kože uha, kao i da su razlike u ekspresiji reakcije između dva soja praćene razlikama u intenzitetu promena (veća koncentracija NO, MPO, IFN-γ, IL-17 kod DA soja) i tipu imunskog odgovora (povećana produkcija IL-4 samo kod AO soja). U ovoj fazi reakcije pokazano je povećanje aktivnosti ćelija dLČ (spontane i specifične proliferacije, uz povećanje ekspresije inducibilnog α lanca receptora za faktor rasta IL-2, kao i produkcije efektorskih citokina IFN-γ i IL-17, i ekspresije gena za p35, p40 i T-bet, koji su pokazatelji diferencijacije Th1/tip1 ćelija) kod DA soja nakon tretmana sa DNCB-om, u odnosu na kontrole. Odsustvo promena u aktivnosti efektorskih mehanizama u dLČ kod jedinki AO soja je praćeno povećanom produkcijom IL-4, i povećanim brojem CD4+CD25+Foxp3+ ćelija, koje fenotipski odgovaraju regulatornim T ćelijama (Treg). Reakcija KP na DNCB kod pacova (slično mišjem modelu) karakteriše se aktivacijom Th1/tip1 i Th17/tip imunskog odgovora, a razlike u intenzitetu odgovora (u fazi senzibilizacije i elicitacije) ili tipu odgovora (u fazi elicitacije), mogu biti u osnovi zabeleženih međusojnih razlika u intenzitetu ekspresije reakcije. Komparativno ispitivanje kontaktne preosetljivosti kod reaktivnijeg DA soja i manje reaktivnog AO soja, može doprineti saznanjima o imunskim mehanizmima koji su u osnovi razvoja i ekspresije, kao i regulacije ove reakcije. Ovakva saznanja mogu dati dobru osnovu za razvoj novih terapeutskih modaliteta u lečenju alergijskog kontaktnog dermatitisa, ali i osnovu za ispitivanje imunotoksičnog potencijala hemijskih agenasa, posebno onih koji dospevaju u organizam preko kože.

Allergic contact dermatitis (ACD) is a common skin inflammatory disease in humans, affecting 15-20% of the general population. Large number of chemical agents that induce ACD (called haptens), together with lack of adequate conventional immunosuppressive therapy, make this disease a major challenge for public health and modern immunology. Pathophysiology of ACD is studied on animal models referred as contact hypersensitivity. Although the reaction of contact hypersensitivity (CHS) has been much studied, mostly in mice, the knowledge of its mechanisms is still far from complete. In addition, there is paucity of data about CHS in other experimental animals, beside mice, which might increase our understanding of the immunobiology of contact allergy. In light of this, in the present study the characteristics of CHS were analyzed in rats, using two strains, Dark Agouti (DA) i Albino Oxford (AO), which have been shown to differ in reactivity to certain inflammatory stimuli. CHS to hapten dinitrochlorobenzene (DNCB) was induced by applying two different hapten concentrations in sensitization and elicitation phases (sensitization/elicitation regime 0.4%/0.13% and 4%/1.3% DNCB). Mechanisms of CHS were studied in induction (sensitization) and expression (elicitation) phases, by evaluating dynamics in activity changes of draining lymph node (dLN) cells in sensitization phase and by evaluating production of inflammatory/anti-inflammatory mediators in conditioned medium of ear skin organ culture and activity changes of dLN cells in elicitation phase (24 hours after elicitation). The results showed increased in vivo (ear thickness) and ex vivo (patohistology analysis of ear skin) parameters of CHS reaction, in both rat strains following the treatment with high DNCB dose, but the reaction was more intensive in DA compared to AO rats. In sensitization phase it was showed: increased dLN cell number in both strains (higher in DA rats), changes in ratio of CD4+ and CD8+ subsets (increased ratio of CD4+/CD8+ cells in DA rats, because of increased relative number of CD4+ and decreased relative number of CD8+ cells and decreased CD4+/CD8+ ratio in AO rats, because of decreased relative number of CD4+ cells and increased CD8+ cells relative number), increased ex vivo proliferation of dLN cells and increased spontaneous and specific (induced by in vitro cell stimulation with hapten) production of inflammatory cytokines IFN-γ and IL-17 and Th2/anti-inflammatory cytokine IL-10 (more pronounced in DA rats). Similarly to DA rats, no changes in production and expression of Th2/anti-inflammatory cytokine IL-4 were noted in AO rats in sensitization phase. Increased ear thickness in elicitation phase was accompanied by increased production of TNF-α, NO, MPO, IFN-γ and IL-17 in conditioned medium of ear skin organ culture. Strain-dependent differences in CHS expression were related to differences in intensity (higher NO, MPO, IFN-γ, IL-17 in DA strain) and type of immune response (increased production of IL-4 only in AO rats). In addition, in this phase of the reaction it was showed the increase in dLN activity in DNCB-treated DA rats (increased spontaneous and specific proliferation, concomitantly with increased expression of growth factor IL-2 receptor α chain, as well as production of effector cytokines IFN-γ and IL-17, and gene expression of p35, p40 and T-bet, which are indicators of differentiation of Th1/type1 cells), compared to controls. The absence of changes in the activity of effector mechanisms in dLN of AO strain is accompanied by increased production of IL-4 and increased numbers of CD4+CD25+Foxp3+ cells, that phenotypically correspond to regulatory T cells (Treg). CHS to DNCB in rats (similarly to mouse model) is characterized by activation of Th1/type1 and Th17/type17 immune response, and differences in response intensity (in induction and expression phase) and response type (in expression phase) could be underlying mechanism of strain-dependent difference. Comparative analysis of CHS in more reactive (DA) and less reactive (AO) strains, can contribute to the further knowledge of the immunological mechanisms underlying the development and expression, as well as regulation of this reaction. Such findings might be useful in exploring the potential of therapeutic modalities to reduce skin damage caused by contact allergy, as well as in the elucidation of immunomodulatory potential of different chemicals, particularly those that gain access to the host system via skin.

Genotyping, antibiotic resistance and biofilm formation of Stenotrophomonas maltophilia clinical isolates from Serbia

Генотипизација, резистенција на антибиотике и формирање биофилма клиничких изолата Stenotrophomonas maltophilia из Србије

Genotipizacija, rezistencija na antibiotike i formiranje biofilma kliničkih izolata Stenotrophomonas maltophilia iz Srbije Stenotrophomonas maltophilia je sredinska bakterija i oportunistički patogen koji se najčešće povezuje sa nozokomijalnim infekcijama, a od skoro je svrstan u grupu globalnih patogena koji su višestruko rezistentni na antibiotike. Cilj ove doktorske disertacije jeste gneotipizacija i fenotipska karakterizacija kolekcije S. maltophilia koja je sakupljena tokom svakodnevne zdravstvene nege pacijenata u Institut za zdravstvenu zaštitu majke i deteta „dr Vukan Čupić“. To je velika pedijatrijska bolnica u okviru koje se nalazi nacionalni centar za lečenje dece i odraslih obolelih od cistične fibroze. U radu je okarakterisano 88 sojeva S. maltophilia od kojih 42 poreklom iz pacijenata obolelih od cistične fibroze (CF) i 46 iz pacijenata koji su imali druga obolenja (nCF). Sojevi su skupljeni u periodu od 2013. do 2015. godine u cilju izučavanja genetičke srodnosti i fenotipskih osobina. Sekvenciranje PCR proizvoda dobijenog umnožavanjem gena za 16S rRNK, elektoforeza u pulsirajućem polju (PFGE) i tipizacija sekvenciranjem više lokusa (MLST) su metode koje su korišćene za genotipizaciju. Analizirana je osetljivost na odabrane relevantne antibiotike i to, trimetoprim-sulfametaksazol (TMP/SMX), hloramfenikol, ciprofloksacin, levofloksacin i tetraciklin. Kod svih sojeva su analizirane površinske karakteristike, pokretljivost, sposobnost formiranja biofilma i adhezije na mucin. Takođe, analiziran je i uticaj različitih faktora (temperatura, pH, šejkiranje i CO2) na formiranje biofilma, kinetika formiranja biofilma kod odabranih sojeva kao i uticaj TMP/SMX na formirani biofilm. Dobijeni rezultati su statistički obrađeni u cilju utvrđivanja korelacija među njima. Sekvenciranje PCR proizvoda dobijenih umnožavanjem gena za 16S rRNK dobijeno je da svi analizirani klinički izolati pripadaju vrsti S. maltophilia sa identičnošću od 95% do 99% sa S. maltophilia iz NCBI baze podataka. PFGE analizom je potvrđeno da među izolatima postoji velika genetička heterogenost. MLST analizom je utvrđeno postojanje šest novih i tri prethodno detektovana tipa sekvence. Sojevi su bili senzitivi na sve testitrane antimikrobne agense. Takođe, kod svih sojeva je detektovana pokretljivost plivanjem, dok nijedan nije pokazivao druge tipove pokretljivosti. Među sojevima koji su pokazali sposobnost adhezije na mucin nije uočena razlika između CF i nCF izolata. Većina izolata je pokazala sposobnost formiranja biofilma (89,8%) i to podjednako zastupljeni su bili predstavnici CF i nCF grupe. Analizom formiranja biofilma u različitim uslovima rasta pokazano je da promena temperature i pH najjače utiče na formiranje biofilma i to gotovo identično na Cf i nCF sojeve. Primenom TMP/SMX u konncentraciji od 50 µg/ml dolazilo je do potpune razgradnje 24 sata starog biofilma, dok je prilikom primene 25 µg/ml taj efekat bio zavistan od soja. Kod sojeva koji su formirali jak biofilm uočena je sporija kinetika formiranja biofilma kod CF u odnosu na nCF sojeve. Šejkiranje nije uticalo na formiranje biofilma. Veliki genetički diverzitet među izolatima ukazuje da nije došlo do klonalnog širenja unutar bolnice. Pokazana je pozitivna koerlacija između pokretljivosti, formiranja biofilma i adhezije za micin. Osim toga, pozitivna korelacija je pokazana između pokretljivosti i jačine formiranog biofilma. Međutim, poređenjem fenotipskih karakteristika CF i nCF kliničih izolata uočava se da postoje razlike razlike između ove dve grupe sojeva, ali ne i CF fenotip.

Genotyping, antibiotic resistance and biofilm formation of Stenotrophomonas maltophilia clinical isolates from Serbia Stenotrophomonas maltophilia is an environmental bacterium and an opportunistic pathogen usually associated with healthcare-associated infections, which has recently been recognized as a global multidrug resistant organism. The aim of this study was genotyping and physiological characterization of S. maltophilia collected during the routine health care at The Institute for Mother and Child Health Care of Serbia “Dr Vukan Čupić“. It is the large, tertiary care pediatric hospital in Belgrade, Serbia, hosting the national reference cystic fibrosis (CF) center for pediatric and adult patients. We characterized 88 S. maltophilia strains, 42 strains of cystic fibrosis (CF) and 46 strains of non-cystic fibrosis (non-CF) origin isolated from 2013 to 2015 in order to investigate their genetic relatedness and phenotypic traits. Genotyping was performed using sequencing of 16S rRNA gene, Pulse Field Gel Electrophoresis (PFGE) and Multi locus sequencing typing (MLST) analysis. Sensitivity to five relevant antimicrobial agents was determined, namely trimethoprim/sulfamethoxazole (TMP/SMX), chloramphenicol, ciprofloxacin, levofloxacin and tetracycline. Surface characteristics, motility, biofilm formation and adhesion to mucin were tested in all strains. In addition, influence of different factors (temperature, pH, agitation and CO2) on biofilm formation, kinetics of selected biofilm producers and effect of TMP/SMX on formed biofilm were analyzed. Statistical approach was used to determine correlations between obtained results. All analyzed clinical isolates belong to S. maltophilia species with identity ranging from 95% to 99% with S. maltophilia strains from the NCBI database what was confirmed by sequencing of PCR products of amplified 16S rRNA gene. PFGE analysis confirmed that most of the isolates were not genetically related. Six new sequence types were determined and three already detected were found. Strains were uniformly sensitive to all tested antimicrobial agents. Swimming motility was observed in all strains, while none of the them exhibited swarming or twitching motility. Among strains able to adhere to mucin, no differences between CF and non-CF isolates were observed. The majority of isolates (89.8%) were able to form biofilm with almost equal representation in both CF and non-CF strains. Analysis of biofilm formation in different growth conditions showed that changing of temperature and pH had the strongest effect on biofilm formation almost equally in group of cystic fibrosis (CF) and non-CF strains. TMP/SMX in concentration of 50 µg/ml reduced completely 24 h old biofilms while concentration of 25 µg/ml effects formed biofilms in a strain dependent manner. Among strains able to form strong biofilm CF isolates formed biofilm slower than non-CF isolates. Agitation conditions did not affect biofilm formation. High genetic diversity among isolates implies the absence of clonal spread within the hospital. Positive correlation between motility, biofilm formation and adhesion to mucin was demonstrated. In addition, positive correlation between motility and strength of biofilm formed was demonstrated. Biofilm formation and motility were more pronounced among non-CF than CF isolates. However, a comparison of phenotypic characteristics of clinical isolates from CF and non-CF patients suggested that there was a difference between the two populations but not a CF phenotype.

Simbolički oblici stvaralačkog izraza u arhitektonskom delu

The symbolic forms of creative expression in a work of architecture

Pristup u ovom radu polazi od simboličke teorije kao izvora metodološkog okvira u konstituisanju tumačenja i doživljavanja arhitekture. Predmet istraživanja čine simbolički oblici stvaralačkog izraza shvaćeni kao elementi razumevanja značenja i smisla koji nosi jedno arhitektonsko delo u društvenom i profesionalnom kontekstu. Prvi oblik u kome se ispituje simbolizacija izraza u arhitektonskom delu vezuje se za značenja izražajnosti u nekom od simboličkih sistema izvan samog dela ili same arhitekture. Označen kao transcendencija, u ovom smislu stvaralački izraz pretpostavlja određene simboličke vrednosti arhitekture, bilo na nivou simboličke forme, bilo na nivou simbolike prostorne koncepcije, motivacije ili ideje arhitektonskog dela. U drugom vidu, ispitivan je oblik koji je imanentan arhitekturi kao disciplini i arhitektonskom delu kao neposredno datoj objektivnosti. U tom smislu, arhitektura je posmatrana kao neposredno data fenomenološka pojavnost u kojoj se prepliće objektivitet fizičke predstave i subjektivitet njenog doživljavanja. Koristeći opštu metodu razumevanja, u radu je primenjen postupak generalizacije na teoretskom nivou, kao i konkretna interpretacija teme (specijalizacija) u okviru metode studije slučaja, koja je sprovedena na primerima arhitektonskih realizacija arhitekte Branislava Mitrovića. U analizi izabranih primera, sa odgovarajućim komparacijama, izvedeno je tumačenje konkretnog stvaralaštva prema teoretskim aspektima teme, čime je data interpretacija kojom se dovodi u vezu konkretno tumačenje arhitektonskog dela i razumevanje stvaralaštva zasnovano na simboličkim funkcijama. Rezultat predstavlja jedan model tumačenja stvaralačkog izraza koji je, prateći datu teoretsku postavku, izložen u dva pravca: kao način transcendencije i kao način imanencije arhitektonskog dela.

This dissertation employs the symbolist theory as the methodological framework both for the interpretation and the perception of architecture. In the research the symbolic forms of creative expression are interpreted as elements of understanding the meaning of a work of architecture in a social and professional context. The meaning of expression in one of symbolic systems outside the work of architecture itself or outside architecture as such is investigated as the first form of symbolic expression of architecture. Designated as transcendence, this creative expression implies certain symbolic values of architecture, be it at the level of the symbolic form, symbolic spatial concept, motivation, or the idea of the work of architecture. The second form is discussed as immanent both to architecture as a discipline and the work of architecture as an immediately given objectivity. In that sense, architecture is observed as an immediately given phenomenon, in which the objectivity of physical representation and the subjectivity of its experience are intertwined. Implementing the general method of understanding, the dissertation employs generalization at the theoretical level, as well as the case study methodology in the concrete research issue interpretation (specialization), which is applied to the examples of architectural realizations of the architect Branislav Mitrović. In the analysis of the chosen examples, accompanied by appropriate comparisons, concrete works are interpreted in accordance with the theoretical framework of the research. In that way the concrete interpretation of a work of architecture is closely related to the creative work understanding based on symbolic functions. The resulting model of creative expression interpretation, firmly grounded in the theoretical basis, implies two ways of approaching a work of architecture: as having both transcendental and immanent qualities.

Tip i učestalost hromozomskih aberacija i značaj molekularnog markera JAK2V617F u primarnoj mijelofibrozi

Type and frequency of chromosomal aberrations and importance of molecular JAK2V617F marker in primary myelofibrosis

Primarna mijelofibroza (PMF) je hronična, maligna hematološka bolest, koja se karakteriše leukoeritroblastnom krvnom slikom, anizopoikilocitozom eritrocita u obliku suze, različitim stepenom fibroze kostne srži i hepatosplenomegalijom usled ekstramedu- larne hematopoeze. Od genetičkih specifičnosti bolesti, ističu se hromozomske aberacije u patološkim, mijeloidnim ćelijama krvi i tačkasta mutacija V617F na genu JAK2. Osnovni cilj studije bio je ispitati kariotipske i citogenetičke parametre i prisustvo mutacije JAK2V617F u genomu bolesnika sa de novo PMF. Ispitani su i drugi dijagnostički parametari, njihove međusobne korelacije i njihov uticaj na kumulativno preživljavanje bolesnika. Analiza kariotipa vršena je konvencionalnom citogenetičkom metodom. Alel-spe- cifična PCR analiza je korišćena za detekciju mutacije JAK2V617F. U studiji su korišćene deskriptivne i analitičke statističke metode. Retrospektivnom analizom citogenetičkih rezultata 61 bolesnika, registrovan je pa- tološki kariotip kod 41% bolesnika. Zastupljene su bile aberacije specifične za PMF: 13q-, 20q-, +8, ali i aberacije ređe zastupljene u ovoj bolesti. Prospektivnom studijom bilo je obuhvaćeno 144 bolesnika. Ispitana je učestalost hromozomskih aberacija u kariotipu bolesnika, učestalost mutacije JAK2V617F, njihova međusobna korelacija i korelacija sa kliničkim i hematološkim, laboratorijskim parametri- ma. Hromozomske aberacije bile su zastupljene kod 29% bolesnika. Od specifičnih abera- cija za PMF, najzastupljenija je bila trizomija hromozoma 9, a zatim 13q- i 20q-. Mutacija JAK2V617F je registrovana kod 55% bolesnika. Ispitivanjem korelacije mutacije i tipa kari- otipa, odnosno mutacije i hromozomskih aberacija različitog stepena rizika, nije registro- vana statistički značajna razlika (p=0.153). Ispitivanjem značaja kliničkih i hematološko laboratorijskih parametara registro- vana je razlika u preživljavanju bolesnika različitih prognoznih grupa primenom Lilskog, Servantesovog, IPSS, DIPSS prognoznih sistema (PS-a) (za sve p<0.001), Mayo PS za sve bolesnike (p=0.001) i Mayo PS za mlađe bolesnike (p=0.013). Ispitivanjem uticaja mutacije JAK2V617F, pokazano je da ne postoji statistički zna- čajna razlika (p=0,807) u preživljavanju bolesnika sa i bez mutacije. Ispitivanjem značaja patološkog kariotipa i pojedinih hromozomskih aberacija na preživljavanje bolesnika, primenom DIPSS citogenetičkog prognoznog sistema (CPS-a) re- gistrovana je statistički značajna razlika (p=0.004), dok primenom Lilskog, Mayo i IPSS CPS-a nije registovana statistički značajna razlika (p=0.155, p=0.214, p=0.152). Naši rezultati ukazuju da je opšta učestalost hromozomskih aberacija kod bolesnika sa PMF 32%. Najzastupljenije hromozomske aberacije su: +8, +9, 13q-, 20q-. Somatska mu- tacija JAK2V617F je zastupljena kod 55% bolesnika sa PMF. U slučaju ispitanih genetičkih parametra, nije konstatovana korelacija hromozomskih aberacija i mutacije JAK2V617F. Prognozno povoljne aberacije za bolesnike sa PMF su: solo 13q-, t/dup(1q) solo 20q-, solo +9, druge solo aberacije, dve aberacije isključujući nepovoljne i normalan kariotip. Nepo- voljne hromozomske aberacije su: kompleksno aberantan kariotip, solo +8, solo -7/7q-, solo -5/5q-, i(17q), inv(3), 12p-, 11q23, dve aberacije uključujući nepovoljne.

Primary Myelofibrosis (PMF) is a chronic, malignant hematological disease, cha- racterized by leukoerythroblastic blood picture, anisopoikilocytosis teardrop-shaped eryt- hrocyte, different degree of bone marrow fibrosis and hepatosplenomegaly due to extrame- dullary hematopoiesis. Among genetic specificities of the disease, those that stand out are chromosomal aberrations in pathological, myeloid blood cells and point mutation V617F in the JAK2 gene. The main goal of study was to examine karyotype and cytogenetic parameters and presence of JAK2V617F mutation in the genome of patients with de novo PMF. Additio- nally, other diagnostic parameters, their mutual correlations and their effect on cumulative survival rate of patients were examined. Karyotype analysis was performed by conventional cytogenetic method. Allele- specific PCR was used to detect the JAK2V617F mutation. The study used descriptive and analytical statistical methods. By retrospective analysis of cytogenetic results that included 61 patients, abnormal karyotype was registered in 41% of them. Specific PMF aberrations that were found are : 13q-, 20q-, +8, but also aberrations that are rarely present in this disease. Prospective study included 144 patients. The frequency of chromosomal aberrati- ons was tested, so as the frequency of JAK2V617F mutation, their mutual correlation and correlation with clinical and hemato-laboratory parameters. Chromosomal aberrations were present in 29% of patients. Of specific aberrations for PMF, the most common was tri- somy of chromosome 9, then 13q-and 20q-. JAK2V617F mutation was registered in 55% of patients. Examining the correlation between mutation and type of karyotype and mutation and chromosomal aberrations with various risk level, statistically significant difference was not registered (p=0.153). Examining the importance of clinical and hematological parameters, difference was registered in survival of patients with different prognostic groups applying Lille, Cervantes, IPSS, DIPSS prognostic systems (PSs) (for all p<0.001), Mayo PS for all patients (p=0.001) and Mayo PS for younger patients (p=0.013). Testing the influence of the JAK2V617F mutation, it was noticed that there is no statistically significant difference (p=0.807) in the survival of patients with and without mutations. Examining the importance of pathological karyotype and some chromosomal aberrations upon survival of patients, statistic significance (p=0.004) was registered using DIPSS cytogenetic prognostic system (CPS). Applying Lille, Mayo and IPSS CPSs, statistic significance was not registered (p=0.155 and p=0.214, p=0.152). Our results indicate that the overall frequency of chromosomal abnormalities in pa- tients with PMF is 32%. The most common chromosomal aberrations are: +8, +9, 13q- and 20q-. The incidence of JAK2V617F somatic mutation is 55% in our PMF patients cohort. Correlation between chromosomal aberrations and JAK2V617F mutation was not found. It was also concluded that prognostically favorable chromosomal aberrations for patients with PMF are: 13q-, t/dup(1q), 20q-, +9, other solo aberrations, two coexisting aberrations excluding unfavorable aberrations and normal karyotype. Unfavorable aberra- tions are: the presence of complex karyotype, +8, -7/7q-, -5/5q-, i(17q), inv (3), 12p-, 11q23, and the coexistance of two unfavourable aberrations.

Formiranje modela urbanog menadžmenta u realizaciji strateških projekata

Formation of the urban management model in the implementation of strategic projects.

Svrha i cilj doktorske disertacije je formiranje modela razvijanja novih instrumenata i principa urbanog menadžmenta u realizaciji strateških projekata u oblasti upravljanja i planiranja urbanim razvojem u promenjenim društveno-ekonomskim uslovima u Srbiji. Osnovna teza u doktorskoj disertaciji stoji na stanovištu da je za uspešnu realizaciju strateških urbanih projekata neophodno formiranje sopstvenog modela urbanog menadžmenta u upravljanju urbanim razvojem u Srbiji. Smatramo da nije moguće dosledno slediti univerzalne modele, već da mogu da se koriste pojedini principi i instrumenti pojedinačno i u kombinaciji prema mogućnostima i potrebama prilagođenim i primerenim specifičnom kontekstu. Problemski okvir istraživanja smešten je u širi društveni i ekonomski kontekst razvoja Srbije i njenih integracija sa svetom i Evropom, u kome je jedna od glavnih prepreka adekvatnog urbanog razvoja prevaziđena metodološko- teorijska postavka planerskog procesa. Stoga su neophodni novi modeli, metode i instrumenti za transformativne akcije u urbanom upravljanju, planiranju i menadžmentu strateških urbanih projekata. Novi modeli i instrumenti urbanog upravljanja i menadžmenta uzimaju u obzir logiku i funkcionisanje globalnih konkurentnih tržišta sa jedne, i održive ekonomske i društvene uticaje, potencijale i benefite za lokalitet na kojima nastaju sa druge strane. Predmet istraživanja je ispitivanje mogućnosti formiranja i primene novog modela urbanog menadžmenta u realizaciji strateških projekata u oblasti upravljanja i planiranja urbanim razvojem u Srbiji. Širi predmet istraživanja bavi se odnosom upravljanja teritorijom i proizvodnjom prostora koji stvara i pozitivne i negativne efekte. U središtu tog odnosa nalaze se strategije, mehanizmi i instrumenti za operacionalizaciju interesa društveno- ekonomskih aktera, čija upotreba zavisi od specifičnog društvenog konteksta. U istraživanju je stavljen fokus na kvalitet rešenja realizovanih strateških projekata i efekata koja ta rešenja proizvode u prostoru i na različite aktere. VII Fokus na urbani menadžment u istraživanju odnosi se na ispitivanje načina na koje akteri javnog i privatnog sektora uređuju odnose kroz različite organizacione aranžmane, zahteve koje imaju prilikom uspostavljanja pravila za saradnju, i na instrumente i resurse koje koriste u procesu planiranja i realizacije strateških projekata. Važna pitanja postaju aktivnosti, obaveze i odgovornosti koje akteri preuzimaju i rezultati koje postižu, uz analizu rizika i koristi, kako pojedinačno, tako i za zajednicu u celini. Odgovori na ova pitanja u kontekstu teorijskih i praktičnih razmatranja doprinose formiranju modela razvijanja novih instrumenata urbanog menadžmenta u realizaciji strateških projekata u promenjenim društveno-ekonomskim uslovima u Srbiji. Metodologija korišćena u istraživanju ima kvalitativni, pragmatični i multidisciplinarni karakter. Na teorijskom nivou korišćen je komparativni kritički metod za pregled i kritičku analizu teorijskih savremenih znanja u oblasti urbanog menadžmenta, upravljanja i planiranja, i to prvenstveno paradigmi pravednog grada i kolaborativno-komunikativnog pravca sa jedne, i paradigmi urbanog menadžmenta i upravljanja sa druge strane. Ishodi diskusije teorijskog i konceptualnog istraživanja su korišćenjem metode sinteze teorijskih nalaza upotrebljeni u definisanju teorijskih inputa za formiranje modela i instrumenata urbanog menadžmenta u realizaciji strateških projekata. Na pragmatičnom nivou istraživanja korišćene su odabrane studije slučajeva društveno-ekonomskih konteksta Velike Britanije i Holandije, kao i povezivanje sa kompleksnom analizom aktuelne situacije u oblasti upravljanja i planiranja urbanog razvoja u odnosu na odabranu srpsku studiju slučaja. Formiranjem modela urbanog menadžmenta u realizaciji strateških projekata i proverom njegovog funkcionisanja u Evropi i Srbiji, istraživanje nudi nove originalne nalaze za proširivanje postojećih i uvođenje novih teorijskih, metodoloških i praktičnih znanja i inovacija, kao i novih instrumenata i principa urbanog menadžmenta pozicioniranih u okvire razumevanja operacija na tržištu u oblasti urbanističkog planiranja i upravljanja urbanim razvojem u Srbiji. VIII Rezultat istraživanja na teorijskoj i praktičnoj ravni je dopuna i unapređenje postojećih teorijskih i praktičnih znanja i modela planiranja i upravljanja prostornim razvojem u Srbiji, koji su uglavnom zasnovani na racionalnoj, i u poslednjih nekoliko godina delom na kolaborativnoj paradigmi. Na taj način formirani model urbanog menadžmenta u realizaciji strateških projekata uspostavlja relacije i povezuje savremene teorije i paradigme pravednog grada i kolaborativno-komunikativnog pravca sa jedne strane, sa savremenima teorijama i paradigmama urbanog menadžmenta i upravljanja sa druge strane. Važan naučni rezultat istraživanja je identifikacija i definisanje glavnih faktora, ograničenja i uslova prilikom prilagođavanja i primene modela urbanog menadžmenta u koncipiranju i realizaciji strateških urbanih projekata u srpskom kontekstu. Primenljivost rezultata istraživanja i modela urbanog menadžmenta u realizaciji strateških urbanih projekata ogleda se u tome što lokalnim upravama u srpskom kontekstu nudi proširivanje upravljačkih i planerskih aktivnosti na čitavu paletu savremenih planerskih pristupa, metoda, alata i instrumenata urbanog menadžmenta koji su pozicionirani u okvire razumevanja operacija na tržištu. Kroz istraživanje smo potvrdili tezu da odnosi između aktera i njihova organizacija, akcije koje preduzimaju i pravila koja formiraju zajedno imaju ključnu ulogu u realizaciji velikih strateških urbanih projekata, pre nego institucionalna zakonska pravila i procedure, iako ona predstavljaju kontekst i određuju okvire u kojima se akcije i odluke donose za iniciranje, planiranje, menadžment i realizaciju strateških urbanih projekata.

The purpose and objective of doctoral dissertation is the formation of urban management model and development of new instruments and principles in the implementation of strategic projects in the field of urban governance and planning of the development in changed socio-economic conditions in Serbia. The main thesis in the doctoral dissertation argues that for the successful implementation of strategic urban projects it is necessary to form own urban management model in urban governance and development processes in Serbia. We believe that it is impossible to consistently follow universal models, but instead only use certain principles and instruments separately and in combination according to the possibilities and needs adjusted and appropriate for the specific context. The problem of the research is framed in the broader social and economic context of the development of Serbia and its integration with the world and Europe, in which one of the main obstacles of adequate urban development is obsolete methodological and theoretical construct of the planning process. Therefore, new models, methods and instruments are needed for transformative action in urban governance, planning and management of strategic urban projects. New models and instruments of urban governance and management consider the logic and the functioning of the global competitive markets on one hand, and sustainable economic and social impacts, potentials and benefits for the locality where they originate on the other side. The subject of the research is in examining possibility of formation and application of new model of urban management in the implementation of strategic projects in the field of urban governance and planning of the development in Serbia. Wider subject of the research deals with the relation of the territorial governance and the production of space, which creates both positive and negative outcomes. Strategies, mechanisms and instruments for the operationalization of interest of socio-economic actors are at the heart of this relation, and their use depends on the specific social context. The focus of the research is on the quality of created solutions of implemented strategic projects and the effects generated by those solutions within the territory and towards various actors. X Focus on urban management in the research is in examining ways in which actors in public and private sector regulate relations through different organizational arrangements, requirements they have in creating rules for cooperation, and instruments and resources they use in the planning process and implementation of strategic projects. Actions, obligations and responsibilities which actors accept and results they achieve together are becoming important issues, with the analysis of risks and benefits, for each actor and for the community as a whole. Answers to these questions in the context of theoretical and practical considerations contribute to the formation of urban management model and development of new instruments and principles in the implementation of strategic projects in changed socio-economic conditions in Serbia. The methodology used in the research has a qualitative, pragmatic and multidisciplinary character. We used the critical comparative method on a theoretical level to review and critically analyze contemporary theoretical knowledge in the field of urban management, governance and planning, primarily the just city and communicative and collaborative paradigms on one hand, and urban management and governance paradigms on the other hand. The outcomes of the discussion from the theoretical and conceptual research were used with the method of synthesis of theoretical findings in defining the theoretical input for the formation of model and instruments of urban management in the implementation of strategic projects. Selected case studies from the socio-economic context of Great Britain and the Netherlands were used on the pragmatic level of the research, as well as their connection with the complex analysis of the current situation in the field of urban governance and planning in the relation with the selected Serbian case study. By forming urban management model in the implementation of strategic projects and verification of its functioning in Europe and Serbia, the research offers new original findings for the extension of existing and introduction of new theoretical, methodological and practical knowledge and innovations, as well as new instruments and principles of urban management positioned in the framework of market operations understanding in the field of urban planning and governance of urban development in Serbia. XI The result of the research on theoretical and practical level is the addition and improvement of existing theoretical and practical knowledge and models of governance and planning the urban development in Serbia, which are mainly based on rational, and in the last few years partly on the collaborative paradigm. The urban management model in the implementation of strategic projects in this way establish relations and connects contemporary theories of a just city and communicative and collaborative paradigms on one hand, with contemporary theories of new public management, and governance organizing on the other hand. The important scientific result of the research is the identification and definition of the main factors, conditions and limitations in adapting and applying the urban management model in the design and implementation of strategic urban projects in the Serbian context. The applicability of the research results and the urban management model in the implementation of strategic urban projects is reflected in the fact that it provides broadening of management and planning activities for local governments in the Serbian context towards the entire range of modern planning approaches, methods, tools and instruments of urban management positioned in the framework of market operations understanding. We have confirmed through the research the thesis that the relations between actors and their relations, actions taken and rules they create together have the crucial role in the realization of large strategic urban projects, rather than the institutional and legal rules even though they represent a context and determine the framework in which these actions and decisions are made for initiating, planning, management and implementation of strategic urban projects.

Unapređenje prostorne organizacije stanovanja u romskim naseljima u Beogradu pod uticajem načela Romanipena

Improvement of housing spatial organization in Roma settlements in Belgrade under influence of Romanipen principles

Praksa rešavanja pitanja stanovanja socijalno ugroženih Roma proces je u nastajanju interakcijom teorijskog i praktičnog, postavljenim izvan projektantske struke u konceptu savremenog društva kao kulturno pluralnog. Nepoznanice o načinu stanovanja Roma uzrokuju nedostatak metoda i tehnika za izbor i razvoj projektovanja adekvatnog stanovanja. Namera rada je da, u ključu produbljivanja saznanja o stanovanju Roma, pokaže da nekoliko međusobno uslovljenih osnovnih karakteristika funkcionalne organizacije romske zajednice zasnovane pod uticajem načela Romanipena predstavljaju etos formiranja prostornih struktura i realizacija načina građenja u neplanski nastalim i razvijanim romskim naseljima u Beogradu. Rad najpre polazi od šireg tumačenja teritorijalizacije izraženog u stavu da između prostornog i društvenog nivoa u romskim naseljima postoji relacija koja omogućava razumevanje onoga šta ta kulturna grupa proizvodi u prostoru. Prostorne strukture romskih naselja nastale su bez uticaja struke i predstavljaju kvalitet koji je proistekao iz potreba funkcionalne organizacije zajednice i odvijanja aktivnosti svakodnevnog života kojim dominira kolektivni životni stil zasnovan na konkretnim načelima Romanipena. Predmet rada usmeren je na prepoznavanje prostornog izražavanja dva stepena kolektiviteta beogradskih Roma – porodice i zajednice – u odabranim naseljima koja prema svojoj unutarnaseljskoj strukturi predstavljaju reprezentativan uzorak. Dalji razvoj rada odnosi se na uporednu analizu rezultata istraživanja – u formi utvrđenih i sistematizovanih karaktera fizičke strukture – sa preliminarnim okvirom mogućnosti koje propisuju postojeća i važeća dokumenta kako bi se naučno opisane činjenice o romskom stanovanju izrazile i klasifikovale u upotrebnu bazu programskih načela koja omogućava praktične projektantske odgovore u različitim strategijama primene. Doprinos rada ogleda se najpre u osvetljavanju činjenice da romski mikroenvajronment nudi pravac sopstvene šanse za učestvovanjem u stvaranju savremene građene sredine. Definisanje programskih načela ovom procedurom postiže dvostruki efekat: najpre upućuje u arhitektonsko i urbanističko projektovanje uslova za način života koji je Romima prihvatljiv, a potom i omogućava projektantsku aktualizaciju strateških principa rešavanja stanovanja socijalno ugroženih Roma, a metodološki i drugih kulturno različitih grupa.

The practice of resolving the issues of socially endangered Roma housing is a process developing through the interaction of the theoratical and the practical, situated outside the design profession in the concept of the contemporary society as multicultural. The lack of knowledge on the Roma lifestyle and housing principles causes a lack of methods and techniques to choose and develop the designs for adequate housing. The intent of this paper is to present how several basic interdependent characteristics of the functional organization of Romanipen based Roma communities represent the ethos of the formation of spatial structures and construction methods in unplanned Roma settlements in Belgrade, using a deeper understanding of the Roma housing principles. Тhe paper starts with the broader interpretation of territorialization which suggests the existence of a relation between the spatial and the social level of Roma settlements; a relation that makes possible the understanding of what this cultural group produces in space. The physical spatial patterns of informal Roma settlements were created without professionals’ influence and represent a quality induced by the need to functionally organize a community where everyday activities take place. Therefore, the characteristics of spatial organization and construction methods in unplanned Roma settlements are the actual object of research when examining a collective lifestyle as the dominant pattern in everyday behavior determined by concrete Romanipen principles. The subject of the paper is focused on recognizing spatial expressions of two different degrees of Belgrade Romani collectives – the family and the community – in select settlements which constitute a representative sample due to their intra-settlement structures. Further on, the paper compares research results – a systematic overview of characters of physical structures and construction principles – with the preliminary framework of possibilities supported by existing, valid documents in order to express and classify scientifically described facts on Romani housing as an operable database of programmatic principles. The purpose of the paper is firstly to emphasize the fact that the Roma microenvironments already offer their own ways for participation in the creation of cities of the future. Defining programmatic principles through this procedure has a dual effect: it directs towards architectural and urban design of lifestyle conditions acceptable to the Romani community; also, it enables the design actualization of strategic principles for resolving housing for socially endangered Romani and, methodologically, other culturally different groups as well.

Uticaj opšte anestezije na spavanje u eksperimentalnom modelu holinergičke neuropatologije Parkinsonove bolesti - mogući rizici postoperativnog oporavka

Impact of general anesthesia on sleep in the experimental model of Parkison's disease neuropathology - possible risks of postoperative recovery

U ovoj doktorskoj disertaciji ispitivan je uticaj ketamin/diazepam i pentobarbital opšte anestezije na EEG mikrostrukturu i obrazac disanja tokom anestezije, kao i na arhitekturu spavanja i strukturu prelaznih stanja, EEG mikrostrukturu i dinamiku epizoda svih faza spavanja nakon anestezije, kako u fiziološkim kontrolama, tako i u eksperimentalnom modelu holinergičke neuropatologije Parkinsonove bolesti (bilateralna lezija PPT jedra). Rezultati ove doktorske disertacije pokazali su da je ketamin/diazepam anestezija operativnog nivoa izazvala ozbiljan poremećaj respiratornog obrasca i EEG mikrostrukture tokom anestezije. Međutim, jednako vreme potrebno za uspostavljanje stabilne anestezije u pacova sa bilateralnom lezijom PPT jedra kao i dugotrajni suprimirajući efekti na povećanu NREM beta i teta amplitudu (elektrofiziološki markeri deficita holinergičkih neurona PPT jedra) nakon anestezije, ukazuju da je ova anestezija potencijalno povoljnija, kako za ulazak u anesteziju, tako i za NREM fazu spavanja nakon operativnih zahvata kod gerijatrijskih pacijenata, kao i pacijenata obolelih od Parkinsonove i Alchajmerove bolesti. Nasuprot ketamin/diazepam anesteziji, pentobarbital anestezija ima povoljnije dejstvo na REM fazu spavanja nakon anestezije, kako kod fizioloških kontrola, tako i u eksperimentalnom modelu holinergičke neuropatologije Parkinsonove bolesti na osnovu: dugotrajnih poremećaja REM spavanja i strukture prelaznih stanja kod fizioloških kontrola koje izaziva ketamin/diazepam anestezija; suprimirajućih efekata obe anestezije operativnog nivoa na produženu REM/REM1 fazu spavanja i poremećaj njihove EEG mikrostrukture kod bilateralne lezije PPT jedra; suprimirajućeg efekta pentobarbital anestezije na povećani broj NREM/REM/NREM prelaza uzrokovanih lezijom PPT jedra; kao i na osnovu EEG mikrostrukture tokom stabilne pentobarbital anestezije, koja se ne razlikuje u poređenju PPT lezije i fiziološke kontrole.

The aim of this doctoral dissertation was to follow the impact of ketamine/diazepam and pentobarbital anesthesia on the EEG microstructure and respiratory pattern during anesthesia, and on the post-anesthesia sleep/wake states architecture and transition structure, EEG microstructure across sleep, and all the sleep state episodes dynamics in the physiological condition, as well as during the PPT cholinergic neuropathology (the experimental model of Parkinson’s disease cholinergic neuropathology). Ketamine/diazepam anesthesia induces more alterations in the EEG microstructure and respiratory pattern than does the pentobarbital anesthesia in the PPT lesioned rats. In addition, the equal time required to establish an anesthetized state in the PPT lesioned vs. control rats, and the long-term suppressive effect on augmented NREM beta and theta amplitudes (the hallmarks of PPT cholinergic neuronal loss in rat) suggest ketamine/diazepam anesthesia as potentially more beneficial both for anesthesia induction and for post-anesthesia NREM sleep in the surgical procedures of the elderly, and Parkinson’s, and Alzheimer’s patients. In contrast to the ketamine/diazepam anesthesia the pentobarbital anesthesia is much more beneficial for the post-anesthesia REM sleep in physiological conditions as well as during the PPT cholinergic neuropathology on the basis of: the long-lasting REM sleep and transition structure disorders in physiological controls, induced by the ketamine/diazepam; the abolishing effects of both anesthetic regimens on the post- anesthesia prolonged REM/REM1 sleep and on their EEG microstructure disorders in the PPT lesioned rats; the abolishing effect of pentobarbital on the increased NREM/REM/NREM transitions, caused by the PPT lesion; and the equal EEG microstructure during stable pentobarbital anesthesia in the PPT lesioned rats versus controls.

Naučna estetika arhitekture Milutina Borisavljevića

Miloutine Borissavliévitch and his scientific aesthetics of architecture

Ova doktorska disertacija predstavlja doprinos proučavanju i istraživanju estetike i teorije arhitekture. Okosnicu rada čini naučna estetika arhitekture koju je zasnovao i koncipirao srpski arhitekta i estetičar Milutin Borisavljević u drugoj deceniji XX veka. Milutin Borisavljević je bio arhitekta, teoretičar, kritičar, estetičar arhitekture i naučnik koji se bavio izučavanjem optičko- fizioloških fenomena, za sobom je ostavio bogatu zaostavštinu koja broji preko sto trideset izvedenih građevina, oko sto neizvedenih projekata, crteža i skica, osamnaest objavljenih knjiga, blizu sto sedamdeset naučnih radova, i više desetina kritičkih tekstova publikovanih u srpskim i francuskim novinama, zbornicima i časopisima. Glavni razlog za nastanak ove doktorske disertacije nalazi se u činjenici da je bogato i raznovrsno stvaralaštvo Milutina Borisavljevića do danas ostalo neistražena oblast. O Milutinu Borisavljeviću i njegovoj naučnoj estetici arhitekture nije napisana nijedna sveobuhvatna naučna studija. Njegov projektantski rad delimično je bio predmet analiza pojedinih autora u vidu kratkih studijskih tekstova, međutim niko nije u celosti istražio Borisavljevićev teorijski koncept naučne estetike arhitekture. Iako je više istraživača isticalo važnost i značaj koji pripadaju Milutinu Borisavljeviću u svetskim analima estetike, u svojoj lokalnoj sredini taj veliki doprinos estetici arhitekture je zaboravljen. Često možemo iv naići na konstatacije da Borisavljevićev rad u oblasti estetike predstavlja nešto dragoceno i veoma značajno, po čemu ceo svet pamti njegovo ime, ali nigde ne možemo naći objašnjenje šta zaista predstavlja naučna estetika arhitekture? Kakva je to oblast ili vrsta estetike? I koji su to problemi kojima se ova estetika bavi? Na ova ključna i mnoga druga pitanja nastoji da odgovori ova doktorska disertacija.

This PhD thesis represents further contribution to the study and research of aesthetics and theory of architecture. The framework of this PhD thesis is scientific aesthetics of architecture established and conceived by Milutin Borisavljevic, Serbian architect and esthetician, during second decade of 20th century. Milutin Borisavljevic, who was an architect, theorist, critic, esthetician of architecture and scientist which studied optical and physiological phenomena, left behind rich legacy consisted of more than one hundred thirty buildings, close to one hundred unrealized projects, drawings, sketches, twenty published books, over one hundred seventy scientific papers and dozens of critics published in Serbian and French newspapers, collections and magazines. The main reason for creation of this PhD lies in the fact that fullness and variety of Milutin Borisavljevic creative work to this date have remained unexplored area. No comprehensive scientific study has been written about Milutin Borisavljevic and his scientific aesthetics of architecture. His work in architectural design was partially subject of analysis of some authors in the form of short study texts, however, no one fully explored his theoretical concept of scientific aesthetics of architecture. Although several researchers emphasized global significance and importance of Milutin Borisavljevic`s work in the field of aesthetics, in the local environment, his great contribution to the aesthetics of architecture has been forgotten and not fully recognized. vi Often we come across a statement that Borisavljevic`s aesthetics theory is something valuable and very important which makes the whole world remembers his name, but nowhere is to be found what the scientific aesthetics of architecture exactly is. What kind of area is this or what kind of aesthetics? What problems fall in the area of the aesthetics in question? On this key questions and many others this PhD thesis strives to answer.

Zone zaštite srpskih manastira na Kosovu i Metohiji

Protective zones of the Serbian monasteries in Kosovo and Metohija

Manastir je jedinstvo izgrađene strukture, posvećenog života i prostora u kome se taj život odvija. Imajući uvek duhovnu misiju, manastiri mogu biti u statusu kulturnog ili prirodnog dobra, kategorisani kao dobra od nacionalnog značaja ili kao svetska baština. Na području Kosova i Metohije (u funkciji, u materijalnim ostacima ili u predanju) nalazi se preko 1500 svetih mesta (manastiri, crkve, groblja) koja svedoče istoriju i pravoslavnu duhovnost srpskog naroda vekovima unazad. Jedinstvo srpske svetovne i duhovne vlasti postignuto u periodu vladavine dinastije Nemanjića odrazilo se i u crkvenoj umetnosti – nastankom originalnih stilskih škola građenja i urbanih formi, pretežno sa građevinom hrama u centru slobodnog prostora porte, zatvorenog izgrađenom strukturom. Između svih važnih manastira na Kosovu i Metohiji, u prelaznim decenijama iz XX u XXI vek, u radu je izdvojeno petnaest manastira za koje se može reći da su od posebne važnosti za Srpsku pravoslavnu crkvu (prema sintezi u delu Zadužbine Kosova), za Repuliku Srbiju (prema statusu zaštite, pretežno su zaštićeni kao spomenici kulture), i za Crkvu i Državu istovremeno (vezano za Državne pregovore vođene 2006-2007.g. u Beču). Osim u vreme otomanske vladavine, manastiri su stradali i u ratovima u prvoj polovini XX veka, istovremeno izlagani napadima lokalnog albanskog stanovništva. Posle Drugog svetskog rata, manastirska imovina oduzeta je kroz donošenje niza zakona u duhu novog društvenog i državnog uređenja. U okolnostima okončanja NATO bombardovanja SRJ 1999.g., donošenja Rezolucije 1244 i uvođenjem protektorata međunarodne zajednice na Kosovu i Metohiji, u etnički motivisanim napadima stradalo je 129 svetih mesta, od čega 34 u takozvanom Martovskom pogromu 2004.g. Uništavanje srpskog istorijskog, duhovnog i kulturnog nasleđa potvrdile su i međunarodne misije (UNESCO, 6 Council of Europe), posle čega su manastiri Dečani, Gračanica i Pećka Patrijaršija upisani na UNESCO’s World Heritage Lists. Napadi su otvorili potrebu uspostavljanja zona zaštite oko srpskih svetih mesta, pre svega u cilju njihove bezbednosti. Za Državne pregovore u Beču 2006.g., srpski eksperti pripremili su tehničku dokumentaciju za zone zaštite na 39 složenih lokacija u kojima su sadržana 24 manastira. Usledio je Aneks V Ahtisarijevog plana za versko nasleđe, čije je stručne nedostatke u određivanju zona zaštite i upravljanju zonama srpska pregovaračka strana pokazala analizama i izložila ih na pregovorima u Beču 2007.g. To međunarodnu zajednicu nije sprečilo da podrži, iz Ahtisarijevog plana proizašlu, tzv. zakonsku regulativu o specijalnim zonama zaštite predmetnih lokacija koju su donele stručno nespremne takozvane privremene institucije samouprave Kosova. Ahtisarijev plan podržao je i model obnove srpskih hramova prema Memorandumu, koji je u dva navrata potpisala Srpska pravoslavna crkva sa istim tzv. privremenim institucijama Kosova, uz svedočenje UNMIK, gde je na čelo sprovođenja obnove stao Council of Europe. Osim ovog primera, u radu se razmatra model obnove manastira Banjske kroz čitav XX vek, prvo, kada su istraživački radovi završeni obnovom hramovne građevine i kasnije, kada je istraživanje manastira završeno konzervacijom otkrivenih ostataka građevina tj. bez oživljavanja manastira, koje će se u okviru modela održive obnove dogoditi u XXI veku. Izrada urbanističkog Plana za manastir, naselje i banju Banjska potvrdila je integrativni pristup obnovi kulturnog nasleđa u saradnji Srpske pravoslavne crkve i državnih institucija. Određivanje zona zaštite razmatrano je u svetlu crkvene regulative od Sabora u Jerusalimu 50.g., kroz Svete kanone i monaška pravila, do savremenih srpskih zakona o zaštiti kulturnih dobara i prirode, planiranju i izgradnji, crkvama i verskim zajednicama. Uzeti su u obzir i uticaji međunarodnih konvencija u oblasti zaštite kulturnog, prirodnog i nematerijalnog nasleđa, UNESCO kriterijumi o utvrđivanju univerzalnih vrednosti i UNOSEK kriterijumi korišćeni u toku Državnih pregovora u Beču. 7 Ustanovljena su tri zonska pojasa zaštite sa mogućnošću njihovog unutrašnjeg raslojavanja, svi objedinjeni Bogoslužbenom funkcijom. To su: zadate zone zaštite manastira (oltar, hram, porta, obodna izgrađena struktura), zone zaštite okruženja manastira (vlasnički obuhvat, urbanistička zona, kulturni predeo) i posebne zone zaštite (posvećen prostor i posvećeno vreme). Istraživanjem su utvrđena i merila za uspostavljanje zona zaštite kroz pitanja procedure potom i tehnička merila data za svaki zonski pojas zasebno. Određivanje i upravljanje zonama zaštite zahteva punu usaglašenost Srpske pravoslavne crkve i institucija Republike Srbije, uz usaglašenu primenu Svetih kanona, domaće regulative i stranih konvencija. U složeni proces određivanja zona zaštite manastira neophodno je uključiti stručnjake iz istorije umetnosti, arhitekture i urbanizma i posebno poznavaoce monaškog života.

A monastery is a union of a built structure, of consecrated life and the overall space in which that life unfolds. Always having a spiritual mission, monasteries may have a status of a cultural or a natural property, categorized as a property of national significance or as a world heritage site. In Kosovo and Metohija (either in function, in the form of architectural remains or as a part of the old tradition) there are about 1500 holy places (monasteries, churches and cemeteries) which bear witness to the Orthodox spirituality and history of the Serbian nation throughout the ages. The unity of the Serbian state and the Serbian church achieved during the rule of the Nemanjic dynasty has reflected in the church art, in original architectural styles and urban forms, where the church building was always in centre of the free space of the churchyard enclosed with the structures built around it. Among all of the significant monasteries in Kosovo and Metohija, in the decades between 20th and 21st centuries, there are fifteen of them that stand out as the most important monasteries for the Serbian Orthodox Church (according to the book Legacies of Kosovo), for the Serbian state (according to their protection status, mainly protected as monuments of culture) and for the Church and the State together (according to the platform for the State Negotiations held in Vienna in 2006/2007). In addition to the destruction during the Ottoman rule, the monasteries were also destroyed in the wars in the first half of 20th century, while at the same time being exposed to the attacks of the local Albanian population. After World War II, the monastery property was dispossessed by the legislation which was passed within the new social establishment of the state. Under the circumstances following the NATO bombing of Yugoslavia in 1999, the UN Resolution 1244 and the arrival of the international forces, 129 sacred places, 34 of which during the 2004 March riots, were destroyed in Kosovo and Metohija in ethnically motivated attacks. The international missions (UNESCO, Council of Europe) confirmed the destruction of the Serbian historical, spiritual and cultural heritage after which the UNESCO inscribed the 9 monasteries Decani, Gracanica, and the Patriarchate of Pec on the UNESCO’s World Heritage List. The attacks created a need to establish protective zones around the Serbian holy sites, primarily for the sake of their safety. For the purpose of the State Negotiations held in Vienna in 2006, experts from Serbia prepared a Project for the protection of 39 complex cultural sites with 24 monasteries. This was followed by Annex V of the Ahtisaari Plan for religious heritage with certain shortcomings in the establishment and monitoring of the zones, which the Serbian experts explained through analyses in the new Project for the State Negotiations held in Vienna in 2007. This did not prevent the international community from supporting the consequent so-called legislation on special protective zones enacted by the professionally unprepared provisional institutions of Kosovo. The Ahtisaari Plan also gave support to the reconstruction of the Serbian church buildings through a Memorandum, which the Serbian Orthodox Church on signed two occasions with the same provisional institutions of Kosovo, in the presence of UNMIK. The implementation of the Memorandum is led by the Council of Europe. In addition to this example, this thesis also considers the model of the reconstruction of the monastery Banjska throughout the 20th century, first when the research works ended in the restoration of the church building and later with the conservation of the architectural remains, without revitalization of the life in monastery, which will, as a model of a sustainable renewal process, take place only at the beginning of the 21st century. The preparation of the urban-planning Project for the unified monastery, the settlement and the spa of Banjska has confirmed an integrative approach to reconstruction of cultural heritage through cooperation between the Serbian Church and the Serbian state institutions. The establishment of the protective zones was considered taking into account the church rules from the Synaxis of Jerusalem in 50 A.D., through the Holy Canons of the Church and the monastic rules, to the contemporary Serbian legislation on cultural and nature heritage, urban planning and construction, and churches and religious communities. Consideration was also given to international conventions in the sphere of the protection of cultural, natural and intangible heritage, UNESCO’s criteria on determining the 10 universal values and UNOSEK criteria used in the discussions within the State Negotiations held in Vienna. Three areas of protection have been established, with a possibility of their further division, all unified within the Liturgical function. These are the unchangeable protective zones of the monastery itself (altar of the temple, the temple, churchyard, peripheral built structure or external walls), protective zones surrounding the monastery (monastery’s properties, urban areas and cultural landscape) and special protective zones (sacral space and sacral time). The research has also established the criteria for determining the protective zones through the questions of procedure and then the technical criteria for each of the three zone areas of protection individually. The establishment and management of the protective zones must be carried out with a full consent of the Serbian Orthodox Church and the institutions of the Republic of Serbia, with a harmonized application of the Holy Canons of the Church, the Serbian legislation and international Conventions. This complex process of establishing protective zones of the monastery requires an involvement of experts in the fields of the history of art, architecture and urban- planning, and especially connoisseurs of monastic life.

Mikromorfološke karakteristike Salvia amplexicaulis Lam., S. jurisicii Košanin i S. ringens Sibth. &amp; Sm. (Lamiaceae), hemijski sastav i biološka aktivnost njihovih etarskih ulja i ekstrakata

Macromorphological characteristics of Salvia amplexicaulis Lam., S. jurisicii Košanin and S. ringens Sibth. &amp; Sm. (Lamiaceae), chemical composition and biological activity of their essential oils and extracts

Ovo istraživanje je sprovedeno u cilju dobijanja podataka o mikromorfološkim, anatomskim i hemijskim karakteristikama i biološkim dejstvima tri vrste žalfija iz Makedonije (Salvia amplexicaulis, S. jurisicii i S. ringens). Uzorci navedenih vrsta potvrgnuti su mikromorfološkim, anatomskim i citološkim analizama po prvi put, a zatim je sprovedeno sveobuhvatno ispitivanje hemijskog sastava i antioksidativne, antimikrobne, citotoksične i antineurodegenerativne aktivnosti njihovih etarskih ulja i ekstrakata. Mikromorfološke, anatomske i citološke analize vegetativnih i generativnih organa odabranih biljaka izvedene su upotrebom svetlosne i elektronske mikroskopije (skenirajuće i transmisione). Analizirane vrste poseduju bifacijalne, hipostromatične ili amfistomatične listove sa stomama paracitnog tipa i kolenhimom distribuiranim uz glavni nerv na abaksijalnoj strani. S. jurisicii i S. amplexicaulis poseduju četvorougaona, a S. ringens okruglo stablo. Vegetativni i generativni organi analiziranih vrsta nose raznovrsne neglandularne i glandularne - peltatne, kapitatne i digitiformne trihome. Broj ćelija sekretorne glavice pelatnih trihoma je varirao među analiziranim vrstama. „Niske“ kapitatne trihome (Tip I) su uočene kod sve tri vrste, dok su „visoke“ kapitatne trihome (Tip II) bile prisutne samo na cvetnim delovima S. ringens. Digitiformne trihome su nađene na listovima S. ringens i S. jurisicii. Orašice S. jurisicii i S. ringens su bile sferičnog, a S. amplexicaulis prolatno-sferičnog oblika, dok je ornamentacija površine orašica sve tri vrste okarakterisana kao retikulatna. Sitnije orašice S. amplexicaulis i S. jurisicii su osluznjavale nakon 15 minuta, a krupnije orašice S. ringens tek nakon 45 minuta. S. ringens poseduje heksakolpatna, radijalno simetrična i izopolarna polenova zrna, prolatnog oblika i sa biretikulatnom ornamentacijom egzine. U bazi plodnika S. ringens su uočljive floralne nektarije u obliku prstena...

This study was carried out in order to obtain data on anatomical, micromorphological and phytochemical characteristics as well as biological activities of three sage species originating from Macedonia (Salvia amplexicaulis, S. jurisicii and S. ringens). Plant material of these species was analyzed from anatomical, micromorphological and cytological point of view for the first time. Subsequently, their essential oils and extracts were subjected to comprenhensive investigation of chemical composition and antioxidative, antimcrobial, cytotoxic and antineurodegenerative effects. Analyses of anatomical, micromorphological and cytological characteristics of vegetative and generative plant organs were performed using light microscopy, scanning and transmission electron microscopy. The leaves of analyzed species are bifacial, hypoand amfistomatic, with paracytic stomata and collenchyma distributed in the midrib on abaxial side. Stems of S. jurisicii and S. amplexicaulis are quadriangular in cross-section, while stem of S. ringens is round-shaped. Vegetative and generative organs of examined species bear a variety of non-glandular and glandular trihomes, including peltate, capitate and digitifom ones. Secretory head cells varied between different species. „Short“ capitate trihomes (Type I) had been noticed in all of species, while „long“ capitate trihomes (Type II) were present only in floral parts of S. ringens. Digitiform trihomes were found in S. jurisicii and S. ringens leaves. S. jurisicii and S. ringens have spherical and S. amplexicaulis prolate-spherical nutlets. Ornamentation of nutlets surfaces were characterized as reticulate. Mixocarpy was observed 15 minutes after wetting in smaller nutlets of S. amplexicaulis and S. jurisicii and after 45 minutes in larger S. ringens nutlets. Pollen grains of S. ringens are prolate, hexacolpate, radially simmetric, isopolar, with bireticulate egzine ornamentation...

Analiza perifitona sa veštačkih podloga Savskog jezera i jezera Vrutci

Analysis of periphyton developed on artificial substrates in the Sava Lake and the Vrutci Lake

Značaj perifitona ogleda se u velikom doprinosu primarnoj produkciji, protoku energije i kruņenju nutrijenata u vodenim ekosistemima. U Srbiji su istraņivanja perifitona u jezerima malobrojna, a veńtačke podloge za razvoj perifitona do sada nisu bile upotrebljavane u metodologiji ekolońkih studija ove zajednice. Cilj studije na Savskom jezeru (leto 2014. godine) je bio procena uticaja različitih tipova veńtačkih podloga (staklo, keramika, vrba i tisa) i dubine inkubacije na razvoj perifitona i njegovu potencijalnu primenu u biomonitoringu. Deo istraņivanja koji je sproveden na jezeru Vrutci (leto 2015. godine) imao je za cilj ispitivanje uticaja cvetanja cijanobakterije Planktothrix rubescens na perifiton. U uzorcima perifitona mereni su netaksonomski, kao i kvalitativni i kvantitativni taksonomski parametri. Rezultati su pokazali da je dubina inkubacije na 50 cm od povrńine vode, kao i staklena veńtačka podloga najbolji izbor kod potencijalnog korińćenja netaksonomskih karakteristika perifitona u biomonitoringu. Struktura celokupne, kao i zajednice silikatnih algi, istakla je prioritetnu upotrebu inertnih veńtačkih podloga (stakla i keramike) u biomonitoringu. Taloņenje kalcita je prvi put detektovano kao posledica metalimnionskog cvetanja P. rubescens. Prisustvo ove cijanobakterije negativno se odrazilo na sve merene parametere perifitona. Fenomen zajednice silikatnih algi jezera Vrutci jeste veliki diverzitet i prisustvo 19 taksona koji se nalaze na Crvenoj listi slikatnih algi Nemačke, meĎu kojima izdvajamo Achnanthidium caledonicum, koji je dominirao u većini uzoraka i Aneumastus stroesei, koji je prvi zabeleņni predstavnik ovog roda u Srbiji. Nańi rezultati potencijalno ukazuju i na vińi stepen tolerancije silikatnih algi na prisustvo P. rubescens.

The significance of periphyton in freshwater ecosystems is fundamentally reflected in processes of primary production, nutrient cycling and energy flow. In Serbia, periphyton surveys in lentic ecosystems are scarce, and artificial substrates as methodology for ecological studies of this complex community have not been applied yet. The Sava Lake part of the study (summer 2014) aimed to determine effects of substrate types (glass, ceramic, willow and yew) and incubation depths on periphyton development and it’s potential application in biomonitoring studies. Part of research that was conducted at the Vrutci Lake (summer 2015) aimed to investigate effects of Planktothrix rubescens bloom on periphyton. In periphyton samples, non taxonomic and both qualitative and quantitative taxonomic parameteres were measured. Our results suggest that when non taxonomic parameters employment in biomonitoring purposes is assumed, periphyton should be developed on glass substrate incubated in water layer up to 50 cm. Cyanobacteria and algae community structure also pronounced preferable use of inert types of artificial substrates (glass and ceramic) in biomonitoring studies. Our results represent the first report of calite precipitation induced by P. rubescens. Presence of this cyanobacterium negatively affected all measured periphyton parameters. The other phenomenon in the Vrutci Lake was diverse diatom community in periphyton and 19 diatom taxa listed in German Red List of diatoms. Among Red List taxa, Achnanthidium caledonicum, that dominated the most of samples and Aneumastus stroesei, that is the first representative of entire genus Aneumastus in Serbia are accentuated. Our results suggest the potentilly higher tolerance level of diatoms to the P. rubescens bloom.

Plan susedstva - Norme prostorne i društvene distanciranosti

Neighbourhoud plan-the norms of spatial and social distance

Dosadašnje sociološke i antropološke klasifikacije oblika društvenosti susedstvo svrstavaju u zajednice. Imajući u vidu da se u srpskom govornom jeziku za takav oblik susedstva koristi termin komšiluk, a ne susedstvo, ovaj rad je posvećen ispitivanju finijih nijansi koje razdvajaju ova dva termina i proveri pretpostavke da je susedstvo koncept gradskosti koji se odupire homogenizujućem dejstvu ideje zajednice. Pored definicija zasnovanih na odrednicama topološke blizine i onih koje susedstvo objašnjavaju kao mesto susreta u smislu načina deljenja bliskih, zajedničkih tačaka identiteta, pojam susedstva opisuje se i kao otvoreni skup ili grupna forma. Ovaj rad ispituje tu otvorenost – meru uklapanja i meru izdvajanja grupne forme na primerima susedstva u Beogradu. Cilj razmatranja koncepcija susedstva nije razgraničenje različitih teorijskih pristupa, već da se varijeteti upotrebe ovog pojma uvedu u oblast urbanizma i prošire značenje jednog pojma koji je obeležen nostalgičnim vraćanjem ideji zajedništva i romantiziranju društvenosti zasnovanoj na solidarnosti, plemenitosti i ljubavi za bližnje. Provera teze da susedstvo nije zajednica podrazumeva ispitivanje osnovnog načela zajedništva. Endrju Mason (Andrew Mason) smatra da pojam zajednice spada u „suštinski sporne pojmove” te da nije reč o konfuziji oko značenja koja nastaje usled različitog smisla koji osobe dodeljuju određenom pojmu, već je reč o neslaganju koje izvire iz same prirode pojma (Mason 2000: 18-20). Teza da susedstvo nije zajednica uzima u obzir spornost samog pojma zajednica i ispituje relaciju između ova dva pojma čije prirode izmiču konačnom određenju. Drugim rečima, susedstvo je mesto stalnog revidiranja i korekture ideje zajedništva...

Previous sociological and anthropological classifications of sociability forms place the neighbourhood among communities. Having in mind that in Serbian language for that kind of neighbourhood it is used the term vicinity (komšiluk), but not neighbourhood, this paper is studying finer nuances separating these two terms and testing the presumptions that the neighbourhood is the concept of distance which resists the homogenizing influence of the idea of the community. Other than definitions based on the determinants of topological proximity and those explaining the neighbourhood as the meeting point according to the manner of sharing near, common identity items, the neighbourhood is also described in terms of the open gathering or group form. This work researches that openness – the measure of fitting and the measure of dissociating of group form on the examples of Belgrade neighbourhoods. The aim of the reconsidering the neighbourhood concept is not the delineation of different theoretical approaches, but to introduce the varieties of use of this term in the field of urban planning and to expand the meaning of a term which has been marked with nostalgic return to the idea of community and romantization of sociability based on the solidarity, generosity and love for neighbour. The testing of the thesis that the neighbourhood is not a community understands the review of the togetherness basic principle. Andrew Mason considers that the concept of community falls into the ‘essentially disputable concepts’ wherefore the question is not about the confusion over the meaning which results from a different senses assigned by different persons to a particular term, but about the inconsistence which comes from the very nature of the term (Mason 2000: 18-20). The thesis that the neighbourhood is not a community takes into the consideration the disputation of the term community and examines the relation between these two terms whose natures evade the final determination. In other words, the neighbourhood is the place of constant revisions and corrections of the idea of community. Unlike the community on which applies the principle of entirety, the principle of openness is applying on the neighbourhood. This significant difference shows that the neighbourhood patterns are not the ways of organizing an ensemble (concept of a community of persons living together on a definite area) but a way to connect to an open gathering. In spatial term, these are not the ways of grouping or breaking the whole (collective form) but the possibilities of regrouping, measuring, inserting (joining) and separating (suspensions) – these are the ways of maintaining the gathering, namely the group forms open...

Konstrukcija, dekonstrukcija i rekonstrukcija Viminacijuma: kontekst i koncept

Construction, deconstruction, and reconstruction of Viminacium: context and concept

Osnovni predmet istraživanja doktorske disertacije je prostor antičkog grada Viminacijuma, prestonice rimske provincije Gornje Mezije, nastalog u I veku nove ere, i važnog legijskog logora, danas arheološkog nalazišta i arheološkog parka nedaleko od gradskog naselja Kostolac u Srbiji. Rad multidisiplinarnog tima koji se sastoji od istraživača Arheološkog instituta i Centra za nove tehnologije Viminacijum iz Beograda, započet početkom ovog veka, zaustavio je pljačkanje antičkog grada, kontinuiranim istraživanjama omogućio ubrzanu prezentaciju antičkih građevina i dolazak prvih turista, a upostavljena je i zakonska granica između arheološkog nalazišta i napredujućeg površinskog kopa uglja. Međutim, u nastavak procesa zaštite i prezentacije antičkog Viminacijuma trebalo bi uključiti i uvođenje sveobuhvatnog koncepta zaštite i prezentacije prostora čiji je Viminacijum deo - koncepta kulturnog predela, uz posmatranje doprinosa svih prirodnih, istorijskih, kulturnih i društvenih činilaca Tako koncept nastao kroz rad na ovoj disertaciji ističe specifični predeo koji ne čine samo fizički ostaci građevina, već i nematerijalne vrednosti nastale kao rezultat uticaja različitih konteksta kroz evoluciju, odnosno konstrukciju, dekonstrukciju i rekonstrukciju - građevina, naselja i prostora. Ovim pristupom se prate savremene doktrine zaštite nasleđa u kojima je pojam kulturnog predela sve značajniji. Istraživanje obuhvata današnji prostor zaštićenog arheološkog nalazišta Viminacijum, ali i okolinu koja je neodvojiva od njega, u privredno- ekonomskom, kulturno-istorijskom i društveno-sociološkom smislu, kao rezultat njihove međusobne veze razvijane kroz istoriju ljudskog prisustva na njemu. Zato je bilo neophodno izvršiti istraživanje, razumevanje i valorizaciju odlika elemenata iz svih istorijskih perioda šireg područja Viminacijuma, što je kao rezultat dalo predlog novog koncepta, u kome je ono predstavljeno kao ii jedinstveni kulturni predeo. Ostala arheološka nalazišta, srednjovekovne tvrđave, manastiri, stare tradicionalne kuće, industrijsko nasleđe stvarano u poslednjih skoro sto pedeset godina, i reka Dunav, prikazani su kroz prihvatanje njihovog značaja kao ravnopravnih činilaca prostora. Pokazano je da diskontinuiteti postojanja fizičkih elemenata prošlosti ne znače i prekide u tradicijama i osnovnom ljudskom odnosu prema prirodi, jer su nematerijalne vrednosti ono što fizičke ostatke iz različitih istorijskih perioda međusobno povezuje na ovom jedinstvenom prostoru.

The main subject of this doctoral dissertation research is the area of the ancient city of Viminacium, the capital of the Roman province of Upper Moesia, founded in the 1st century AD, and an important legionary fortress, today an archaeological site and archaeological park not far from the town of Kostolac in Serbia. The work of a multidisciplinary team consisting of researchers from the Archaeological Institute and the Center for New Technologies Viminacium, from Belgrade, started at the beginning of this century, prevented the looting of the ancient city, enabled the continuous excavations of ancient buildings and the arrival of the first tourists, and established a legal boundary between the archaeological site and the progressive surface coal mine. However, the further development of the protection and presentation of the ancient Viminacium should include the introduction of a comprehensive concept of the protection and presentation of the space to which Viminacium belongs - the concept of a cultural landscape - while observing the contributions of all of the natural, historical, cultural and social factors. Thus, the concept created in this dissertation emphasises the specific landscape that is not only made up of the physical remains of buildings but also of immaterial values created as a result of the influence of different contexts through evolution, that is, the construction, deconstruction, and reconstruction of buildings, settlements and spaces. This approach is followed by modern heritage protection doctrines in which the notion of a cultural landscape is becoming more and more important. The research covers today's area of a protected archaeological site, but also an environment that is inseparable from it, in the economic, cultural, historical, social and sociological sense, as a result of their interrelationships, developed through the history of a human presence on it. Therefore, it was necessary to research, understand, and valorise the elements from all historical periods of the wider Viminacium area, which resulted in the proposal of a new concept, in which it was presented as a unique cultural landscape. Other archaeological sites, medieval fortresses, monasteries and old iv traditional houses, along with the industrial heritage created in the last nearly 150 years, and the Danube River, were analysed through their acceptance as equal factors of the area. It was demonstrated that the discontinuities of the physical elements of the past do not represent interruptions in traditions and the basic human relationship with nature, because the intangible values are links that mutually connect the physical remains from different historical periods in this unique space.

Analiza subjediničnog sastava i regulacije izoformi glutamin-sintetaze kod biljaka Arabidopsis thalliana (L.) Heynh. i Lotus corniculatus L.

Analysis of the subunit composition and regulation of glutamine synthetase isoforms from Arabidopsis thalliana (L.) Heynh. и Lotus corniculatus L.

Glutamin-sintetaza (GS) katališe asimilaciju amonijum-jona poreklom od redukcije nitrata, fotorespiracije, katabolizma aminokiselina, fiksacije N2 kod leguminoza i drugih metaboličkih procesa kod biljka. Kod viših biljaka, mala familija gena kodira različite citosolne (GS1) izoforme, dok jedan gen kodira izoformu lokalizovanu u plastidima (GS2). Zajedno sa glutamat-sintazom (GOGAT), GS je deo dvoenzimskog ciklusa odgovornog za biosintezu glutamata, koji je donor amino-grupe aminokiselinama u reakcijama katalisanim različitim transaminazama. Sticanje dodatnih saznanja o GS je od presudnog značaja za razumevanje procesa asimilacije azota kod viših biljaka, što može dovesti do bolje efikasnosti u korišćenju dostupnog azota i smanjenoj upotrebi veštačkih đubriva. U prezentovanom istraživanju ispitivana je struktura i regulacija GS izoformi iz dve model-biljke, Arabidopsis thaliana (L.) Heynh i Lotus corniculatus L. Jedan od ciljeva bio je ispitivanje mogućnosti različitih GS1 proteina A. thaliana, čiji genom kodira pet GS1 subjedinica (GLN1;1-1,5), da formiraju heteromere. Kako bi se ispitao subjedinični sastav GS1 izoformi A. thaliana, upoređeni su elektroforetski profili GS izoformi različitih GS1 SALK i SAIL knockout mutanata. Zaključeno je da GLN1;1 i GLN1;3, kao i GLN1;2 i GLN1;3, a moguće i GLN1;1 i GLN1;2 subjedinice mogu da se kombinuju u svim stehiometrijskim odnosima formirajući funkcionalne dekamerne enzime. Pored toga, kod knockout mutanata u GLN1;2 i GLN1;3 detektovana je povišena količina GLN1;1 transkripata što upućuje da GLN1;1 delom kompenzuje nedostajuće izoforme. Drugi cilj ovog istraživanja bio je ispitivanje regulacije ekspresije GS i GOGAT gena A. thaliana biljnim regulatorima rastenja: kinetinom, abscisinskom kiselinom, giberelinskom kiselinom i 2,4-dihlorfenoksiacetatom. Ekspresija GS i GOGAT gena je diferencijalno regulisana regulatorima rastenja u listu i korenu, a dobijeni obrasci ekspresije upućuju na mogući fiziološki kontekst hormonalne regulacije ovih gena tokom razvića biljke i kao odgovoru na uslove sredine. Treći cilj je bio ispitivanje hormeze indukovane fosfinotricinom (PPT) kod zvezdana (L. corniculatus). PPT je inhibitor GS koji je pronašao primenu kao neselektivni herbicid. Inhibicija GS u biljci dovodi do nagomilavanja amonijaka, nedostatka glutamina i konačno, do smrti. Utvrđeno je da biljke L. corniculatus tretirane širim opsegom koncentracija PPT-a, pokazuju dvofazni odgovor koji obuhvata stimulaciju produkcije biomase pri koncentracijama nižim od 50 μM, i inhibiciju rasta i smrt biljke pri višim koncentracijama. Stimulacija rasta niskim koncentracijama PPT-a je posledica aktivacije GS2 izoforme, dok je inhibicija rasta i smrt biljke pri visokim koncentracijama herbicida prouzrokovana inhibicijom GS1 i GS2 izoformi. Predložen je detaljan molekulski mehanizam koncentraciono-zavisne interakcije GS holoenzima sa PPT-om, koji je u skladu sa eksperimentalnim i literaturnim podacima.

Glutamine synthetase (GS) is the key enzyme involved in the assimilation of ammonia derived from nitrate reduction, photorespiration, amino acid catabolism, N2 fixation in legumes, and other metabolic processes in plants. A small gene family encodes for different cytosolic (GS1) isoforms in higher plants, while a single gene encodes for the plastidic isoform (GS2). GS operates with glutamate synthаse (GOGAT) in a two enzyme cycle. The net outcome of this cycle is the production of glutamate, which, through the action of aminotransferases, is used to synthesize other amino acids. Gaining further knowledge about GS is essential for understanding nitrogen assimilation in higher plants, which can lead to better nitrogen use efficiency and lowering of fertilizer input. The structure and regulation of GS isoforms from two model plants, Arabidopsis thaliana (L.) Heynh and Lotus corniculatus L. were investigated in the presented study. Since the A. thaliana genome encodes five GS1 subunits (GLN1;1-1,5), one of the objectives was to investigate whether different GS1 proteins from this plant are able to form heteromeric enzymes. In order to identify the subunit composition of A. thaliana GS1 isoforms, electrophoretic profiles of GS isoforms from GS1 SALK and SAIL knockout mutants were compared. It was concluded that GLN1;1 and GLN1;3, as well as GLN1;2 and GLN1;3 and possibly GLN1;1 and GLN1;2 are able to form functional heterodecamers in all stoichiometric proportions. Furthermore, in the knockout mutants lacking GLN1;2 and GLN1;3, higher GLN1;1 transcript levels were found implicating that GLN1;1, at least in part, compensates for the missing isoforms. The second objective of this study was to investigate the regulation of expression of A. thaliana GS and GOGAT genes by plant growth regulators: kinetin, abscisic acid, giberelic acid and 2,4-dichlorophenoxyacetic acid. The expression of GS and GOGAT genes is differentially regulated by growth regulators in the leaf and root. The observed patterns of expression provide insights into the hormonal regulation of these genes during development and as response to environmental cues. The third objective was to investigate phosphinothricin (PPT) induced hormesis in L. corniculatus. PPT is a potent GS inhibitor used as a non selective post emergence herbicide. GS inhibition in plants causes ammonia accumulation, glutamine depletion and eventually death. However, the growth response of L. corniculatus plants immersed in solutions with a broad range of PPT concentrations is biphasic, with pronounced stimulating effect on biomass production at concentrations ≤ 50 μM and growth inhibition at higher concentrations. The growth stimulation at low PPT concentrations is a result of activation of GS2, while the growth suppression is caused by inhibition of both GS1 and GS2 at higher PPT concentrations. A detailed molecular mechanism of concentration-dependent interaction of PPT with the GS holoenzymes from L. corniculatus is proposed. The mechanism is in concurrence with all experimental and literature data.

Pramenovi krivih trećeg i četvrtog reda dobijeni preslikavanjem pramenova konika

The pencils of curves of the third and fourth order obtained by mapping the pencils of conics

Harmonijskom simetrijom, kao potpuno bijektivnom i konformnom centralnom inverzijom, prikazana je transformacija svih tipova pramenova konika i njihovih specifičnosti u pramenove krivih trećeg i četvrtog reda i njihove preslikane specifičnosti, i obrnuto. Prepoznavanjem ekvivalentnosti inverzije sa klasičnom osnom simetrijom (nekomformna simetrija) ukazano je na neograničene mogućnosti za preslikavanje krivih i površi i dobijanje novih oblika koji će biti od koristi i u teoriji geometrije kao i u arhitektonskoj praksi

Harmonic symmetry, which is a completely bijective and conformal central inversion, is used to present the transformation of all types of pencils of conics and their specific features into the pencils of curves of the 3rd and 4th order and their mapped specific features and vice versa. The recognition of the equivalence of inversion with the classical axial symmetry (non-conformal symmetry) has created numerous possibilities for mapping curves and surfaces and obtaining new forms which can be of great use both in the theory of geometry and in the practice of architecture.

HER2 and c-MYC mutational status and INK4a/ARF methylation status in tumors, tumor margins and unaffected oral mucosa of patients with oral squamous cell carcinoma

Mutacioni status HER2 i c-MYC gena i metilacioni status INK4a/ARF lokusa u tumoru, tumorskoj margini i neizmenjenoj oralnoj sluzokoži skvamocelularnog karcinoma usne duplje

Razviće malignih oboljenja uslovljeno je akumulacijom genetičkih i epigenetičkih promena. Oralni skvamocelularni karcinom (OSCK) je najčešći malignitet usne duplje. Patohistološkim metodama evaluacije rizika od pojave recidiva i metastaza nedostaje senzitivnost, a taj problem može potencijalno da bude prevaziđen analizom molekularnih promena u OSCK-u i njegovim marginama. Ciljevi: (a) utvrditi prisustvo amplifikacije onkogena (c-MYC i HER2) i metilacije tumor supresorskih gena (P14 i P16) u tumoru, margini i zdravoj oralnoj sluzokoži pacijenata sa OSCK; (b) ustanoviti postojanje asocijacije između molekularnih i kliničkih parametara. Materijal i metode. DNK je izolovana iz tkiva tumora, margina i oralne sluzokože 40 pacijenata sa OSCK-om, operisanih na Klinici za maksilofacijalnu hirurgiju Stomatološkog fakulteta. Prisustvo amplifikacije C-MYC i HER2 gena određeno je metodom PCR u realnom vremenu, a P14 i P16 metilacije metodom metil-specifičnog PCR-a. Statistička analiza SPSS paketom je primenjena za procenu asocijacije između molekularnih i kliničkih nalaza. Rezultati: Najveća učestalost promena pokazana je u tumorskom tkivu, a najmanja u zdravoj oralnoj sluzokoži. Višestruke promene (ko-alteracije) su bile znatno češće u tumorima i tumorskim marginama nego u sluzokoži (P <0.001 odnosno P = 0.027). Amplifikacija HER2 gena u tkivu margina (P <0.001) i sluzokoži (P = 0.013), kao i postojanje tri ko-alteracije u marginama i neizmenjenoj oralnoj sluzokoži korelisane su sa kraćim preživljavanjem (P = 0.035 odnsono P = 0.027). Zaključak: HER2 amplifikacija, kao i prisustvo tri ko-alteracije u marginama i zdravoj oralnoj sluzokoži pokazale su se kao prediktori lošeg ishoda u OSCK-u.

The development of malignant diseases is due to the accumulation of genetic and epigenetic changes. Oral squamous cell carcinoma (OSCC) is an aggressive and very common malignancy of the oral cavity. Patho-histological methods lack sensitivity in terms of the evaluation of the risk of OSCC recurrence and metastases. This issue can potentially be overcome by assessing molecular changes in OSCC and its margins. Aims: (a) to determine the presence of oncogene amplification (c-MYC and HER2) and tumor suppressor gene methylation (P14 and P16) in tumor, tumor margin and healthy oral mucosa of patients with OSCC; (b) establish a potential association between molecular and clinical parameters. Material and methods: DNA was isolated from tumor, margin and oral mucosa tissue of 40 patients with OSCC, operated at the Clinic for Maxillofacial Surgery, School of Dental Medicine. The presence of C-MYC and HER2 gene amplification was determined by real-time PCR, and P14 and P16 methylation by methyl-specific PCR. Statistical analysis with SPSS was applied to estimate the association between molecular and clinical findings. Results: Tumor tissues showed the highest prevalence of alterations and oral mucosa the lowest. Multiple alterations were significantly more frequent in tumors and tumor margins compared to unaffected oral mucosa (P<0.001 and P=0.027, respectively). HER2 amplification in margin tissue (P < 0.001) and swabs (P = 0.013), as well as the existence of three co-alterations in margins and unaffected oral mucosa were correlated with shorter survival (P = 0.035 and P=0.027, respectively). Conclusion: HER2 amplification, as well as the presence of three co-alterations in margins and unaffected oral mucosa proved to be markers of poor outcome in OSCC.

Upoređivanje kvaliteta ambijentalnog osvetljenja izvedenog primenom metal-halogenih i LED izvora svetlosti

Comparison of the quality of ambient lighting realized by metal-halide and LED light sources

Osvetljenje presudno utiče na doživljaj ambijenta u noćnim satima. Do nedavno, kvalitetno ambijentalno osvetljenje podrazumevalo je upotrebu metal-halogenih izvora svetlosti sa keramičkim gorionikom, jer su njihove suštinske prednosti u odnosu na ostale klasične izvore svetlosti koji se koriste u urbanom osvetljenju (toplo-bela boja svetlosti i odlična reprodukcija boja) doprinosile adekvatnom prikazu (isticanju) karakteristika ambijenta i stvaranju prijatne atmosfere. Poslednjih godina, kao idealno rešenje za ambijentalno osvetljenje promovišu se svetiljke sa LED izvorima svetlosti. Njihove male dimenzije, usmereni svetlosni snopovi i mogućnost promene intenziteta i boje svetlosti čine ih izuzetno pogodnim za arhitektonsko osvetljenje. Međutim, postavilo se pitanje da li su LED svetiljke pogodne za ambijentalno osvetljenje, imajući u vidu karakteristike urbanih ambijenata koje je neophodno istaći i efekte koje je potrebno postići. Dok se provera fotometrijskih, energetskih i ekonomskih pokazatelja primene LED svetiljki može izvršiti merenjima ili proračunima, ostale relevantne karakteristike LED svetiljki, poput boje svetlosti, sposobnosti reprodukcije boja, subjektivnog doživljaja blještanja, osećaja bezbednosti, osećaja opuštenosti u osvetljenom prostoru i mogućnosti isticanja elemenata i detalja u prostoru, mogu se proveriti samo uz pomoć subjektivnog suda ispitanika o svakom od navedenih aspekata osvetljenja. Zbog toga ova doktorska disertacija obuhvata ne samo proveru objektivnih (energetskih, fotometrijskih i ekonomskih), nego i subjektivnih pokazatelja kvaliteta ambijentalnog osvetljenja, uz napomenu da je provera subjektivnih pokazatelja izvršena u okviru pilot projekta realizovanog na šetališnoj stazi u jednom od beogradskih parkova. v U radu su predstavljeni ne samo faktori kvaliteta ambijentalnog osvetljenja, nego i karakteristike urbanih ambijenata koje treba istaći osvetljenjem. Uz to, na osnovu analiziranih primera izvedenih ambijentalnih osvetljenja trgova, parkova i šetališta, definisane su smernice za ambijentalno osvetljenje. Analizom rezultata istraživanja sprovedenih na skoro svim kontinentima, u okviru kojih je sa raznih aspekata vršeno poređenje konvencionalnih i LED izvora svetlosti, definisani su opšti uslovi za izvođenje pilot projekata u okviru kojih se vrši poređenje različitih tipova ambijentalnog osvetljenja. U skladu sa tim, realizovan je pilot projekat u okviru koga je (uz pomoć ankete u kojoj je učestvovalo 112 ispitanika) izvršeno poređenje subjektivnih doživljaja ispitanika po pitanju relevantnih karakteristika ambijentalnog osvetljenja izvedenog pomoću LED i metal- halogenih svetiljki. Rezultati ankete su pokazali da su ispitanici smatrali da je instalacija sa svetiljkama sa konvencionalnim metal- halogenim izvorima svetlosti bolja po svim aspektima. Verovatno objašnjenje dobijenih rezultata sadržano je u činjenici da se, i pored stalnog unapređenja karakteristika LED čipova, u njihovom spektru i dalje izdvaja zračenje velikog intenziteta u području plave boje, zbog čega se primenom metal-halogenih izvora svetlosti (koje karakteriše približno kontinualan spektar) postižu bolji rezultati koji se odnose na boju svetlosti i reprodukciju boja, odnosno na ugodnost ambijenta. Napominjemo da su za osvetljenje prostora u kojima se ljudi duže zadržavaju ključni faktori kvaliteta adekvatna boja svetlosti, veoma dobra reprodukcija boja i nizak stepen blještanja. Statističkom obradom podataka primenom t-testa za jednakost srednjih vrednosti koji važi za slučaj zavisnih uzoraka, ustanovljeno je da rezultati koji se odnose na sva pitanja iz anketnog upitnika (sve aspekte vi ambijentalnog osvetljenja) mogu da se smatraju statistički vrlo značajnim ili statistički značajnim. Izvršena je i obrada rezultata ankete uz uvažavanje težinskih faktora, koja je pokazala da je u posmatranom slučaju uticaj težinskih faktora bio zanemarljiv. U cilju upoređivanja objektivnih (energetskih i ekonomskih) pokazatelja kvaliteta ambijentalnih osvetljenja izvedenih pomoću LED i metal- halogenih svetiljki, izvršeni su fotometrijski proračuni koji su se odnosili na osvetljenje šetališnih staza raznih svetlotehničkih klasa (svako od analiziranih svetlotehničkih rešenja ispunilo je sve fotometrijske zahteve koji se odnose na odgovarajuću svetlotehničku klasu). Poređenjem energetske efikasnosti osvetljenja razmatranih šetališnih staza (ukupne instalisane snage, odnosno utrošene električne energije), konstatovano je da se primenom LED umesto metal-halogenih svetiljki mogu postići uštede električne energije do 35.5%, ali i da postoje situacije kod kojih je povećana potrošnja električne energije (do 31.5%). Uz to, primenom opšte prihvaćene metode aktuelizacije troškova, izvršeno je ekonomsko poređenje fotometrijski uporedivih instalacija osvetljenja šetališnih staza. Analiza je pokazala da su pojedina LED svetlotehnička rešenja bila do 29.2% jeftinija, ali i da su preovladavala skuplja LED svetlotehnička rešenja (procentualna razlika između njihove ukupne aktuelizovane cene i ukupne aktuelizovane cene uporedivih metal- halogenih svetlotehničkih rešenja iznosila je do 45.9%). Imajući u vidu tehnologiju proizvodnje svetlosti od strane LED čipova i činjenicu da brz razvoj LED tehnologije sa aspekta energetske efikasnosti nije praćen značajnim poboljšanjima spektra emitovane svetlosti, ne očekuje se da u skoroj budućnosti LED tehnologija dostigne kvalitet spektra koji karakteriše metal-halogene izvore svetlosti. Zbog toga smatramo da će primena metal-halogenih izvora svetlosti još dugo biti opravdana u ambijentalnom osvetljenju. vii

Lighting has the crucial influence on the impression of ambience during the night. Until recently, high-quality ambient lighting was realized by the use of metal- halide lamps with a ceramic discharge tube, because their essential advantages (warm-white colour of light and excellent colour rendering) compared to other conventional light sources used in urban lighting contribute to adequate presentation of ambient characteristics and create pleasant atmosphere. During the last few years, LED lamps are promoted as the ideal solution for ambient lighting. Their small size, controlled light beam and the possibility of changing both the intensity and colour of light make them particularly suitable for architectural lighting. However, the question was raised whether LED lamps are also appropriate for ambient lighting, taking into account the characteristics of urban ambiences which should be emphasized and the effects to be achieved. While photometric, energy and economic indicators of the use of LED luminaires can be evaluated performing measurements or calculations, their other relevant characteristics, like colour appearance, colour rendering, subjective impression of glare, the sense of security and feeling of relaxation in an illuminated space, as well as the possibility of emphasizing elements and details, can only be assessed through an analysis of subjective impressions given by a number of respondents on each of these lighting aspects. Taking the above into account, the doctoral dissertation includes not only the verification of the objective (photometric, energy and economic) aspects, but also the evaluation of the subjective indicators regarding the quality of ambient lighting. The assessment of the subjective indicators was carried out through a pilot project realized on a pedestrian lane in a Belgrade park. In the dissertation, not only the ambient lighting quality factors, but also the characteristics of urban ambiences which should be emphasized by lighting, are ix presented. In addition, guidelines for ambient lighting are determined, based on the analyzed examples of the lighting solutions of squares, parks and pedestrian lanes. The analysis of the results of recent studies conducted on different continents, in which conventional and LED lighting installations were compared regarding various aspects of ambient lighting, helped to define a set of general requirements for such pilot projects. Accordingly, a pilot project, which involved 112 participants, dealt with the comparison of the subjective impressions of the respondents regarding the relevant characteristics of ambient lighting installations realized with LED and metal-halide luminaires. The results showed that the respondents found the conventional lighting installation better regarding all of the analyzed aspects. A possible explanation of the obtained results is that the LED spectrum, although continuously improving, still possesses an intensive radiation in the blue region, causing the use of metal-halide lamps (characterized by a practically continuous spectrum) more appropriate regarding both the colour appearance and colour rendering. Note that these lighting aspects, together with the elimination of glare, represent the key quality factors for the illumination of spaces where people stay for long periods of time. By statistical analysis of the obtained data, performed using the t-test for equality of means related to the case of dependent samples, it was concluded that the results for all of the questions from the survey could be considered statistically very significant or statistically significant. Also, an analysis of the survey results which included weighting factors showed that their effect is negligible in this case. In order to compare the objective (energy and economic) indicators of the quality of ambient lighting installations realized by LED and metal-halide luminaires, photometric calculations regarding the illumination of pedestrian lanes of various lighting classes were carried out (each of the analyzed lighting solutions fulfilled all x photometric requirements imposed by the corresponding lighting class). Comparing the energy efficiency of the pedestrian lane lighting (the total installed power, i.e. the consumed electricity), as the objective indicator, it was concluded that energy savings of up to 35.5% could be achieved installing LED instead of metal-halide luminaires, as well as that there are situations in which electricity consumption was increased (up to 31.5%). In addition, by applying the generally accepted cost-discount method, the comparable lighting installations were economically evaluated. The analysis showed that some LED lighting solutions were up to 29.2% cheaper, but that expensive LED lighting solutions prevailed (the difference between the total actualized costs of the comparable LED and metal- halide lighting installations amounted up to 45.9%). Due to the technology of LED light production, as well as the fact that the fast development of the LED technology in terms of energy efficiency is not accompanied by significant improvements of the light spectrum, it is not expected that the LED technology will reach the spectrum quality of metal-halide lamps in the near future. Therefore, we believe that the application of metal-halide light sources will be justified in ambient lighting for years.

Sinergistički efekat genomskog i sredinskog stresa u dve populacije Drosophila subobscura

The synergistic effect of genomic and environmental stress in two populations of Drosophila subobscura.

Jedinke uzorkovane iz dva ekološki različita staništa, i hibridi nastali njihovim ukrštanjem (unutar-linijski, unutar-populacioni i među-populacioni) izlagane su povećanoj koncentraciji olova, čime je omogućeno da se utvrdi razlika u odgovoru jedinki različitog nivoa heterozigotnosti genoma na sredinski stres odnosno, testira hipoteza o heterozigotnoj superiornosti u stresnim uslovima i utvrdi populaciono specifičan odgovor na prisustvo olova. Sinergistički efekat stresa evaluiran je na tri nivoa: populacionom - analizom komponenti adaptivne vrednosti (dužina razvića i preživljavanje od jaja do adulta), morfološkom – analizom varijabilnosti veličine krila i biohemijskom - praćenjem ekspresije proteina toplotnog šoka. Rezultati su pokazali redukciju svih ispitivanih osobina kod inbredovanih jedinki i njihovo povećanje kod jedinki sa višim nivoom heterozigotnosti genoma, potvrđujući hipotezu heterozigotne superiornost koja se intenzivira u stresnim uslovima. Uočen je trend sinergističkog efekta genomskog i sredinskog stresa za komponente adaptivne vrednosti, koji je statistički značajan za veličinu krila, s obzirom da je najintenzivnija redukcija primećena kod inbredovanih jedinki u stresnim uslovima. Nije dobijen populaciono specifičan odgovor u prisustvu olova, ali je utvrđeno da na ishod međupopulacione hibridizacije u stresnim uslovima veliki uticaj ima poreklo ženke i evoluciona istorija njene prirodne populacije. Može se zaključiti da je održavanje genetičke varijabilnosti od velike važnosti za opstanak populacija, i da čak i nizak stepen inbridinga utiče na pad adaptivne vrednosti, naročito u stresnim uslovima sredine.

The subject of this dissertation is population genetic analysis of synergistic effect of genomic and environmental stress in model species Drosophila subobscura. Individuals sampled from two ecologically distinct habitats, and their hybrids with different levels of genome heterozygosity (intra-line, intra-population and inter-population hybrids) were exposed to elevated concentrations of lead, aiming to test hypothesis of heterozygote superiority in stressful conditions and detect population specific response to lead pollution. Synergistic effect of stress was evaluated at three levels: population - through analysis of fitness components (development time and egg-to-adult survival), morphological - through analysis of variability in the wings size and biochemical – through the expression of heat shock protein. The results showed a reduction of all investigated traits in inbred individuals and their increase in those with higher levels of genome heterozygosity, confirming the hypothesis of heterozygote superiority which intensifies under stressful conditions. Analysis of fitness components indicates synergistic effect of genomic and environmental stress, and variance in wing size shows statistical significance, since the greatest reduction of all traits was observed in inbred groups. Population specific response to lead pollution was not detected, but significant influence of female origin on the outcome of between- population hybridization under stressful conditions was observed. It can be concluded that the maintenance of genetic variation is important for the survival of populations in changing environments, and even low levels of inbreeding lead to decline in adaptive value, especially in stressful environments.

Model teritorijalnih informacionih sistema za podršku održivom urbanom razvoju Srbiji

Territorial information system model to support sustainable urban development in Serbia

Potreba za informacijama raste na svim nivoima, od nivoa donosioca odluka višeg ranga na nacionalnom i internacionalnom nivou, sve do najnižih individualnih nivoa. Jedan od velikih savremenih izazova je razvoj informacione podrške održivom razvoju. Napori se fokusiraju na razvoj IKT podržanih alata zasnovanih na GIS tehnologijama, koji obezbeđuju efikasno skupljanje, skladištenje, ažuriranje, rukovanje, analiziranje i prikazivanje svih oblika podataka i informacija vezanih za teritoriju, a koji su neophodni za razumevanje teritorije u procesu kolektivnog donošenja odluka i kreiranju održivih politika i planova. U održivom razvoju, svako je korisnik i izvor informacije. Integralni pristup održivom razvoju zahteva unapređenje i široku primenu savremenih metoda, alata, tehnika i procedura planiranja i upravljanja, kako bi se obezbedila najbolje moguće razumevanje fenomena loklane tertorije u u grupnom procesu donošenja odluka o specifičnim akcijama i oceni njihovog uticaja, rizika i koristi na životnu sredinu, socijalni i ekonomski razvoj. Ovo istraživanje motivisano je potrebom stvaranja uslova za efikasno uključenje Srbije u aktuelne svetske razvojne tokove. U ovom trenutku, aktivnosti na lokalnom i regionalnom nivou širom sveta u oblasti unapređenja informacione podrške integralnom teritorijalnom razvoju fokusirane su na razvoj i primenu teritorijalnih informacionih sistema zasnovanih na IKT i GIS tehnologijama, sa ciljem postizanja veće efikasnosti uprave, kvalitativnog unapređenja vertikalne i horizontalne koordinacije, saradnje i racionalizacije procesa u oblasti informacionih i komunikacionih aktivnosti. Istraživanje svetskih iskustava u oblasti razvoja i primene ovakvih koncepata doprinosi razumevanju mogućnosti i ograničenja njegove primene u Srbiji. Predmet ovog istraživanja je razvoj modela teritorijalnih informacionih sistema za podršku održivom urbanom razvoju u Srbiji, kao novog instrumenta komunikacije i alata za podršku odlučivanju u procesu kolaborativnog planiranja i upravljanja razvojem na lokalnom nivou. Istraživanje je usmereno ka sturktuiranju modela koji bi bio aplikativan u aktuelnim uslovima procesa promena planerske paradigme i prakse planiranja, ali i u budćnosti kada se očekuje uspostavljanje novog modela planiranja zasnovanog na aktivnom učešću svih aktera razvoja. Primena ovakvog modela u Srbiji trebalo bi da, sa jedne strane, doprinese procesima promena i unapređenja u oblasti sistema planiranja urbanog i prostornog razvoja, a sa druge, usmeri redefisnisanje uloge planera i pomogne im u traženju odgovora na operativna pitanja ko?, kako?, zašto?, šta?, koliko?, koja se svakodnevno postavljaju pred lokalnu zajednicu u konstekstu održivog urbanog razvoja. Rezultat istraživanja bio bi naučno zasnovan operativni model TIS-a za podršku održivom urbanom razvoju u Srbiji, struktrualno definisan na svim nivoima – sadržinskom, funkcionalnom i institucionalno - organizacionom, a metodologija njegove primene na lokalnom nivou potvrđena kroz pilot projekte. Naučni doprinos ovog istraživanja ogleda se u stvaranju novih osnova za dalje teorijsko i konceptualno redefinisanje sistema planiranja u Srbiji, preispitivanje uloge planera i planerskih agencija, i konačno osnov za razvoj novih metoda i tehnika planiranja u Srbiji obzirom na nov kvalitativno unapređen informacioni osnov.

The need for information is growing at all levels, from decision makers at the national and intrenational level to the lowest level of the individual. One of the big contemporary challenges is the development of information support to sustainable development. Efforts are focused on the development of ICT tools based on GIS technologies, which provide efficient collection, storage, updating, handling, analyzing and displaying all forms of data and information related to territory, necessary for the understanding of the territory in the process of collective decision making and the creation of sustainable policies and plans. In sustainable development, everybody is a user and information resource. Integrated approach to sustainability requires development and wide use of contemporary planners and management methods, tools, techniques and procedures that provide the best possible interpretation and integrated analysis of territory in group decision- making on specific actions and assessment of their impacts, risks and benefits for environmental, social and economic development. This research is motivated by the need to create conditions for effective participation of Serbia in the current global development process. At present, activities at local and regional level in the area of improving the information base for decision making and monitoring of integrated territorial development have focused on the development and application of the concept of territorial information systems based on ICT and GIS technologies in order to achieve greater efficiency administration, qualitative improvement of vertical and horizontal coordination, cooperation and rationalization processes in information and communication activities, with emphasis on raising the quality of the information base for decision making. The survey of international experience in developing and implementing this kind of models helps understanding the possibilities and limits of its application in Serbia. The subject of this research is development of territorial information system model to support sustainable urban development in Serbia, as a new instrument of communication and decision support tools in the process of collaborative planning and development management at the local level. The research is aimed at TIS model structuring that would be aplicable in the current conditions of change planning paradigms and practices of planning, but also in future when it is expected a new model of planning based on active participation of all actors of development to established. Application of the model TIS in Serbia should on the one hand, contribute to processes of change and improvements in the system of urban planning and spatial development, and on the other, direct role of planners redefinition to assist them in finding answers to operational questions of who?, As ?, why?, what?, how?, which are daily placed in front of the local community in terms of sustainable urban development. The result of this study should be scientifically-based operational model of TIS to support sustainable urban development in Serbia, structuraly defined at all levels - substantive, functional and institutional - organizational, and methodology of its implementation at the local level confirmed through pilot projects. The scientific contributions of this research is to create a new basis for further theoretical and conceptual redefinition of the planning system in Serbia, questioning the role planners and planners agencies, and ultimately the basis for the development of new methods and techniques of planning in Serbia as a new qualitatively improved information base.

Projektovanje seizmički otpornih arhitektonskih objekata korišćenjem panela od konstrukcionog stakla ili polikarbonata

Design of seismic resistant architectural structures by using panels of construction glass or polycarbonates

U tezi je predložen koncept projektovanja seizmički otpornih arhitektonskih objekata, upotrebom nekonvencionalnih materijala – lameliranog stakla i lameliranog klirita, koji se u građevinarstvu najčešće ne koriste u tu svrhu, pa su, iako široko rasprostranjeni i lako dostupni, definisani kao nekonvencionalni. Predloženim konceptom projektovanja, lamelirano staklo i lamelirani klirit, predviđeni su kao ispuna u čeličnom ramu, na koji naležu posredstvom intermedijalnog sloja gume, čime se formira kompozitni sklop. Na taj način se formiraju vertikalni elementi ukrućenja, integrisani u konstruktivni sistem objekta, osposobljeni za prijem seizmičke sile. Primenljivost takvog sistema, proverena je eksperimentom u laboratorijskim uslovima, gde su eksperimentalni modeli na bazi lameliranog stakla i lameliranog klirita izlagani dejstvu cikličnog opterećenja, koje simulira seizmičku silu. Time je proveren kapacitet nosivosti kompozitnih sklopova, za dejstvo dinamičkog opterećenja, paralelnog ravni sklopa. Utvrđen je intenzitet naprezanja kojem kompozitni sklopovi mogu biti izloženi, stepen ukrućenja konstrukcije u smislu iskazanih deformacija, kao i prednosti jedne vrste ispune u odnosu na drugu. Pomoću specijalizovanog softvera, čiji se rad bazira na metodi konačnih elemenata, kreiran je računarski model konstrukcije objekta koji je obrađivan u studiji slučaja, koji je u prvoj fazi procesa projektovanja služio za analizu problema. U taj računarski model, implementiran je sistem ukrućenja na bazi kompozitnih sklopova ispitanih u laboratoriji. Rezultati modalne analize i seizmičkog proračuna iz računarskog modela sa primenjenim ukrućenjima, pokazali su efikasnost rešenja, čime je zaokružena procedura projektovanja aseizmičkog ukrućenja korišćenjem nekonvencionalnih materijala.

The thesis proposes a design concept of seismic resistant architectural structures by using unconventional materials-laminated glass and laminated klirit, materials that are not usually used in construction design for that purpose, although being widespread and easily available, and therefore defined as unconventional. In the proposed design concept, laminated glass and laminated klirit are considered as a filling in a steel frame on which they overlap by the intermediate rubber layer, thereby forming a composite assembly. In this way vertical elements of stiffening are formed, capable for reception of seismic force and integrated into the structural system of the building. The applicability of such a system was verified by experiments in laboratory conditions where the experimental models based on laminated glass and laminated klirit had been exposed to the cyclic loads that simulate the seismic force. In this way the load capacity of composite assemblies was tested for the effects of dynamic load that was parallel to assembly plane. Thus, the stress intensity to which composite systems might be exposed was determined as well as the range of the structure stiffening referring to the expressed deformation along with the advantages of a particular type of filling compared to the other one. Using specialized software whose operation is based on the finite element method, a computer model of the structure was created and processed in the case study; the same computer model was used for analyzing the problem in the first phase of the design process. The stiffening system based on composite assemblies tested in laboratories is implemented in the computer model. The results of the modal analysis and seismic calculation from the computer model with stiffeners applied showed an efficacy of such a solution, thus rounding the design procedures for aseismic stiffening by using unconventional materials.

Analiza ektopičnog osteogenog potencijala mezenhimskih matičnih ćelija belog masnog tkiva kod miša na osnovu relativne ekspresije gena markera osteogeneze

Analysis of the ectopic osteogenic potential of adipose-derived mesenchymal stem cells in mice based on the relative gene expression of osteogenic markers

Osnovni cilj disertacije je da se uporedi i analizira osteogeni potencijal sveže izolovanih mezenhimskih matičnih ćelija belog masnog tkiva (ADSC) i in vitro osteoindukovanih ADSC na modelu ektopične implantacije kod Balb/c miševa. ADSC su nakon izolacije okarakterisane imunocitohemijskom analizom. Osteogena diferencijacija ADSC ispitana je analizom relativne ekspresije gena osteriksa (Sp7), osteokalcina (Bglap), alfa 1 lanca kolagena tipa I (Col1a1) i alkalne fosfataze (Alpl), vizuelizacijom fosfata i analizom ekspresije osteopontina. Miševima su subkutano implantirani implantati sastavljeni od sveže izolovanih ADSC, mineralnog koštanog matriksa (BMM) i plazme obogaćene trombocitima (PRP) (SPN grupa), osteoindukovanih ADSC, BMM i PRP (OPN grupa), PRP i BMM (PN kontrola) i samo od BMM (N kontrola). Osteogeneza u implantatima je ispitana analizom relativne ekspresije Sp7, Bglap, Col1a1 i Alpl, histomorfološkom, histomorfometrijskom i imunohistohemijskom analizom. Nađena je pozitivna CD29 i negativna CD14 ekspresija kod ćelija, što je karakteristika ADSC. Nakon petnaestodnevne osteoindukcije ADSC, zabeležena je najjača ekspresija markera osteogene diferencijacije. Najviša ekspresija gena markera osteogeneze, prisustvo ćelija nalik osteoblastima i osteoklastima, resorpcija BMM, visok procenat infiltriranog tkiva i prisustvo tkiva nalik osteoidu, ukazuju na napredovanje osteogeneze u OPN grupi nakon osme nedelje. U SPN grupi, nađeni su izraženi znaci inicijacije osteogeneze posle druge nedelje, a od četvrte ekspresija gena i ostalih markera opada što ukazuje na slabljenje osteogeneze. Rezultati doprinose razumevanju osteogenog potencijala različito pripremljenih ADSC što treba imati u vidu pri izboru načina i uslova za njihovu primenu u tretmanu koštanih defekata.

The main subject of this doctoral dissertation was to compare and evaluate the osteogenic potential of freshly isolated, non-treated adipose-derived mesenchymal stem cells (ADSCs) and the potential of in vitro osteoinduced ADSCs in the ectopic bone- forming model in Balb/c mice. The ADSCs were characterized after isolation by immunocytochemical analysis. Osteogenic differentiation of ADSCs was evaluated using relative gene expression analysis of osterix (Sp7), osteocalcin (Bglap), collagen type I alpha 1 chain (Col1a1) and alkaline phosphatase (Alpl), along with an assay for detection of phosphate deposits and an immunoexpression analysis of osteopontin. The implants composed of freshly isolated ADSCs, bone mineral matrix (BMM) and platelet-rich plasma (PRP) (SPN group), osteoinduced ADSCs, BMM and PRP (OPN group), BMM and PRP (PN control) and BMM-only (N control), were implanted subcutaneously in mice. The implants were analyzed using relative gene expression analysis of Sp7, Bglap, Col1a1 and Alpl, histochemical, histomorfometrical and immunohistochemical analyses. Positive immunoexpression of CD29 and negative of CD14 was found in the cells, which is a characteristic of ADSCs. After fifteen days of osteoinduction of ADSCs, the expression of the markers of osteogenic differentiation was the highest. The highest level of the bone-related gene expression, the presence of osteoblast-like and osteoclast-like cells, the resorption of BMM, a high percentage of infiltrated tissue and the presence of osteoid-like tissue, all suggest the progress of the osteogenesis in OPN implants after eight weeks. Strongly expressed signs of the initiation of osteogenesis were found in SPN implants after two weeks, but from the fourth week the expression of bone-related genes, as well as other analyzed markers decreased. The obtained results represent a contribution to the understanding of differences in the osteogenic potential of differently prepared ADSCs which is very important because of their utilization for the treatment of bone defects.

Analiza razlika u zaradama i višestrukog izbora zaposlenja primenom parametarskih i poluparametarskih ekonometrijskih metoda panela

An analysis of wage differences and multiple job holding by using parametric and semiparametric econometric methods for panel data

Predmet razmatranja u doktorskoj disertaciji jesu rodne razlike u zaradama i sklonost izbora više od jednog posla, kao prakse tržišta rada koje postaju naročito vidljive u prvim godinama tranzicije. Ove prakse su karakteristične za sve privredne sisteme, a ne samo za zemlje u tranziciji. Međutim, njihov obim i struktura variraju u zavisnosti od dinamike prilagođavanja faktora koji utiču na funkcionisanje tržišta rada. Stoga je praćenje ovih praksi važno, budući da mogu imati šire implikacije na socijalnu i ekonomsku politiku zemalja u tranziciji. Njihova analiza u doktorskoj disertaciji odnosi se na period intenziviranja procesa privatizacije u Srbiji sa početka 2000-ih godina i završava se periodom kada su počeli da se uočavaju prvi pozitivni efekti strukturnih reformi koji su zatim usporeni ekonomskom krizom iz 2008. godine. Testirane su dve osnovne istraživačke hipoteze: (i) da li postoje statistički značajne razlike u zaradama između muškaraca i žena i da li se te razlike jednim delom mogu objasniti uticajem diskriminacione prakse u određivanju zarada, (ii) kao i da li sklonost izbora više od jednog posla zavisi od zarade i časova rada na osnovnom poslu, očekivane zarade od dodatnog zaposlenja, te od ostalih determinanti koje utiču na odluku pojedinca da se istovremeno angažuje na više poslova. Obe hipoteze su empirijski potvrđene. Informacionu osnovu čine podaci Ankete o životnom standardu na bazi koje je formirana serija panel podataka za 2002. i 2003. godinu, kao i serija uporednih podataka za 2007. godinu. Polazeći od činjenice da se stope aktivnosti i zaposlenosti muškaraca i žena značajno razlikuju, rodni jaz u zaradama je posmatran uzimajući u obzir selekciju zaposlenih u sektor zarada. Rodni jaz je najpre izračunat na bazi modela izbora podataka za uporedne podatke ocenjenog metodom maksimalne verodostojnosti. Prema ovom metodu, ocenjeni rodni jaz se smanjio sa 10,96% na 5,97% tokom posmatranog perioda. U drugom koraku, rodni jaz je ocenjen na bazi poluparametarskog modela izbora podataka sa neparametarskom korekcijom pristrasnosti usled neslučajnog izbora pojedinca da radi za platu. Testirane su razlike u strukturi zarada ocenjene primenom parametarskog i poluparametarskog modela izbora podataka i zaključeno je da parametarska specifikacija odgovara modelu zarada. Treći metod koristi kovarijacioni metod ocenjivanja funkcije zarada u formi modela fiksnih efekata. Ovaj metod je potvrdio prisustvo statističke diskriminacije u određivanju zarada žena. Četvrti metod koristi kvantilne regresije u dekompoziciji rodnih razlika duž raspodele zarada. Primena ovog metoda dala je za rezultat veće rodne razlike na donjem levom delu raspodele (17,44%) a manje na desnom gornjem delu raspodele (9,12%), potvrđujući prisustvo efekta klizavog poda. Na kraju je ocenjen probit model sa slučajnim efektima, čije ocene su pokazale da je odluka da se neko opredeli za dodatni posao snažno finansijski motivisana, tj. da zarada od osnovnog posla smanjuje verovatnoću izbora za 20,8%, a očekivana zarada od dodatnog posla je poveća za 0,6%. Doprinos doktorske disertacije ogleda se u tome što se u njoj prvi put celovito analiziraju rodne razlike u zaradama u srednjoj vrednosti i na različitim kvantilima raspodele zarada svih zaposlenih u Srbiji tokom perioda intenzivnih strukturnih reformi. U analizi ovih praksi korišćeni su parametarski i poluparametarski modeli i ekonometrijske tehnike ocenjivanja za uporedne podatke i podatke panela. U disertaciji se razmatraju karakteristike zaposlenih koji pored osnovnog imaju i dodatno zaposlenje, i ocenjuju determinante koje utiču na verovatnoću istovremenog izbora više od jednog posla. Rezultati ove analize su dobijeni primenom nekoliko uporednih metodoloških pristupa i ukazuju na značajno prisustvo neobjašnjenog rodnog jaza u zaradama koji sadrži i skrivenu komponentu diskriminacije, kao i na značajnost faktora koji utiču na verovatnoću izbora dodatnog posla, pre svega, u neformalnom sektoru. Ključni nalazi analize sugerišu da fokus javnih politika treba usmeriti na one institucije koje mogu da osnaže pregovaračku poziciju žena i doprinesu suzbijanju neformalnih praksi na tržištu rada Srbije.

Doctoral dissertation considers gender wage differences and the propensity to hold multiple jobs as practices of the labour market that become especially visible during the first years of the transition to a market economy. These practices are characteristic of all the economic systems not only in transition economies. However, their scope and structure vary depending on the dynamics of adjustment of the factors influencing the functioning of the labour market. These practices may affect the social and economic policies of the transition countries and therefore monitoring them becomes important. Their analysis in this doctoral dissertation refers to the period of intensification of the privatization process of the early 2000s in Serbia and ends with a period when the first positive effects of structural reforms began to be perceived, which were then slowed down by the economic crisis of 2008. Two principal research hypotheses are examined: (i) are there statistically significant wage differences between men and women and can this difference be partly explained by an influence of discriminatory practice in wage determination; (ii) does the propensity to hold more than one job depend on primary job wages and hours of work, expected wages from a second job, as well as on other determinants that may affect individuals’ decisions to be engaged in more than one job simultaneously. Both hypotheses are empirically confirmed. The data are taken from the Living standards measurement survey, based on which a panel sample for 2002 and 2003, as well as a cross-section data set for 2007 were created. Since both the employment and participation rates of men and women differ substantially, the gender wage gap is examined taking into account the correction for sample selection bias of those who choose the employment in the wage sector. The gender wage gap is initially computed by using the maximum likelihood estimation of the sample selection model for cross-section data. According to this method, the gender wage gap has declined from 10.96 to 5.97 percent over the observed period. In a second step, the gender wage gap was estimated based on a semiparametric model of sample selection with nonparametric correction for potential bias due to non-random selection into the employment. The differences in the wage structure obtained by the parametric and semiparametric models of sample selection were tested and it is concluded that the parametric specification is the better fit for the wage function. The third method uses the within estimator of fixed effects model to estimate the wage functions. This method confirms the potential influence of statistical discrimination on the determination of women’s wages. The fourth method uses the quantile regression approach to decompose the gender wage gap along the wage distribution. This method yields a larger wage gap (17.44 percent) in the lower part and a smaller wage gap (9.12 percent) in the upper part of the wage distribution, indicating the existence of a sticky floor effect. The fifth method estimates the probit model with random effects and confirms the assumption that the decision to choose an additional job is strongly financially motivated, e.g. primary job wages reduce the probability of multiple job holding by 20.8 percent, while the second job reservation wage increases this probability by 0.6 percent. This doctoral dissertation contributes to the literature by providing a comprehensive analysis of the gender wage gap at both the mean and along the wage distributions of all employees in Serbia during the period of intensive structural reforms. In the analysis of these practices, parametric and semiparametric models are used together with the econometric methods of estimation of the cross-section and panel data. The dissertation examines the characteristics of those employees who have more than one job and the determinants that simultaneously affect the probability of multiple job holding. The main results of this analysis are obtained by using several comparative methodological approaches and they indicate the existence of a significant unexplained gender wage gap capturing wage discrimination and the significance of the factors affecting the probability of choosing more than one job, primarily in the informal sector. The findings suggest that the focus of the public policies should be directed towards those institutions that may strengthen the bargaining position of women and contribute to combating informal practices in the Serbian labour market.

Modeli integracije istorijskog centra i rečnog priobalja post-industrijskih gradova

Models of integration of the historical center and the river coastal area in post-industrial cities

Tema doktorske disertacije je istraživanje modela integracije istorijskog centra i rečnog priobalja post-industrijskih gradova, koja pripada području urbanističkog planiranja. Istorijski centar kao jedan od ključnih sastavnica gradskog prostora, je ,,telo“ koje se transformiše i prilagođava promenama rasta, što naposletku doprinosi urbanitetu i kvalitetu uslova života u gradu. Istraživanjem su utvrđeni principi oblikovanja istorijskog centra kao i urbanog područja rečnog priobalja, te je jasnije utvrđeno morfološko gledište na usmereni rast postojećeg istorijskog centra s ciljem integracije planiranog urbanog područja rečnog priobalja, u gradsku celinu. Za razumevanje istraživačkog problema, sagledane su specifične lokacije odabranih gradova (Beograd – Republika Srbija, Zagreb i Osijek – Republika Hrvatska), s istaknutom idejom njihove rekonstrukcije. Što se više povećava kompleksnost grada, u zavisnosti od njegove veličine, uočeno je da raste količina neplaniranog, spontanog i delimično usmerenog razvoja istorijskog centra. Zadatak istraživanja bio je dovesti delove rečnog priobalja u vezu s istorijskim centrom kao pojavom novog integrisanog kvalitetnog oblika korišćenja i namena površina. Nakon što je istraživanjem na izrađenim modelima proverena valjanost hipoteza, dokazano je na sva tri primera gradova da su istorijski centar i rečno priobalje najvažnije sastavnice suodnosa grada i reke, a njihova integrisanost ukazuje na vrednost celine grada. Integracija istorijskog centra i rečnog priobalja ostvariva je planiranjem usmerenog rasta istorijskog centra uz uvažavanje konteksta i identiteta grada. Planiranje prostorne integracije moguće je unutar ,,gravitacionog polja“ među razvojnim polovima, pri čemu su se pokazali različiti intenziteti uzajamnog privlačenja istorijskog centra i rečnog priobalja, (Osijek – intenzivno, Beograd – srednje intenzivno, Zagreb – nisko intenzivno). Na osnovu teza iskazani su scenariji za mere integracije istorijskog centra i rečnog priobalja kod daljeg urbanističkog planiranja. Realni scenario ostvariv je u Beogradu artikulisanjem usmerenog rasta istorijskog centra s varijacijama (dunavskim i savskim) uređenja rečnog priobalja i njihovom integracijom u prostorno fizički sklop. Pesimistički scenario integracije istorijskog centra i rečnog priobalja poželjno je promeniti u Zagrebu otklanjanjem konceptualnog procepa između planova uređenja ,,centralne ose“ i realizacije uređenja rečnog priobalja u cilju umanjenja njihove separacije. Optimistički scenario integracije istorijskog centra i rečnog priobalja u Osijeku oslanja se na kompleksnost i originalnost njihovog međuodnosa i izraza, dok je moguća transpozicija istorijskog centra u novo urbano područje rečnog priobalja na levoj obali reke. Činjenica da je neki grad nastajao pod određenim uticajem i u međusobnoj interakciji nepromenljiva je kategorija jer urbanističko planiranje sa artefaktima istorijskog centra i rečnog priobalja kao i njihova prošlost ne mogu se rekontekstualizovati. Rad je delom povest urbanizma Beograda, Zagreba i Osijeka iz razdoblja pripadnosti zajedničkoj državi.

The theme of the doctoral dissertation is a research of the model of the integration of the historical centre and the river coastal area in post - industrial cities, which belongs to the area of urban planning. The historical centre as one of the key components of urban space is the "body" that transforms and adapts to changes in growth. At the end, it contributes to the urbanization and quality of living conditions in the city. The research has determined the principles of the formation of the historical centre and the urban area of the river coastal area. Morphological point of view has been established on the directed growth of the existing historical centre with the aim to integrate the planned urban area of the river coastal area into the urban unit. In order to understand the research problem, specific locations of selected cities (Belgrade - Republic of Serbia, Zagreb and Osijek - Republic of Croatia) have been illustrated, together with an idea of their reconstruction. As the city complexity increases, depending on its size, the amount of unplanned, spontaneous and partially directed development of the historical centre is growing. The main task of the research was to bring the parts of the river coastal area in connection with the historical centre as a new integrated quality form of utilization and use of the surface. After checking the validity of the hypothesis, the three examples of cities have proved that the historical centre and the river coastal area are the most important constituents of the city and river, and their integrity points out the value of the city. The integration of the historical centre and the river coastal area can be realized by planning a directed growth of the historical centre, with respect to the context and identity of the city. Planning of spatial integration is possible within the "gravitational field" between the developmental poles. At that point, different intensity of mutual attraction of the historical centre and the river coastal area (Osijek - intensive, Belgrade - medium intensive, Zagreb - low intensive) has been determined. Based on the thesis, the scenarios for measuring the integration of the historical centre and the river coastal area are presented in the further urban planning. A realistic scenario can be accomplished in Belgrade by articulating a directed growth of a historical centre with variations (Danube and Sava) of the river coastal arrangements and their integration into a spatially physical frame. The pessimistic scenario of integrating the historical centre and the river coastal area should be changed in Zagreb by removing the conceptual gap between the plans of the "central axis" and the realization of the river coastal management in order to reduce their separation. The optimistic scenario of the integration of the historical centre and the river coastal area in Osijek relies on the complexity and originality of their interrelationship and expression. There is a possible transposition of the historical centre into the new urban area on the left bank of the river. The fact that one city has emerged under certain influence and interaction is an invariable category for urban planning. The artefacts of the historical centre and the river costal area, as well as their past, cannot be reconsidered. The dissertation is partly composed of the history of urbanism of Belgrade, Zagreb and Osijek from the period when they belonged to a common state.

Uticaj mezenhimskih ćelija belog masnog tkiva miša, indukovanih in vitro ka endotelskim i osteogenim ćelijama, na vaskularizovanost ektopičnih osteogenih implanata

Influence of adipose-derived mesenchymal stem cells in vitro induced into endothelial and osteogenic cells on vascularization of ectopic osteogenic implants

Koštano tkivo je dobro vaskularizovano, visokog kapaciteta regeneracije, ali su hirurške intervencije neophodne kod defekata kritične veličine. U tkivnom inženjerstvu kosti, problem nedovoljne vaskularizacije nema adekvatno rešenje, pa je s tim ciljem u ovoj disertaciji ispitivan uticaj mezenhimskih matičnih ćelija belog masnog tkiva (ADSC – Adipose-derived mesenchymal stem cells, engl.), indukovanih in vitro ka endotelskim (EĆ) i osteogenim ćelijama (OĆ), na vaskularizovanost ektopičnih osteogenih implanata. U ADSC dobijenim in vitro ekspanzijom ćelija stromalne vaskularne frakcije masnog tkiva BALB/c miševa muškog pola, pre i nakon ekspanzije je praćena imunoekspresija pozitivnih i negativnih ADSC-markera. ADSC iz trećeg pasaža su in vitro indukovane u EĆ ili OĆ ili ekspandirane bez induktivnih faktora. Tokom diferencijacije, praćena je ekspresija markerskih gena i proteina. Ekspandirane ADSC i plazma obogaćena trombocitima (PRP – Platelet-rich plasma, engl.) nanete na mineralni matriks kosti (MMK) su subkutano implantirane. Vaskularizacija i osteogeni proces u implantima (MMK, MMK/PRP, MMK/PRP/neindukovane ADSC, MMK/PRP/EĆ, MMK/PRP/OĆ, MMK/PRP/EĆ/OĆ) su procenjivani na osnovu ekspresije gena i proteina, histološke slike i histomorfometrijskih parametara. Ćelije iz trećeg pasaža su mezenhimske matične, a ADSC su diferentovane u EĆ ili OĆ. Implanti sa ili EĆ ili OĆ indukuju vaskularizaciju najpovoljniju za ektopični osteogeni proces. Dobijeni rezultati daju nove podatke o ćelijskim i molekularnim mehanizmima vaskulogenog i angiogenog procesa u osteogenezi i otvaraju nove mogućnosti u tkivnom inženjerstvu kosti i kliničkom tretmanu defekata koštano-zglobnog sistema.

Bone tissue is well vascularized, with high regenerative capacity, but surgical interventions are necessary in critical-sized defects. Since in bone tissue engineering (BTE), insufficient vascularization problem has no adequate solution, the influence of mesenchymal stem cells isolated from white adipose tissue (ADSCs – Adipose-derived mesenchymal stem cells) and in vitro induced into endothelial (ECs) and osteogenic (OCs) cells, on vascularization of ectopic osteogenic implants was examined in this dissertation. In ADSCs, obtained by in vitro expansion of stromal vascular fraction cells isolated from adipose tissue of male BALB/c mice, immunoexpression of positive and negative ADSCs-markers was examined before and after expansion. ADSC from the third passage were in vitro induced into ECs and OCs, or expanded without inductive factors. During differentiation, the expression of gene and protein markers was also examined. Expanded ADSCs and platelet-rich plasma (PRP) seeded onto bone mineral matrix (BMM) were subcutaneously implanted. Vascularization and osteogenic process in implants (BMM, BMM/PRP, BMM/PRP/uninduced ADSCs, BMM/PRP/ECs, BMM/PRP/OCs, MMK/PRP/ECs/OCs) were assessed on the basis of the gene and protein expression, histological picture and histomorphometrical parameters. The cells from the third passage were mesenchymal stem cells, and ADSCs were differentiated into ECs and OCs. Implants with ECs or OCs induced the most favorable vascularization for ectopic osteogenic process. The obtained results give some new data about cellular and molecular mechanisms of vasculogenic and angiogenic process during osteogenesis and open some new possibilities in BTE and in clinical treatments of skeletal and articular system defects.

Ekspresija i biohemijska i funkcionalna karakterizacija R-MC 17 antigena pacova

Expression and biochemical and functional characterization of R-MC 17 antigen in rat

Razvoj T limfocita u timusu je rezultat niza dvosmernih komunikacija između timocita i ćelija mikrosredine, posredovanih pretežno adhezivnim molekulima. U Institutu za medicinska istaživanja VMA je napravljen panel monoklonskih antitela (mAt) R-MC serije, koja prepoznaju timusne epitelne ćelije (TEC). Cilj ovog rada je bio biohemijska, imunohistohemijska i funkcionalna karakterizacija antigena koga detektuje R-MC 17 mAt. Ovo mAt detektuje antigen od 205 kDa u timusu pacova. Na osnovu poređenja biohemijskih karakteristika anti-DEC 205 mAt (HD83) i unakrsne imunoprecipitacije antigena iz lizata timusa pacova pomoću ova dva mAt zaključeno je da R-MC 17 mAt detektuje pacovski DEC-205 molekul. R-MC 17 antigen je jako ispoljen na kortikalnim TEC, medularnim timusnim dendritskim ćelijama (TDC), DC u parakorteksu i interfolikularnim zonama limfnog čvora i DC u periarteriolarnom limfnom omotaču slezine. Slaba imunoreaktivnost je detektovana na limfocitima u folikulima limfnog čvora i slezine i makrofagama u crvenoj pulpi slezine. Takođe, ekspresija DEC-205 molekula je detektovana na oko 40% ćelija TEC linije sa karakteristikama ćelija „dadilja“. R-MC 17 mAt podstiče vezivanje i emperipoezu timocita od strane ove epitelne linije. Istovremeno, ekspresija R-MC 17 antigena je podjednaka na subpopulacijama TDC pacova (CD11b+ i CD11b-), i ne menja se pod uticajem GM-CSF-a. Oba anti-DEC 205 mAt (R-MC 17 mAt i HD83 mAt) stimulišu apoptozu timocita u kokulturi sa TDC i ovaj proces je povezan sa povećanim vezivanjem timocita za TDC. R-MC 17 mAt inhibira proliferaciju singenih timocita i alogenih T limfocita pacova u kokulturi sa TDC u prisustvu anti-TCR (R7.3) mAt. Intratimusna aplikacija R-MC 17 mAt dovodi do smanjenja celularnosti timusa pacova što je povezano sa indukcijom apoptoze timocita. Fenotipska analiza subpopulacija timocita nakon intratimusne aplikacije R-MC 17 mAt pokazuje smanjenje ukupnog broja CD4+CD8+ i CD4+CD8-, a povećanje ukupnog broja CD4-CD8+ timocita, dok je ukupan broj zrelih CD5+TCRjako+ timocita nepromenjen. Sveukupno, ovi rezultati prvi put opisuju potencijalnu funkciju R-MC 17 antigena u timusu. Opisani mehanizmi mogu biti povezani sa procesima intratimusne diferencijacije timocita i prezentacije vi autoantigena u timusu od strane kortikalnih TEC i medularnih TDC, a što u krajnjem ishodu ima uticaj na procese selekcije (delecije autoreaktivnih klonova timocita).

Intrathymic development of T lymphocyte is mainly governed by an array of bi- directional interactions between thymocytes and thymic microenvironment, which are mostly provided by the adhesive molecules. The monoclonal antibody (mAb) panel of R-MC series, designed against thymic epithelial cells (TEC), was developed at the Institute of medical research (MMA). The aim of this doctoral thesis was focused on biochemical, immunohistochemical and functional characterization of the R-MC 17 antigen, detected by R-MC 17 mAb. R-MC 17 mAb recognizes the 205 kDa molecule expressed in the rat thymus. Based on similarity in biochemical features with an anti- DEC-205 mAb (HD83), and cross-immunoprecipitation of antigen from the thymic lysate with these two mAbs, R-MC 17 was considered to be the antibody specific for the rat DEC-205 molecule. Cortical TEC, medullary thymic dendritic cells (TDC), lymph node dendritic cells (DC) in paracortex and interfollicular zones, and spleen paraarteriolar-lymphoid sheaths DC, are highly positive with R-MC 17. Lymphocytes in lymph node and spleen follicles and red pulp macrophages in spleen, show weak R-MC 17 reactivity. About 40% of TEC-line cells with nursing characteristics express R-MC 17 antigen as well. R-MC 17 stimulates thymocyte binding and emperipoesis displayed by this cell line. Concomitantly, R-MC 17 antigen is equally expressed on two rat TDC subpopulations (CD11b+ and CD11b-), and cultivation with GM-CSF has no effect. R- MC 17, as well as HD83, stimulates thymocyte apoptosis in co-culture with TDC, and this process is associated with the increased binding of thymocytes to TDC. R-MC 17 mAb inhibits proliferation of syngeneic thymocytes and allogeneic T lymphocytes in anti-TCR (R7.3)-stimulated cocultures with TDC. Intrathymic injection of R-MC 17 mAb decreases thymic cellularity, which was related to the induction of thymocyte apoptosis. Phenotypic analysis of thymocyte populations after intrathymic application of R-MC 17 mAb, indicates decrease of the absolute number of CD4+CD8+ and CD4+CD8- , and increase of CD4-CD8+ thymocyte subpopulations, while the absolute number of mature CD5+TCRhigh thymocytes stays unchanged. Taken together, these results show, for the first time, a potential function of R-MC 17 antigen in rat thymus. Mechanisms viii described, are potentially involved in the processes of intrathymic differentiation and the presentation of autoantigen in the thymus by the cortical TEC and medullary TDC, having final implications in the processes of thymic selection (deletion of autoreactive thymocytes).

Istraživanje mogućnosti primene veštačke inteligencije u predviđanju performansi kočnog sistema motornih vozila

The potentional application of artificial intelligence in motor vehicles braking system performance

Osnovni zahtevi koji se postavljaju pred današnje kočne sisteme motornih i priključnih vozila, u pogledu bezbednosti vozila i saobraćaja, se odnose na njihovo dalje unapređenje kroz razvoj novih, inteligentnih, rešenja. Suština ovih zahteva jeste da se omogući pomoć vozaču kroz inteligentno upravljanje sistemima na vozilu, odnosno njihovim performansama u različitim, dinamički promenljivim, radnim uslovima. Pošto kočne performanse vozila zavise od performansi kočnica, koje funkcionišu na principima trenja i samim tim imaju vrlo nepredvidiv karakter, i od usklađenosti tih performansi sa trenutnim uslovima prijanjanja u kontaktu pneumatika sa tlom, koji se mogu intenzivno menjati tokom samo jednog ciklusa kočenja, realizacija ovih zahteva je izuzetno kompleksna. To je osnovni razlog za sprovođenje istraživanja u pogledu razvoja i implementacije inteligentnijih načina upravljanja performansama kočnog sistema na osnovu uslova prijanjanja u kontaktu pneumatik–tlo. U ovoj doktorskoj disertaciji su istraživane mogućnosti primene tehnika iz oblasti veštačke inteligencije u cilju modeliranja složenih dinamičkih uticaja radnih režima kočnica motornih vozila i uslova u kontaktu pneumatik–tlo, kao i predviđanja ovih uticaja u cilju upravljanja performansama kočnica, a time i performansama kočnog sistema, u toku ciklusa kočenja. Zbog nemogućnosti modeliranja složenih dinamičkih uticaja radnih režima kočnica motornih vozila na njihove izlazne performanse, odnosno na vrednosti klizanja u kontaktu pneumatika i puta pomoću klasičnih matematičkih metoda, uvedena je nova inteligentna metoda bazirana na dinamičkim veštačkim neuronskim mrežama i fazi logici. U skladu sa time, u ovoj disertaciji su istraživane mogućnosti primene dinamičkih veštačkih neuronskih mreža i fazi logike u cilju modeliranja, predviđanja i inteligentnog upravljanja performansama kočnica, odnosno performansama kočnog sistema. Predmetno istraživanje je usmereno ka razvoju sposobnosti kočnog sistema ka inteligentnom prilagođavanju sile kočenja dinamičkim promenama podužnog klizanja točka (pneumatika) u kontaktu sa putem u toku ciklusa kočenja. Ovakav koncept upravljanja performansama kočnog sistema, na osnovu prethodnih i trenutnih vrednosti posmatranih uticajnih veličina i identifikovanih uslova prijanjanja tokom kočenja, podrazumeva predviđanje potrebne vrednosti pritisaka aktiviranja kočnica, na prednjoj i zadnjoj osovini, za date uslove kočenja (vrednosti pritiska aktiviranja kočnice, vrednosti brzine točka na prednjoj/zadnjoj osovini, temperature u kontaktu frikcionog para kočnice na prednjoj/zadnjoj osovini i vrednosti klizanja u kontaktu pneumatik–tlo) kako bi se u kontaktu pneumatika i tla postiglo željeno (optimalno) klizanje u podužnom pravcu. Sve ovo je neophodno omogućiti uz razvoj mogućnosti inteligentnom kočnom sistemu da kontinualno uči, tokom svakog ciklusa kočenja, o složenim i stohastičkim uticajima između navedenih veličina. S obzirom da je ovo naročito važno za privredna vozila, fokus istraživanja je bio usmeren na mogućnosti unapređenja performansi elektronski upravljanog kočnog sistema, kako bi se u kontaktu pneumatika i tla ostvarila optimalna vrednost klizanja ne samo u podužnom pravcu, već i kasnija optimizacija klizanja u odnosu i na poprečno klizanje točka. Složena i dinamički promenljiva funkcionalna korelacija između pritiska aktiviranja kočnice i klizanja točka u podužnom pravcu tokom kočenja, brzine točka i temperature kočnice u toku ciklusa kočenja, modelirana je putem razvoja inverzne funkcionalne veze između ovih veličina. Razvojem inverznog dinamičkog modela procesa kočenja i njegovih efekata na podužno klizanje u kontaktu pneumatik-tlo, omogućeno je istovremeno učenje o toj funkcionalnoj vezi sa svakim novim kočenjem. Pored toga, omogućeno je i testiranje trenutnog „znanja“ dinamičkog modela, kao i podešavanje „performansi predviđanja“ modela novonastalim promenama podužnog klizanja točka. Ovakvim pristupom je omogućeno da željena dinamička promena klizanja u kontaktu pneumatika i tla može da bude postavljena na nivou koji obezbeđuje iskorišćenje maksimalnog prijanjanja točka (pneumatika) u podužnom i/ili bočnom pravcu. Ovim se obezbeđuju maksimalne kočne performanse, odnosno pruža mogućnost za optimizaciju sile kočenja u pogledu podužnog i bočnog usporenja, tj. stabilnosti vozila, posebno pri kočenju u krivini. Razvojem ovakvog dinamičkog modela, pomoću dinamičkih veštačkih neuronskih mreža, za kočnice prednje i zadnje osovine, stvoreni su uslovi za razvoj odgovarajućih neuronskih i neuro-fazi kontrolera za upravljanje, tj. selekciju pritisaka aktiviranja kočnica na prednjoj i zadnjoj osovini u zavisnosti od željenog klizanja, željenih kočnih performansi i potrebne raspodele sile kočenja. U skladu sa tim, u doktorskoj disertaciji su projektovani posebni neuro i neuro -fazi kontroleri u pogledu razvoja potrebnih „znanja“, tj. inteligentnih osobina kontrolera za dinamičko upravljanje pritiskom aktiviranja kočnica na prednjoj i zadnjoj osovini na osnovu željenog klizanja u kontaktu pneumatika i tla.

In terms of vehicle and traffic safety, the main demands imposed to the braking systems of motor vehicles and trailers are related to their further improvement through development of new, intelligent, solutions. It could enable the driver assistance function through an intelligent control of the vehicle systems performance in different and dynamically changing operating conditions. Since the braking performance of vehicles depend on the performance of the brakes, which based their function on the friction, it is a difficult to control stochastically changed the brakes performance. Furthermore, harmonization of that performance with the actual conditions in the tire-road contact, which is also intensively changed during a braking cycle, the realization of demands towards an intelligent control the braking system performance is very complex. This is the main reason for conducting research regarding development and implementation of more intelligent ways for control of the braking system performance. In this doctoral thesis, possibilities for employing of an artificial intelligence have been investigated in order to model and predict the impact of the brakes operating regimes and the complex conditions in the tire-road contact in order to provide intelligent controlling of the braking system performance during a braking cycle. Due to the impossibility for modeling of complex dynamic influences of brakes’ operating conditions on their performance and consequently on the value of the longitudinal wheel slip using conventional mathematical methods, a new method has been introduced based on an integration of dynamic neural networks and fuzzy logic. Accordingly, this thesis investigated possibilities for the proper integration of dynamic artificial neural networks and fuzzy logic in modeling, prediction, and intelligent control of the brakes’ performance, i.e. performance of the braking system. It should provide inherent capabilities of the braking system towards an intelligent adaptation of the braking forces to the dynamic changes of the longitudinal slip ratio in the tire–road contact during a braking cycle. This concept for control of the braking system performance, based on previous and current values of observed influential factors, means predicting of the brake applied pressure values, on the front and rear axle, for the given braking conditions (brake applied pressure, wheel speed on the front/rear axle, brake interface temperature on the front/rear axle, and wheel slip) in order to achieve the desired and/or optimal slip level in the longitudinal direction. Furthermore, the braking system should continuously learn about the complex and stochastic influences between these factors during a braking cycle. Since this is especially important for commercial vehicles, the focus of research has been directed on possibilities for improving the performance of electronically controlled braking system. It is done not only to achieve the optimal value of the longitudinal wheel slip in the tire-road contact, but also enables later optimization of the lateral wheel slip. The complex and dynamic changing a functional correlation between the brake actuation pressure and the wheel slip in the longitudinal direction, the wheel speed, and the brake interface temperature during a braking cycle is modeled through development of an inverse relationship between these quantities. The developed inverse dynamic model of the braking process, which includes the transfer of generated braking force from the tire to road, enabled the following: (a) the simultaneous learning about that functional relationship with each new brake application, (b) testing of the current "knowledge" of dynamic model, and (c) adjusting of the model’s prediction capabilities according to the new changes of the wheel slip. This approach enables setting of the desired dynamic change of the wheel slip in the contact of tires and road at the level that ensures utilization of the maximum wheel adhesion in the longitudinal and/or lateral direction, which in turn provides maximum braking performance and offers possibilities for the braking force optimization with respect to longitudinal and lateral deceleration. Developed the dynamic neural models of the braking process, for the front and rear axle, have created the conditions for the further development of appropriate neural and neural-fuzzy controllers that could be used for the control of braking system performance, i.e. selection of the brake actuation pressures depending on the desired wheel slip ratio, actual braking performance, and the brake force distribution. Accordingly, the separate neural and neuro-fuzzy controllers have been designed for dynamic control of the brake actuation pressures, on the front and rear axle, based on the desired wheel slip level in the contact between the tire and the road.

Tipologija i valorizacija građevinske stukture stambenih zgrada Beograda sa stanovišta komfora stanovanja

Typology and valorization of building structure of residential buildings of Belgrade from the standpoint of living comfort

Istraživanje karakteristika stambenih zgrada Beograda predstavlja preduslov za celovito sagledavanje stanja domaćeg stambenog fonda, njegovih kvaliteta ili nedostataka i osnovu za zaključivanje čime zaista raspolažemo i kakvi su dalji pravci delovanja. Predmet doktorske disertacije je istraživanje građevinske strukture stambenih objekata u Beogradu i njihovo vrednovanje sa stanovišta komfora u stanovanju. Ispitivanje je fokusirano na sve ono što u materijalnom smislu čini sklop objekta: konstrukcija, elementi omotača, elementi pregrađivanja i finalizacija i na njihov uticaj na ostvarivanje uslova komfora. Uzimajući u obzir da su vremenske odrednice u kojima je nastajao stambeni fond Beograda postavljene u rasponu od 150 godina, izdvojeni su karakteristični vremenski periodi koji predstavljaju reprezentativne celine građevinskih dostignuća. Ovakav pristup imao je za cilj struktuiranje obimne stručne građe i tipološko izdvajanje onoga što čini reprezentativni uzorak. U okviru posmatranog perioda identifikovani su tipični sklopovi i izvršena je sistematizacija, uočene su karakteristike u građenju i istražena građevinska regulativa, što je poslužilo kao osnova za formiranje strukture reprezentativnih modela, na kojima je sprovedena analiza kvaliteta stambenog komfora koji podrazumevaju: toplotni, akustički, vazdušni i svetlosni komfor. Sledeći korak u istraživanju predstavljalo je definisanje parametara svakog pojedinačnog komfora i formiranje metodologije kojom će se vršiti ispitivanje ustanovljenih modela stambene izgradnje. S obzirom da se svakim pojedinačnim komforom bave posebne specijalizacije, ili čak posebne struke, kompleksnost materije koja je trebala biti analizirana utoliko je bila složenija, što je zahtevalo različite metodološke postupke i multidisciplinarnost u istraživačkom radu. Parametri su odabrani tako da svaka kategorija zahteva daje izmerljive rezultate, čime je izbegnuta nargumentovana procena, i sprovedeno objektivno vrednovanje. Uz pomoć računarskih alata koji su specijalizovani za pojedinačne kategorije komfora (termički, zvučni i svetlosni) istraženi su reprezentativni modeli i dobijeni rezultati na osnovu kojih su izvedeni zaključci o kvalitetu stambenih zgrada Beograda. Rezultati su prvo struktuirani po kategorijama komfora, a zatim su sumirani komfori po pojedinačnim modelima, kako bi se objedinile karakteristike svakog pojedinačnog uzorka, izvela sintezna valorizacija, i formulisali zaključci o ustanovljenim hronološkim celinama.

The survey of characteristics of residential buildings of Belgrade represents a pre-requisite for a comprehensive assessment of the state of the domestic housing stock, its qualities or flaws, and the basis for the conclusion of what we actually have and what are the further courses of action. The subject of this dissertation is to explore the building structures of residential buildings of Belgrade and their further evaluation from the standpoint of comfort or residence. The study was focused on everything that in a material sense makes a building structure (construction, elements of building envelope, partition elements and finalization) and their impact on achieving requirements of comfort. Taking into account that the timelines in which the Belgrade housing stock was produced is set in the range of 150 years, typical time periods were established in relation to building achievements relevant for the given time. This approach was aimed at structuring extensive professional material and typological positioning of what constitutes a representative sample. Within the observed period, typical structures were identified and systematized, characteristics in building were observed and building regulations were investigated, which served as the basis for the formation of representative models and their structure. On such established models, analysis of quality of living comfort was conducted, including: thermal, acoustic, and visual comfort, as well as indoor air quality. The next step in the research is the definition of relevant parameters of each individual comfort and the formation of the methodology that will be used for testing of established models of housing construction. The fact that every aspect of the comfort is subject of a unique specialization or profession caused different methodological procedures and a multidisciplinary research. The parameters are chosen so that for each category of requirements measurable results are obtained, enabling an objective evaluation. With the help of computer tools which are specialized for particular categories of the comfort (thermal, sound and light), representative models are explored and results are obtained, on the basis of which the conclusions about the quality of residential building in Belgrade are drawn. Results are structured by categories of comfort and then summarized by individual models, in order to consider the characteristics of each sample comprehensively, to perform synthesized valorisation, and formulate conclusions on the established chronological unities.

Morfološka varijabilnost kinetičkog glavenog skeleta i pileusa zmija iz roda Natrix (N. natrix i N. tessellata)

Morphological variation of kinetic head skeleton and pileus in snake from the genus Natrix (N. natrix and N. tessellata)

Većina dosadašnjih istraživanja strukture i obrazaca varijabilnosti glavenog skeleta odnosila su se na grupe sa akinetičkom lobanjom. Istraživanje kinetizma u okviru glavenog skeleta zmija može doprineti boljem razumevanju odnosa razvojne i funkcijske integracije, uticaja alometrije na varijabilnost oblika i morfološku integraciju glavenog skeleta, promena oblika tokom ontogenije, kao i morfološke divergencije između polova. Pored toga, analizama varijabilnosti pileusa može se steći uvid u opštu sliku varijabilnosti glavenog regiona. Osnovni zadatak ove disertacije je utvrđivanje obrazaca morfološke varijabilnosti i faktora koji utiču na varijabilnost glavenog skeleta i pileusa kod dve sestrinske vrste zmija iz roda Natrix (belouške, N. natrix i ribarice, N. tessellata) primenom metoda kompjuterizovane tomografije i geometrijske morfometrije. Rezultati su pokazali da se belouška i ribarica razlikuju u veličini i obliku elemenata glavenog skeleta. Uočene razlike najverovatnije odslikavaju razlike u tipu i veličini plena, sredini u kojoj love, strategijama lova i sposobnostima držanja i gutanja plena. Polni dimorfizam u veličini i obliku elemenata glavenog skeleta veoma je izražen kod belouške i ribarice. Kod obe vrste moždana čaura i elementi trofičkog aparata su visoko integrisani, dok se kosti njuške ponašaju kao zasebni moduli. Funkcija najviše doprinosi integrisanosti elemenata glavenog skeleta. Utvrđeno je da se tek rođene belouške i ribarice razlikuju u obliku pileusa, dok se unutar vrsta polni dimorfizam u obliku pileusa ispoljava tek kod adulata. Između moždane čaure i pileusa ustanovljeni su visoki nivoi korelisanosti, kako za veličinu, tako i za oblik.

Most of the previous investigations on the structure and patterns of cranial skeletons variability were done in groups with akinetic skull. Research of snake’s kinetic cranial skeleton can provide a better understanding of the relationship between developmental and functional integration, influence of allometry on shape variation and morphological integration of the cranial skeleton, shape changes during ontogeny, as well as morphological divergence between the sexes. In addition, analysis of pileus variability may provide insight into general pattern of head variability. The main aim of this thesis was to define the patterns of morphological variability and factors that affect the variability of head skeleton and pileus in two sister snakes species from the genus Natrix (Grass snake, N. natrix and Dice snake, N. tessellata) using the methods of computed tomography and geometric morphometrics. The results showed that N. natrix and N. tessellata differ in size and shape of cranial elements. Observed differences probably reflect differences in the type and size of the prey, hunting environment and strategies, handling abilities and prey swallowing. Sexual dimorphism in size and shape of cranial elements are pronounced among Grass and Dice snakes. In both species, braincase and elements of the trophic apparatus are highly integrated, while the bones of the snout can be considered as separate modules. The function is the main factor of morphological integration between cranial elements. Hatchlings of Grass and Dice snakes differed in pileus shape, while sexual dimorphism in pileus shape was detected in adults only. For both size and shape, high levels of correlation between braincase and pileus were estimated.

Linearno i nelinearno modeliranje geodetski registrovanih deformacionih procesa konstrukcija

Linear and nonlinear modeling geodetic registred deformation processes of structures

Predmet ovog rada je modeliranje ponašanja konstrukcija i okolnog tla na osnovu postupaka identifikacije sistema, odnosno eksperimentalnih podataka. U ovom radu su objašnjene sve procedure identifikacije sistema i njihova primena u deformacionoj analizi konstrukcija. Pozanto je da se ponašanje konstrukcije i tla modeluje na osnovu geomehaničkih i statičkih proračuna. Geomahanički proračuni se baziraju na osnovu uzetih uzoraka tla. Ovi uzorci ne mogu da reprezentuju ponašanje celog tla na kome se gradi objekat. Statički proračun se bazira na pravilničkim odredbama. Međutim, prilikom izgradnje objekta dolazi do grešaka (kvalitet ugrađenog materijala i propusta prilikom gradnje) i neobezbeđivanja projektovane geometrije objekta. Svrha modeliranja na osnovu eksperimentalnih podataka je: provera proračuna, sprečavanje ljudskih i materijalnih katastrofa kao i poboljšanje standarda izgradnje. Poznato je da se monitoringom konstrukcija i tla bave: građevinska, geološka i geodetska struka. Sve ove struke moraju da sarađuju i koriste istu terminologiju da bi se uspešno izvršio monitoring konstrukcija. Rezime Geodetska metodama meri se samo odgovor konstrukcije (izlazne signale) i jedina određuje pomeranja i deformacije u apsolutnom sistemu. Praktično istraživanje je podrazumevalo: analizu podataka vremenskih serija geodetskog monitoringa u vremenskom i frekventnom domenu i mogućnosti korišćenja ovih podataka za definisanje reda sistema i stepena modela, sagledavanje kompleksnosti eksperimentalnog modeliranja deformacionih procesa primenom postupak identifikacije sistema, mogućnost aproksimacije nelinearnih dinamičkih procesa konstrukcije linearnim, predlog postupka određivanja vremenskog plana opažanja na osnovu eksperimantalno određenog modela (identifikacija sistema). Ovo istraživanje je izvrešeno na simuliranim podacima vremenskih serija sleganja konstrukcije i merenim podacima pomeranja pilona mosta "Sloboda" u Novom Sadu prilikom ispitivanja konstrukcije na probno opterećenje. Prvo je analizirana značajnost obezbeđivanja stacionarnosti ili slabe stacionarnosti procesa. Stacionarnost se obezbeđuje postupkom detrendovanja podataka, odnosno uklanjanjem trenda procesa. Ovo je veoma zametan posao i pokazano je da nije neophodno obezbediti stacionarnost procesa. Dobijene razlike u oceni parametara modela za stacionaran i nestacionaran proces su zanemarljive. Za analizu podataka u vremenskom domenu korišćeni su koeficijenti autokorelacije, parcijalne autokorelacije i međusobne korelacije, a u frekventnom domenu Šusterov periodogram. Pokazano je da se na osnovu grafičke predstave podataka može definisati red sistema, koji je za konstrukcije prvog (sleganje i uticaj temperature) ili drugog reda (opterećenje konstrukcije). Koeficijent korelacije daje dragocene informacije o stepenu modela, ali se za dobijanje definitivnog modela mora testirati značajnost parametara. Za linearno modeliranje primenjeni su modeli funkcije prenosa (regresioni modeli) i modeli prostora stanja – metoda potprostora (Ojlerova metod i Tustinova aproksimacija), a za nelinearno veštačka neuronska mreža (mreža širenja unazad) i regresioni model za nelinearne procese. Takođe, primenjen je za modeliranje sleganja konstrukcije tzv. najčešće korišćene funkcije prenosa Rezime koje se koriste u upravljanju sistemima. Ocenjivanje parametara modela je izvršeno metodom najmanjih kvadrata, nelinearnim metodom najmanjih kvadrata i metodom greške predikcije. Na osnovu dobijenih kriterijuma aproksimacije za simulaciju i predikciju zaključeno je da je autoregresivni model za spoljašnjim ulaznim signalima, za slučaj linearnog ili nelinearnog procesa, najpodesniji za modeliranje ponašanja konstrukcija. Model potprostora stanja (Ojlerova metoda) se može koristiti za predikciju jedan korak unapred, a pretpostavljeni oblik funkcije prenosa (najčešće korišćene funkcije prenosa) je veoma pogodan za modeliranje sleganja konstrukcije (prednost je što je potreban mali broj epoha merenja za određivanje parametara modela). Model mreže širenja unazad (veštačka neuronska mreža) je adekvatan za simulaciju ponašanja konstrukcije. Procenat aproksimacije bio bi monogo bolji da je broj epoha merenja veći, što je inače retko u geodetskoj deformacionoj analizi. Analiza perioda odabiranja je pokazala da se epohe merenja treba da definišu po teoremi o odabiranju, koja određuje period odabiranja na osnovu prelaznog stanja sistema. Prelazno stanje sistema se određuje na osnovu projektovanog ponašanja konstrukcije na početku izgradnje, a posle na osnovu ekpsrimentalnih podataka.

The subject of this dissertation is modeling the behavior of structures and surrounding ground using system identification procedures, respectively based on experimental data. This paper explains all the procedures of identification systems and their application in deformation analysis of structures. It is known that structural and surrounding ground behavior is modeled on the basis of geomehanical and static calculations. Geomehanical calculations are based on samples taken on the ground. These samples may not represent the behavior of the whole ground on which build an structure. Statically analysis is based on rule book regulations. However, during construction of the building leads to errors (quality materials and omission during the building) and failure to ensure the projected geometry. The purpose of modeling based on experimental data is checking the project, preventing human and material disasters and improve the standard of construction. It is known that the monitoring of structures and ground deal in: civil engineering, geology, and geodetic professions. All these professions must work together and use the same terminology in order to successfully performed construction monitoring. By geodetic Abstract method we measures only the structural responses (output signals) and it is only profession which determines the movement and deformation in the absolute system. Practical research is meant: analysis of time series of geodetic monitoring in time and frequency domain and possibility of using these data to define the system order and order of models, analyzing the complexity of experimental modeling of deformation processes using the system identification procedure, the possibility of approximation of nonlinear dynamical processes by linear models, the proposed procedure determining the sampling of observations based on experimentally determining model (system identification). This research was derived on the simulated time series of settlement construction and the measured data of displacements of the pylon of the bridge, "Sloboda" in Novi Sad during test load of structure. The first analyzes the importance of ensuring stationary or weak stationary of the process. Stationary of the process provides by detrending data, trend removed from process. This is a very hardly job and it was shown that it is not necessary to ensure stationary of the process. The differences in the estimated parameters of model for the stationary and no stationary process are negligible. For data analysis in time domain used coefficient of autocorrelation, partial autocorrelation and cross correlation, in the frequency domain used Schuster periodogram. It has been shown that graphical representations of data can be used to defined order of system, which can be for the structure first (subsidence and temperature effects) or second order (the loads). The correlation coefficient provides valuable information about the order of the model, but for a definite model must test the significance of parameters. For linear modeling were applied transfer function models (regression models) and state space models – subspace method (Euler method and Tustin approximation), for a non-linear artificial neural network (backpropagation neural network) and the pseudolinear regression model for nonlinear processes. Also, it was used so-called commonly used transfer functions, which are used in management systems, for modeling the settlement of construction. Parameters of the model are estimated using least squares, nonlinear least squares method and the prediction error method. On the basis of the criteria approximation for simulation and prediction, it was concluded that the autoregressive model with external input signals, in the case of linear or nonlinear Abstract process, the most suitable for modeling the behavior of structures. Subspace method (Euler method) can be used to predict one step ahead, a supоsed form of transfer function (the most commonly used transfer function) is very suitable for modeling settling of the structure (the advantage is that it requires a small number of epochs of measurement for determining the model parameters). Backpropagation neural network (artificial neural network) is adequate for simulation of the structural behavior. Percentage of approximation would be better that there are many data in time epoch, which is rare in the geodetic deformation analysis. Analysis of sampling period showed that the epoch of measurement should be defined by the Sampling theorem, which determines the sampling period based on the transition state of the system. Transition state of the system is determined on the basis of projected structural behavior at the start of building, and after based experimental data.

Tip stanovanja i kvalitet života osoba sa intelektualnom ometenošću

The housing type and life quality of people with intelectual disability

Stanovanje uz podr š ku za osobe sa intelektualnom ometeno šć u (IO) predstavlja uslugu u sistemu socijalne zašt ite koj a im omogu ć ava š to samostalniji ž ivot u najmanje restriktivnom okruženju . Primenom prog r ama usluge stanovanja uz podršku obezbeđuje se odgovarajući smeštaj, stručna pomoć i podrška za što potpunije osamostaljivanje i uključivanje korisnika u društvenu zaje dnicu . U cilju pobolj š anja kvaliteta usluge stanovanja uz podršku neophodno je uzeti u obzir objektivni kvalitet rezultata primene programa i subjektivni do ž ivljaj korisnika usluga, a iskustvene podatke potkerepiti istraživanjima u pravcu povezanosti između tipa stanovanja i kvaliteta života osoba sa IO i faktora koji njega utiču. Ciljevi istraživanja su utvrđivanje prirode odnosa kvaliteta života i tipa stanovanja osoba sa IO ; intenziteta i pojavnih oblika problematičnog ponašanja osoba sa IO u različitim tipovima sta novanja i nivoa doživljenog stresa u odnosu na tip stanovanja osoba sa IO. Hipoteze istraživanja su:  Osobe sa IO koje su obuhvaćene programom stanovanja uz podršku imaju bolji kvalitet života u odnosu na ispitanike koji žive u porodičnom okruženju i u stacionarn im ustanovama.  Intenzitet i pojavni oblici problema u ponašanju osoba sa IO zavise od tipa stanovanja.  Ispitanici koji su obuhvaćeni programom stanovanja uz podršku imaju niži nivo stresa u odnosu na ispitanike koji borave u instituciji ili u primarnoj porod ici. Istraživanjem su u okviru ispitivanog uzorka obuhvaćene odrasle osobe sa umerenom IO, starosti od: 17 - 79 godina. Prosečna starosna vrednost na nivou celokupnog uzorka 39,95 godine. Ukupan broj ispitanika je 122. Ispitivani uzorak je podeljen u tri grup e: osobe sa IO koje su smeštene u ustanovama stacionarnog tipa ( n = 51) ; osobe sa IO koje žive u svojim primarnim porodicama (n= 38) i osobe sa IO koje su obuhvaćen e p rogramom stanovanja uz podršku (n= 33). Prema polnoj strukturi i po pitanju zastupljenosti medic inskih smetnji bez obzira na tip stanovanja ispitanika ne postoji statistički značajna razlika. Instrumenti koji su korišćeni u istraživanju su: Sveobuhvatna skala za procenu kvaliteta života ( Comprehensive Quality Of Life Scale; ComQol – I5 , Cummins); Skal a neprijateljstva i agresije za odrasle, Reaktivna/Proaktivna ( The Adult Scale of Hostility and Aggression, Reactive/Proactive A - SHARP , Matlock, Aman); Upitnik o f u nkcijama ponašanja ( Questionnaire About Behavioral Function, QABF, Valmer , Maison) i Upitnik z a procenu stresnih životnih događaja ( Life Stress Inventory , Fogarty , Bramston , Cummins ). Rezultati našeg istraživanja pokazuju da o sobe sa IO , koje su obuhvaćene programom stanovanja uz podršku izražavaju veće zadovoljstv o kvalitetom života u odnosu na osta l a dv a poduzorka ( p=0,000) i imaju bolji kvalitet života u odnosu na ispitanike koji žive u instituciji . Prema rezultatima istraživanja problemi u ponašanju osoba sa IO, i ntenzitet i oblici ispoljavanja agresivnog ponašanja i nivo doživljenog stresa povezani su s a tipom stanovanja . Problemi u ponašanju statistički značajno su naj izraženiji kod ispitanika koji stanuju u instituciji, a najmanje izraženi kod ispitanika koji su u programu stanovanja uz podršku ( p=0,000) . Agresivno ponašanje je najmanj e izražen o kod ispitan ika sa IO koji su obuhvaćeni programom stanovanja uz podršku ( verbalna agresija (p=0,001), buling (p=0,002), prikrivena agresija (p=0,003), izražavanje neprijateljstva (p= 0,002) i fizička agresija (p= 0,001) ) . Rezultati ukupnog skora dobijenog na Upitnik u za p rocenu stresnih životnih događaja pokazuju da osobe koje su obuhvaćene programom stanovanja uz podršku u odnosu na ispitanike koji stanuju sa porodicama i u instituciji najmanje ispoljavaju s tres ( p=0,000) . R ezultat i ukazuju n a to da je tip stanovanja od veli kog značaja za sveukup an kvalitet života i opšte psihosocijalno stanje osoba sa IO . Ovo istraživanje može da da doprinos unapređenju kvaliteta implementacije programa stanovanja uz podršku za osobe sa IO i uključivanj u još većeg broja korisnika u taj program.

Supported housing for the people with intelectual disabilities (ID) represents the favour in the system of social protection which enables them more independen t life in the least restrictive surrounding. Adequate accommodation, skilled help and support for more complete independance and participation of the users in the social comunity are provided by applying the favour of programme of supported housing.In the aim of improving the quality of the favour of supported housing it is neccessary to take into consideration the objective quality of the results of the programme applying and subjective experience of the favour users, and experienced data should be confi rmed by researches in the direction of the connection between the type of life arrangement and the quality of life for people with ID and factors that have some influences to them. The aims of the research are comfirming the nature of the relationship in the life quality and type of housing for the people with ID; the intensity and appearing ways of troubled behaviour of people with ID in different types of housing and the level of experienced stress according to the type of housing for the people with ID. Hypothesis of the research are: 1. The people with ID that are overcome by the supported housing have better quality of life in the relation to the tested people who live in the family surrounding and in the stationary institutions. 2. The intensity and appea ring ways of troubled behaviour of the people with ID depend on the type of housing. 3. the tested people who are overcome by the programme of supported housing have lower level of stress in the relation to the tested people that reside in the institution or in the primary family. By the research are overcome, in the frame of the tested samples, adult people with moderate ID, age from 17 to 79 years, average age value on the level of the whole sample is 39,95 years. The total number of tested people is 122. Th e tested sample is devided into three groups: the people with ID that are situated in the institutions of the stationary type (n= 51) ; the people with ID that live in their primary families (n= 38) and the people with ID that are overcome by the programme of supported housing (n= 33). According to the sex structure and according to the presence of medical difficulties, no matter to the type of housing of tested people, there are no statistic important differences. The instruments that were used in the res earch were: Comprehensive Quality Of Life Scale ( Comprehensive Quality Of Life Scale; ComQol – I5 , Cummins); The Adult Scale of Hostility and Aggression, Reactive/Proactive ( The Adult Scale of Hostility and Aggression, Reactive/Proactive A - SHARP , Matlock, Aman); Questionnaire About Behavioral Function ( Questionnaire About Behavioral Function, QABF, Valmer , Maison) and Life Stress Inventory ( Life Stress Inventory , Fogarty , Bramston , Cummins ). The results of our research show that people with ID, who are ove rcome by the programme of supported housing express more satisfaction by the quality of life in the relation to the other two sub - samples ( p=0,000) and have better quality of life in the relation to the tested people who live in the institution. According to the results of the research, the troubled behaviour of people with ID, the intensity and ways of expressing aggressive behaviour and level of experienced stress are connected with the type of housing. The troubled behaviour are the most expressed, whic h is statistic important, at tested people who live in the institution and least expressed at people who are in the programme of supported housing ( p=0,000) . Aggressive behaviour is least expressed at tested people with ID who are overcome by the programme of supported housing ( verbal aggression (p=0,001), bulling (p=0,002), hidden aggression (p=0,003), expressing the hostility (p= 0,002) and physical aggression (p= 0,001) ) . Results of the total amount that was got on Life Stress Inventory show that the peple who are overcome by the programme of supported housing in the relation to the tested people who live with families and in the institutions express stress least ( p=0,000) . The results show on that the type of housing has the great importance for the whole quality of life and general psycho - social condition of the people with ID. This research can give the contribution to the improving of the implementation quality of programme of supported housing for the people with ID, including even bigger number of use rs in those programmes.

Studija asocijacije varijanti u genima za mikroRNK i za proteine utišavajućeg kompleksa sa rizikom za razvoj i progresiju karcinoma prostate kod bolesnika iz Srbije

The study of association of variants in genes encording microRNAs and the proteins of silencing complex with prostate cancer risk among patients from Serbia

Karcinom prostate (KP) predstavlja jedan od vodećih globalnih zdravstvenih problema, s obzirom na to da je reč o drugom najučestalijem malignom tumoru kod muškaraca. U kliničkoj praksi vezanoj za KP jedan od najvećih izazova je razlikovanje latentnog od klinički značajnog karcinoma. U tom smislu, od izuzetnog značaja je identifikacija genetičkih varijanti čija bi primena kao nestandardnih prognostičkih parametara mogla da se koristi za konstruisanje preciznih algoritama za procenu rizika za razvoj i progresiju KP. U okviru ove disertacije analizirana je potencijalna asocijacija varijanti u četiri gena za mikroRNK (rs2910164 u hsa-miR-146a, rs3746444 u hsa-miR-499, rs11614913 u hsa-miR-196a2 i rs895819 u genu za miR-27a) sa rizikom za razvoj i progresiju KP. Takođe, analizirana je asocijacija varijanti u genima za proteine utišavajućeg kompleksa (RISC) (DICER1, AGO2, TARBP2, GEMIN3 i GEMIN4) sa navedenim rizicima. Studija je obuhvatala 355 bolesnika sa KP, 360 bolesnika sa benignom hiperplazijom prostate (BPH) i 318 muškaraca koji su činili kontrolnu grupu. Genotipizacija genetičkih varijanti vršena je metodama PCR-RFLP, HRMA, alel- specifičnog PCR-a, primenom TaqMan® eseja za genotipizaciju polimorfizama i PCR-a u realnom vremenu baziranom na korišćenju specifičnih proba. Rezultati genotipizacije su statistički obrađeni u cilju ispitivanja asocijacija sa rizikom za razvoj i progresiju KP, kao i statističkih epistatičkih interakcija. Takođe, sprovedena je meta-analiza koja je uključivala naše rezultate, kao i rezultate prethodnih studija koje su analizirale asocijaciju rs2910164 sa rizikom za razvoj drugih malignih tumora. Rezultati ove studije pokazali su asocijaciju rs2910164 sa rizikom za razvoj bolesti, dok je za gotovo sve ostale analizirane genetičke varijante ustanovljena asocijacija sa vrednostima standardnih prognostičkih parametara KP i/ili rizikom za progresiju KP. Pokazana je i asocijacija epistatičke interakcije između varijanti rs11614913 u genu hsa-miR-196a2, rs7813 u genu GEMIN4, kao i rs784567 u genu TARBP2 sa rizikom za progresiju bolesti. Za rs2910164 u meta-analizu ustanovljena je asocijacija sa rizikom za razvoj više tipova malignih tumora.

Being the second most common cancer among men worldwide, prostate cancer (PCa) is one of the major global health issues. In the clinical practice related to this malignant disease, one of the most important challenges is to distinct latent from clinically significant forms of disease. Therefore, it is of utmost importance to identify genetic variants which could be used as nonstandard prognostic parameters for constructing precise algorithms for evaluating the risk of PCa onset and progression. In this dissertation, the association of four microRNA genetic variants (rs2910164 in hsa-miR- 146a, rs3746444 in hsa-miR-499, rs11614913 in hsa-miR-196a2 and rs895819 in the gene encoding miR-27a) with PCa risk and progression was assessed. Also, the potential association with PCa was estimated for genetic variants located in genes encoding the proteins of RNA-induced silencing complex (RISC) (DICER1, AGO2, TARBP2, GEMIN3 and GEMIN4). The study involved 355 patients with PCa, 360 patients with benign prostatic hyperplasia (BPH), as well as 318 healthy controls. Genotyping was performed by using PCR-RFLP, HRMA, alelle-specific PCR, TaqMan® SNP Genotyping Assays and real-time PCR-based genotyping assay using specific probes. Results were statistically analyzed, aiming to evaluate the potential associations of genetic variants with PCa risk and progression, as well as to assess statistical epistatic interactions. Furthermore, a meta-analysis was conducted involving our results, together with the results of previous studies on association between rs2910164 and different types malignant tumors. The results of this study suggested the association of rs2910164 with PCa risk, while for most of the other analyzed genetic variants the association with the values of standard prognostic parameters and/or PCa progression was found. Also, the epistatic interaction between rs11614913 in hsa-miR- 196a2, rs7813 in GEMIN4 and rs784567 in TARBP2 was shown to be associated with the risk of PCa progression. For rs2910164, the meta-analysis showed the association with multiple types of malignant tumors.

Istočnohrišćanska trijadologija u arhitekturi vladarskih zadužbina srednjovekovne Srbije

The principle of triadology in architecture of the Serbian mediaveal rulers' endowments

Srpska srednjovekovna arhitektura — arhitektura vladarskih zadužbina, bila je i ostala materijalizovano svedočanstvo recepcije vizantijske, a time i istočnohrišćanske kulturne tradicije, sa kojom su nasleđena prožimanja, u biti antinomičnih, tradicija antičke i judaističke misli — što je iskazano kroz dogmu o Svetoj trojici. Odgonetanje načela trojičnosti čini polaznu postavku za prepoznavanje teorijskog modela kojim je moguće tumačiti sakralnu arhitekturu srednjovekovnog perioda. Razvoj hrišćanskog dogmata i rešavanje pitanja o Bogu kao Trojici bilo je obeleženo zavisnošću hrišćanske misli od najznačajnijih judaističkih i poznoantičkih ideja, kao i filozofskih pojmovnih konstrukcija. Analiziranjem i praćenjem njihovog usvajanja uočava se napredovanje u rešavanju pitanja poimanja Jednog i mnoštva, odnosno jednog Boga kao Trojice, koje se već u IV veku iskazuje u učenju kapadokijskih otaca. U izuzetno značajnom filozofsko-teološkom spisu iz VI veka Corpus Areopagiticum, otkrivena je i rekonstruisana trijadološka slika univerzuma. Tu je, pre svega, nasleđena podela teologije na katafatičnu, simboličnu i apofatičnu. Areopagitov sistem satkan je i od relacijskih vrednosti koje vladaju između činova nebeske, crkvene i zemaljske hijerarhije. Ovako zamišljena struktura prepoznata je i pretpostavljena kao teorijski model koji omogućava razumevanje složenih simboličkih slika hrišćanskog sveta. Radi jasnijeg shvatanja, ideja o trojičnosti upoređuje se sa pojmom boromejskog čvora, koga čine tri međusobno isprepletana prstena ili petlje. Logika strukture je takva da se čitava konstrukcija raspada izdvajanjem ili presecanjem samo jednog od prstenova. U Areopagitovoj strukturi tri prstena bile bi tri ličnosti: Otac, Sin, Sveti duh/apofatična, simbolična i katafatična teologija/nebeska, crkvena i zemaljska hijerarhija itd. Na mestu njihovog presecanja je unija, jedinstvenost u mnoštvenosti. Reflektovano na polje arhitekture, trojični princip, kao odnos „tri ipostasi jednosuštnog Boga“, u potpunosti odgovara organskom sklopu sakralne arhitekture vizantijskog hrama i šireg sakralizovanog prostora. Kao predmet ovakvog istraživanja nameću se osnovni aspekti ahitekture — forma, funkcija i konstrukcija. Svaki od aspekata se može posmatrati odvojeno sa svojim kvalitativnim razlikama, ali istovremeno ih je nemoguće razdvojiti jer čine arhitekturu celovitom. Prema tome, posmatrajući ipostasi Trojice, Bog otac, kao prauzrok svih stvari, kao nesaznajan, neopisiv, nesameriv, odgovara funkciji u arhitekturi. Ona ima II transcedentalni karakter: ona je kao ideja savršena, ali se jedino može ispoljiti u konkretnoj formi, što u potpunosti odgovara otkrivanju Boga oca kroz Boga sina — Bogočoveka. Dakle, Otac se ispoljava u Sinu, kao što se funkcija ispoljava kroz formu. Ipostas Hrista odgovara formi u arhitekturi...

Serbian mediaeval architecture, the architecture of the rulers’ endowments, was and still is a materialised testimony to how the Byzantine and consequently the Eastern Christian cultural tradition was being adopted, which brought an opposing blend of the classical and Hebrew thought, expressed in the doctrine of the Holy Trinity. Deciphering the trinity principle constitutes a premise to recognising a theoretical model for interpreting the mediaeval church architecture. Development of the doctrine and solving the questions about God as a Trinity featured the Christian thought dependant on the most significant Judaistic ideas and on those of Late Antiquity, as well as the philosophical conceptual constructions. In analysing and following their adoption, we notice some advancement in solving the questions of understanding the One and the many, i.e. one God as the Trinity, which was made manifest already in the teachings of the Cappadocian Fathers in the 4th century. In a quite significant theological and philosophical text, Corpus Areopagiticum, dating from the 6th century, a triadological image of the universe was discovered and reconstructed. It showed an inherited division of theology into kataphatic, symbolic and apophatic. The Areopagite’s system was constructed of relationships governing among the celestial, ecclesiastical and earthly hierarchies. Thus devised structure was recognised and assumed as a theoretical model which enables understanding of complex symbolic images of the Christian world. For purposes of better understanding, the concept of Trinity is compared with the concept of the Borromean Rings or Knot made of three interlocking circles or loops. The logic of the structure is that cutting any loop or ring frees all the other loops so that no two loops can be directly linked. In the Areopagite’s structure the three rings would be the three Persons: the Father, the Son and the Holy Spirit, the apophatic, symbolic and kataphatic theology, the celestial, the ecclesiastical and the earthly hierarchy, etc. In its centre, inside all the circles, is the Unity, the unity in multiplicity. Mirrored in the field of architecture, the Trinity principle, as a relationship of “Three Hypostases in one Ousia”, completely corresponds to an organic structure of the Byzantine church architecture and a wider ecclesiastic world. As a subject of such research, the basic aspects of architecture come first – the form, the function and the structure. Each of them could be viewed separately, with its qualitative differences, but at the same time, they could not be separated, as they make architecture complete. VI Therefore, considering the hypostases of the Trinity, God the Father, as the First Cause, as unfathomable, indescribable, immeasurable, corresponds to the function in architecture. It is of a transcendental character, perfect as a concept, but can only be manifested in a concrete form, which corresponds completely to revealing God the Father, through God the Son – God-Man. So, God is manifested in the Son, like function is manifested through form. The hypostasis of Christ corresponds to form in architecture...

Konstruktivni postupci u prostornim transformacijama površi jednogranog eliptičkog hiperboloida

The Constructive procedures in the spatial transformations of the surface elliptical hyperboloid of one sheet

Jednograni eliptički hiperboloid (JEH) je najopštiji slučaj površi dvostruke zakrivljenosti, iz porodice pravoizvodnih površi, koja nastaje klizanjem jedne transverzale–generatrise, duž tri direktrise – mimoilazne prave prostora. Disertacija daje odgovor na drugačiji prilaz za postavljanje konstruktivno–geometrijskog okvira površi u prostor. Zadavanjem prostornog šestotemenika – nosača vodilja površi, u različitim varijantama položaja, dva puta po tri izvodnice, iz različitih sistema izvodnica, u novom okviru, razvijen je niz konstruktivnih postupaka, kojima se generiše površ i određuju glavni parametri JEH–a: središte, glavne ortogonalne ose i ravni simetrije, strikciona elipsa i temena i utvrđuju njihove međusobne relacije. Postupcima i naučno zasnovanim metodama projektivne sintetičke geometrije, nacrtne geometrije i alatima kompjuterske grafike, u 3D okruženju (uz upotrebu softvera Auto CAD), teza daje naučni doprinos, kroz rešenja – konstruktivne postupke i teoretske zaključke, u sferi geometrije pravoizvodne površi JEH–a i njenih tipova. U konstruktivnim postupcima prostornih afinih transformacija jednogranih hiperboloida, čije su vodilje upisane u trostrane prizme, utvrđene su invarijante tipova transformacija. Na postavljenim principima inici–rana.su dalja istraživanja relacija opštih i specijalnih tipova JH, u okviru iste familije površi. Analizom svih tipova preseka površi JEH–a sa ravnima, uz uspostavljanje korelacija sa polarnim karakteristikama površi, stvorena je teorijska baza znanja, korisna za praktičnu primenu cele površi, ili njenih isečaka, u postupcima geometrijskog 3D modelovanja. Modeli–strukture, sa zadatim pravilnim matricama, kreirani 3D postupcima, od isečaka pravog ili kosog tipa JEH–a, predstavljaju prototip za široku paletu mogućnosti primene u arhitektonskom oblikovanju krovnih površina ili celih objekata. Sličan, praktično primenljiv, rezultat dao je i deo istraživanja vezan za prodore osnovnih površi drugog stepena i JEH–a. Ovakve kombinacije površi, u postupcima prostornog geometrijskog modelovanja su skup smernica, kao deo inicijative i inspiracije za projektante i graditelje. Rezulatati istraživanja su realizovani i kroz dve aplikacije u softveru Auto CAD (pisanu Auto–lisp programskim jezikom), koja koristi kružne preseke JEH–a, za generisanje žičanog i površinskog modela površi, sa triangulisanom mrežom. Aplikativnost preseka površi po izvodnicama i kružnicama, od značaja je za praksu građenja, koja teži jednostavnosti izvođenja, a da pri tome ne umanjuje estetsku vrednost celokupne forme...

Elliptic hyperboloid of one sheet (EHOS) is the general case of a double curved surface, from the family of ruled surfaces, which generates by ruling of a line–generator along three directing skewed lines, in the space. The dissertation gives an answer to a task – finding a new approach in creating constructive–geometry "boundaries" for this surface in the space settlement. A wide range of constructive procedures, for the determination and mutual relations of the main parameters of EHOS: center of the surface, orthogonal axes, planes of symmetry, minimal section curve (elliptical "throat") and its vertexes came out from the concept of hexagramic spatial "carrier" of two times three directors – skewed lines, in variety of their positions. The employment of procedures and scientifically based methods of Projective Synthetic Geometry, Descriptive Geometry and tools of computer graphics, in 3D surroundings (using Auto CAD software for engineers), are of importance for the scientific contributions of this dissertation, achieved solutions – constructive procedures and theoretical conclusions, in the sphere of geometry of ruled surface EHOS and its types. In the constructive procedures of spatial affine transformations of hyperboloids of one sheet, whose directing lines are "inscribed" in three sided prisms, the invariants of all the types of transformations are determined. Further investigations, concerning relations of general and special types of HOS, inside the same family of surfaces, are initiated by settled principles. The analysis of all the types of sections of EHOS, including the correlation to the polar properties of the surface, resulted with theoretical base, as an input for the practical application of "whole" surface or its fragments, in the procedures of 3D geometrical modeling. The models – structures, above regular patterns (polygons) are created by 3D procedures and tools, using fragments of straight or inclined EHOS. They represent the prototypes for the creative palette of possible architectural shapes of roofs or buildings. Very similar results came out of the part of research, concerning intersections of EHOS and basic second order surfaces (primitives): cone, cylinder and sphere. The "combinations" of such surfaces, in 3D geometrical modeling procedures, offered just some directions, like an initiation and inspiration for the architects and civil engineers...

Uticaj pro- i anti-inflamacijskih faktora na produkciju proteaza vanćelijskog matriksa uPA i MMP9 mišijih makrofaga

Influence of pro- and anti -inflammatory factors on production of extracellular matrix proteases uPA and MMP9 in mouse macrophages

Makrofazi su centralne ćelije imunskog sistema jer poseduju jedinstvene osobine: učestvuju u održavanju fiziološke homeostaze tkiva i organizma, proceni vrste moguće pretnje po organizam i pokretanju adekvatnog imunskog odgovora angažovanjem odgovarajućih komponenti imunskog sistema, ali i njegovom završetku kada više nije neophodan, aktivno učestvuju u obnavljanju oštećenih tkiva i tako omogućavaju povratak homeostaze. Ostvarivanje ovako raznovrsnih uloga omogućuje visoka prilagodljivost ovih ćelija spoljašnjim signalima. Različiti podtipovi makrofaga imaju odlučujuću ulogu u različitim fazama inflamacije. Klasični M1 makrofazi, izloženi dejstvu pro-inflamacijskih citokina, kao što je interferon γ, i/ili produktima mikroorganizama, kao što je lipopolisaharid (LPS), imaju glavnu ulogu u indukciji inflamacije i eliminaciji patogena, dok alternativni M2 makrofazi, u odgovoru na stimulaciju anti-inflamacisjkim/imunoreglatornim citokinima IL-4, IL-13, IL-10, TGF-β ili glukokortikoidima, obustavljaju inflamaciju i ključni su za obnavljanje tkiva i zarastanje rana. Tokom inflamacije odvijaju se značajne promene na nivou proteina vanćelijskog matriksa (ESM, extracellular matrix), koje su strogo regulisane proteazama ESM, čija se produkcija ostvaruje u definisanom prostorno-vremenskom kontekstu. Sekrecijom matriksnih proteaza (MMR) kao što je MMR9, M1 makrofazi učestvuju u degradaciji ESM i razgradnji tkiva, što olakšava ulazak drugih inflamacijskih ćelija u oštećeno tkivo. Istovremeno se pokreće kaskada koagulacije tokom koje se formira privremena struktura ESM od fibrina, koja se kasnije u fazi rezolucije razgrađuje, a ESM se obnavlja. Urokinaza (uPA) je serinska proteaza koja razgradnjom plazminogena u plazmin ostvaruje glavnu ulogu u procesu razgradnje fibrinske mreže. Osim toga, uPA je važan regulator tkivne inflamacije, procesa zarastanja rana i tkivne regeneracije. Otuda je važno rasvetliti molekulski mehanizam regulacije MMR9 i uPA tokom promene polarizacije makrofaga iz M1 u M2. Na osnovu ovih podataka, pretpostavljeno je da se u inflamacijskim makrofazima MMR9 i uPA regulišu na različit način. Da bi se to ispitalo, u eksperimentima je korišćena mišja monocitno- makrofagna ćelijska linija RAW 264.7 kao najčešće korišćeni ćelijski model u studijama koje se bave ispitivanjem biologije makrofaga. Najpre je ispitan uticaj LPS na produkciju uPA od strane ovih ćelija. Rezultati su pokazali da LPS dozno zavisno smanjuje produkciju uPA, a podstiče produkciju MMR9. Analiza LPS signalne transdukcije ukazuje da p38 MAPK učestvuje u smanjenju produkcije uPA. Sa druge strane, prisustvo TGF-β snažno podstiče produkciju uPA a umanjuje produkciju MMR9 i na nivou transkripcije i translacije. Rezultati su dalje pokazali da se uticaj TGF-β na uPA ostvaruje uglavnom kroz Smad3 i ERK1/2 MAPK signalni put, dok bi smanjenje produkcije MMR9 moglo da bude posledica stimulacije formiranja inhibitornog NFκB transkripcionog faktora. Takođe je pokazano da LPS nije u stanju da inhibira kapacitet TGF-β da indukuje uPA. Analize makrofaga poreklom iz kostne srži miševa su potvrdile da M1 i M2 polarizovani makrofazi drugačije eksprimiraju MMR9 i uPA. Kako u većini tumora makrofage odlikuje M2 fenotip, koji doprinosi razvoju tumora, dalja istraživanja su bila usmerena ka efektima paklitaksela (Tah) i estramustin fosfata (ER), anti-tumorskih lekova koji deluju na tubularni citoskelet, na kapacitet TGF-β da indukuje proliferaciju, migraciju i produkciju uPA u RAW 264.7 ćelijama. Pokazano je da i Tah i ER inhibiraju ćelijsku migraciju, ali je samo Tah mogao da u eksperimantalnim uslovima smanji ćelijsku proliferaciju. Uočena redukcija ćelijske migracije korelira sa TGF-β inhibicijom produkcije uPA. Osim toga, Tah i ER su snažno inhibirali aktivnost Smad3 koju je pokrenuo TGF-β. Ovi podaci sugerišu da deregulacija dinamike tubularnog citoskeleta snažno remeti uticaj TGF-β na funkcije makrofaga, što može da ima posledice i na kancerogenezu. Na kraju, ispitan je i efekat glukokortikoida deksametazona (Dex) na produkciju uPA. Intrigantno je da Dex snažno inhibira produkciju uPA u RAW 264.7 ćelijama i da ima adititvno dejstvo sa kapacitetom LPS da umanji produkciju uPA, što bi moglo da bude posledica toga što Dex aktivira r38 signalizaciju. Sa druge strane, inhibitorno dejstvo Dex nije uticalo na kapacitet TGF-β da stimuliše produkciju uPA. Rezultati prikazani u ovoj doktorskoj disertaciji pružaju nove podatke o molekulskom mehanizmu regulacije MMR9 i uPA u mišjim makrofazima, kao i bolje razumevanje regulacije proteaza ESM tokom M1-M2 polarizacije. Imajući u vidu značaj makrofaga u patološkim procesima, i MMR9 i uPA bi mogli da budu ciljni molekuli u terapiji inflamacijskih, malignih i degenerativnih oboljenja.

Macrophages are one of the major effector cells in inflammation. They are responsible for the recognition and processing of foreign materials, activation of the adaptive immune response, regulation of tissue repair/regeneration, and overall homeostasis maintenance. Multiple functions of macrophages are facilitated by their high plasticity in response to environmental or intrinsic signals. Two polarized macrophage modalities are critical in the regulation of different stages of tissue repair after injury. Classical M1 macrophages, exposed to pro-inflammatory cytokines, IFNs and microbial products, such as lipopolysaccharide (LPS), have major role in inducing inflammation and clearing of pathogens, whereas alternative M2 macrophages, in response to IL-4 and IL- 13, IL-10, TGF-β or glucocorticoids, resolve inflammation and are crucial in tissue remodeling and wound healing. In addition to cellular changes, reorganization/remodeling of critical extracellular matrix (ECM) proteins also occurs in injured tissues. During this ECM reorganization, several important ECM proteinases are triggered in a space-temporal fashion through tightly regulated mechanisms. Pro- inflammatory M1 macrophages are able to secrete matrix metalloproteinases, such as MMP9, which help in degradation of the ECM in affected tissue, thus facilitating the recruitment of inflammatory cells to the site of tissue injury. Moreover, an antifibrinolytic-coagulation cascade, that activates clotting and development of a provisional ECM, is also triggered, and later replaced by normal ECM during the resolution of inflammation. The urokinase-type plasminogen activator (uPA) is a serine protease that plays a major role in fibrinolytic processes, where it converts plasminogen to plasmin which further degrades coagulation components. Moreover, uPA is a key regulator of tissue inflammation and wound-healing processes and tissue regeneration, and is also secreted by macrophages. Considering macrophage’s importance in tissue repair, it is crucial to elucidate how MMP9 and uPA are involved in M1-M2 transition, as well as what are the underlying molecular mechanisms. It was hypothesized that MMP9 and uPA are differentially regulated during inflammatory activation of macrophages. To test this hypothesis, immortalized murine macrophage cell line RAW 264.7 was used as a canonical model of immune-competent cells. First the capacity of LPS (as an inductor of M1 state) to regulate uPA production was tested. Results showed that LPS, in a dose-dependent fashion, downregulated uPA production conversely to increased MMP9 production. The analysis of LPS-triggered signaling pathways indicated that LPS-inhibited uPA production was mediated by p38 MAPK. On the other hand, TGF-β (as an inductor of M2 state) greatly induced uPA production parallel to MMP9 downregulation at both protein and transcriptional level. Further results indicated that TGF-β-induced uPA production was mainly mediated by activation of Smad3 and ERK1/2 MAPK signaling transduction, while MMP9 downregulation may be mediated by the inhibition of basal NFκB signaling. In addition, interaction analyses indicated that LPS was not able to inhibit the capacity of TGF-β to induce uPA production. Further analyses which included mouse bone marrow derived macrophages, confirmed differential expression of MMP9 and uPA between M1 and M2 polarized stages. Since in most cancers macrophages residing in tumor microenvironment exhibit M2 phenotype that may maintain tumor growth and expansion, we next investigated the effect of Taxol (Tax) and Estramustine phosphate (EP), anticancer drugs affecting tubulin cytoskeleton, on the capacity of TGF-β to induce cell proliferation, migration and uPA production in RAW 264.7 cells. The results revealed that both Tax and EP inhibit cell migration, although in experimental conditions only Tax was able to decrease cell proliferation. Moreover, the reduction of cell migration was closely related with the inhibition of TGF-β-induced uPA production. Also, both Tax and EP strongly inhibited Smad3 signaling after TGF-β treatment. These data suggested that deregulation of the dynamic of tubulin cytoskeleton strongly interferes with the capacity of TGF-β to regulate mouse macrophages functionality, with putative implications in tumorigenesis. Finally, experiments were performed to determine the effect of the glucocorticoid dexamethazone (Dex) on uPA production. Interestingly, Dex treatment strongly inhibited uPA production in RAW 264.7 cells. Moreover, this effect was additive to the capacity of LPS to downregulate uPA expression, possibly through the activation of p38 MAPK signaling. However, Dex was unable to modify the capacity of TGF-β to stimulate uPA production. Results presented in this thesis, provide new evidence about molecular mechanisms which regulate MMP9 and uPA production in mouse macrophages, as well as allow better understanding of ECM proteases role in M1-M2 macrophages polarization. Furthermore, in the aspect of macrophages, both MMP9 and uPA may be putative therapeutic targets in the treatment of inflammatory, malignant, and degenerative diseases.

Načela moderne u arhitekturi javnih objekata u Beograd, period 1918-1941

Principles of modern architecture in public buildings in Belgrade, the period 1918-1941

Ovaj rad predstavlja prilog proučavanju moderne arhitekture i urbanizma dvadesetog veka u Beogradu. Rad se bavi istraživanjem arhitekture javnih objekata izgrađenim po načelima Moderne kao značajne arhitektonske i šire kulturne pojave u gradu Beogradu i širem jugoslovenskom prostoru u periodu od 1918. do 1941. godine. U ovom periodu sagrađeni su javni objekti sa primenjenim principima Moderne u arhitekturi i urbanizmu. Rad polazi od analize inicijalnih ideja modernog pokreta između dva Svetska rata u Evropi i refleksijama koje je imao na arhitekte koji su preuzimali ovaj model u projektovanju i realizaciji javnih objekata u Beogradu. Sistematizacijom i kritičkom analizom primera realizovanih javnih objekata istražuje se u kojoj je meri Moderna bila zastupljena u Beogradu u arhitektonskoj produkciji tog doba, koji su modeli iz evropskog iskustva preuzimani direktno i indirektno u funkcionalnoj organizaciji, arhitektonskoj volumetriji, konstrukciji i materijalizaciji objekata. Rad obuhvata i hronološku, kritičku analizu najznačajnijih realizovanih objekata u ovom periodu. Istraživanje polazi od stava da su u realizaciji javnih objekata u Beogradu bile prisutne ideje Moderne i avangarde čija su načela bila formirana u kontekstu ključnih istorijskih, društvenih i kulturnih pojava u Evropi i Srbiji. Ovakav stav o problemu istraživanja pretpostavlja uspostavljanje korelacije između načela Moderne i arhitektonskih realizacija domaćih autora kao i teorijsku refleksiju promena koje je Moderna generisala u razvoju Beograda i koje su promovisale novu državu i savremeno društvo.

This work represents a contribution to the study of modern architecture and urbanism of the twentieth century in Belgrade. The paper investigates the architecture of public buildings on the principles of modern architecture as a significant and broad cultural phenomenon in the wider Belgrade and the Yugoslav territory in the period since 1918 untill 1941. During this period public buildings were built with the applied principles of modern architecture and urbanism. The work starts from the initial analysis of the modern movement of ideas between the two World Wars in Europe and the reflections that had the architects who were taking this model in the design and implementation of public buildings in Belgrade. Systematization and critical analysis of realized examples of public buildings to investigate the extent to which modern Belgrade was represented in the architectural production of the time, who were models from European experiences taken over directly and indirectly in the functional organization, volumetry architectural, design and materialization of objects. The work includes a chronological, critical analysis of the most important objects implemented in this period. The research starts from the premise that the preparation of the public buildings in Belgrade were present modern and avant‐garde ideas whose principles were established in the context of key historical, social and cultural phenomenon in Europe and Serbia. This attitude on the issue of research presupposes the establishment of correlation between the principles of modern architecture and implementation by local authors as well as theoretical reflection of changes that is generated in the development of modern Belgrade, which have promoted a new state and modern society.

Uticaj brasinosteroida 24-epibrasinolida na klijanje i rane faze rastenja i razvića različitih hibrida kukuruza (Zea mays L.)

The influence of brassinosteroid 24-epibrassinolide on germination and early stages of growth and development of different maize hybrids (Zea mays L.)

Jedan od važnijih ciljeva moderne poljoprivrede je povećanje efikasnosti (umanjenje troškova proizvodnje, uz istovremeno uvećanje prinosa). Gotovo svi delovi biljke kukuruza se mogu iskoristiti, što je činjenica koja kukuruzu daje poseban ekonomski značaj. Regulatori rastenja i fitohormoni mogu imati povoljan uticaj utiču na povećanje prinosa, razviće i otpornost prema biotskom i abiotskom stresu. Kukuruz je posebno osetljiv na dejstvo nepovoljnih sredinskih faktora u ranim fazama rastenja i razvića, pa tretman regulatorima rastenja može izazvati niz promena u biohemijskim i fiziološkim procesima u klijancima kukuruza, koje bi mogle poboljšati rastenje i otpornost na stresne uslove životne sredine. Brasinosteroidi su prirodni regulatori rastenja biljaka, prisutni u niskim koncentracijama u različitim organima (polen, seme, mlada vegetativna tkiva itd.) biljaka svih sistematskih grupa. Imaju značajnu ulogu u različitim razvojnim procesima kod biljaka kao što je izduživanje ćelija, vaskularna diferencijacija, etiolacija, reproduktivni razvoj itd. Mlada vegetativna tkiva su posebno osetljiva na egzogeno dodate BRs, i u tim tkivima dolazi do intenzivne ekspanzije ćelija. Takođe, BRs posreduju i u odgovoru biljaka na stresne faktore (abiotski, biotski i ksenobiotski). U istraživanjima su korišćena semena hibrida kukuruza ZP 434 i ZP 704, koja su isklijavana u rastvoru 24-EBL-a različitih koncentracija. Za analize su korišćeni klijanci starosti sedam dana. Ispitivan je uticaj egzogeno dodatog 24-EBL-a na rastenje plumule i radikule, kvantifikovan je sadržaj fotosintetičkih pigmenata, mineralnih elemenata i šećera, optimizovana je metoda za ekstrakciju polifenola, uz njihovu karakterizaciju u delovima klijanaca (plumula, radikula, ostatak semena). Detektovana je produkcija reaktivnih kiseoničnih i azotnih vrsta, kako bi se ispitao njihov uticaj na rastenje klijanaca pomenutih hibrida kukuruza. Takođe, sprovedene su kinetičke i termodinamičke analize prilikom isušivanja klijanaca, kako bi se rasvetlili mehanizmi odlaska vode iz delova klijanaca i time analizirali mehanizmi otpornosti klijanaca kukuruza prema toj vrsti stresa. Nađeno je da visoke i niske koncentracije egzogeno dodatog 24-EBL-a imaju različit uticaj na klijavost ispitivanih hibrida kukuruza, kao i da hibridi kukuruza različito reaguju na egzogeno dodati 24-EBL, naročito pri visokim koncentracijama ovog fitohormona. Utvrđeno je da niske koncentracije egzogeno dodatog 24-EBL-a povećavaju vrednosti SVI-II, i da imaju pozitivan efekat na izduživanje plumula klijanaca hibrida ZP 434. Nizak odnos hlorofil a/ hlorofil b u plumulama oba hibrida kukuruza, svedoči o slaboj aktivnosti fotosintetičkog aparata, što ukazuje i da većina asimilata neophodnih za rastenje klijanca dolazi iz semena. Analizom sadržaja mineralnih elemenata, može se zaključiti da klijanci kukuruza ispitivanih hibrida tretirani niskim koncentracijama 24-EBL-a, mogu preživeti u izvesnoj meri dejstvo teških metala zahvaljujući smanjenoj pokretljivosti toksičnih elemenata pre nego što se distribuiraju u plumulu i radikulu. Nađeno je da hibrid ZP 434 sadrži više veoma polarnih fenolnih jedinjenja od hibrida ZP 704. Visoke koncentracije 24-EBL-a inhibitorno deluju na sadržaj polifenola u plumulama hibrida ZP434, a najniža ispitivana koncentracija 24-EBL-a ima inhibitorni efekat na sadržaj polifenola u plumulama hibrida ZP 704. Kod radikule klijanaca ZP 434 hibrida nađeno je da je vrednost prividne energije aktivacije proporcionalna sadržaju trehaloze. Zaključeno je da niže koncentracije 24- EBL-a utiču na uvećavanje sadržaja trehaloze. Ustanovljeno je da kod hibrida ZP 704 prisustvo 24-EBL-a najverovatnije utiče na dehidratacione procese preko visokog sadržaja fosfora u delovima klijanaca, dok je ovaj proces obrnut u organima klijanaca hibrida ZP 434, odnosno kontrolnih uzoraka. Nađeno je da svi organi klijanaca hibrida ZP 704, pokazuju veću energetsku stabilnost u poređenju sa hibridom ZP 434, pri egzogeno dodatom 24-EBL-u tokom procesa dehidratacije. Na osnovu enatalpijsko-entropijske kompezacione teorije, nađeno je da je mehanizam desorpcije vode u ispitivanim hibridima entropijski kontrolisan, pri čemu dobijeni rezultati ukazuju da mikrostruktura klijanaca nije stabilna i da trpi promene tokom dehidratacije. Takođe, na osnovu BET metode i modifikovane BET metode, nađeno je da kijanci hibrida ZP 704 u prisustvu 24-EBL-a lakše odaju vodu za razliku od hibrida ZP 434, kod koga više koncentracije 24-EBL-a izazivaju otežano odavanje vode. Pronađeno je da 24-EBL ima blokirajući efekat na povećanje pritiska širenja.

One of the most important goals of modern agriculture is to increase efficiency (reduction of production costs, with a corresponding increase in yield). Almost all parts of the maize plant can be used, and that is the fact which gives maize a special economic significance. Growth regulators and phytohormones may have a favorable impact on yield improvement, development and resistance to biotic and abiotic stress. Maize is particularly vulnerable to the effects of adverse environmental factors in the early stages of growth and development, and treatment with growth regulators can cause a number of changes in biochemical and physiological processes of maize seedlings which could improve the growth and resistance to stressful environmental conditions. Brassinosteroids are natural regulators of plant growth, and they are present in low concentrations in various organs (pollen, seeds, young vegetative tissue, etc.) at all systematic groups of plants. They have an important role in various developmental processes in plants, such as cell elongation, vascular tissue differentiation, etiolation, reproductive development, etc. Young vegetative tissues are especially sensitive to exogenously added BRs, and in these tissues BRs may induce intensive expansion of cells. Also, BRs can mediate plant responses to stress factors (abiotic, biotic and xenobiotic). Seeds of maize hybrids ZP 434 and ZP 704 were germinated in a solution of different concentrations of 24-EBL. Seven days old seedlings were used for the analyses. The effect of exogenously added 24-EBL on plumule and radicle growth, content of photosynthetic pigments, mineral elements and sugars was analyzed. Also, method for extraction of polyphenols was optimized, along with their characterization in the parts of the seedlings (plumule, radicle, the rest of the seed). Production of reactive oxygen and nitrogen species was rated, in order to examine (with above mentioned methods) their impact on the growth of maize seedlings. Kinetic and thermodynamic analysis during drying of seedlings have been implemented, in order to elucidate the mechanisms of dehydration of seedling parts and thus analyze the mechanisms of resistance of maize seedlings under drought stress. It was found that both high and low concentrations of exogenously added 24- EBL have different effects on the germination of maize hybrids, and that hybrids respond differently to exogenously added 24-EBL, particularly at high concentrations of phytohormones. Low concentrations of exogenously added 24-EBL lead to increase in the value of SVI-II, and have a positive effect on plumule elongation within hybrid ZP 434. The low ratio of chlorophyll a / chlorophyll b, in plumule of two maize hybrids indicates low activity of the photosynthetic apparatus, which means that the most assimilates necessary for seedling growth are coming from the seeds. From the analysis of mineral elements, it can be concluded that the seedlings of maize hybrids treated with low concentrations of 24-EBL may survive areas polluted with heavy metals, due to the reduced mobility of toxic elements before they were distributed to plumulue and radicule. It was found that the hybrid ZP 434 contains more highly polar phenolic compounds than hybrid ZP 704. High concentrations of 24-EBL-a had an inhibitory effect on the content of polyphenols in plumule of hybrid ZP434 and the lowest tested concentration of 24 EBL had an inhibitory effect on the content of polyphenols in plumule of ZP 704. Also, it was found for ZP434 hybrid radicle, that the Ea value is directly proportional to trehalose concentrations. It was concluded that lower concentrations of 24-EBL cause increased values of trehalose. In case of seedlings of ZP704 hybrid, 24- EBL is probably affecting water dehydration by significantly increasing values of phosphorus, comparing to control tests, while process is reverse in a case of ZP434 hybrid. It was found that all the organs of ZP 704 seedlings are having greater energy stability with exogenously added 24-EBL, comparing to the hybrid ZP 434 in the process of dehydration. Based on enthalpy-entropy compensation theory, it has been found that water desorption mechanism in tested hybrids is controlled by entropy, where established results indicate that microstructure of seedlings is not stable and that it suffers from certain changes during dehydration. Also, based on BET and modified BET approaches, it has been found that response of ZP 704 hybrid to 24-EBLs is probably more acceptable than in the case of ZP 434 hybrid. It was found that 24-epibrassinolide has a blocking effect on rapid increase in spreading pressure.

Ispitivanje ranih markera procesa neuroinflamacije i oksidativnog stresa u mozgu transgenog hSOD1 G93A pacova kao eksperimentalnog modela amiotrofične lateralne skleroze

Investigation of early markers of neuroinflammation and oxidative stress in the brain of the transgenic hSOD1 G93A rat as an experimental model for amyotrophic lateral sclerosis

Amiotrofična lateralna skleroza je smrtonosno neurodegenerativno oboljenje koje pogađa motorne i kognitivne domene centralnog nervnog sistema (CNS). Mutacije u genu za Cu, Zn-supeoksid dismutazu (SOD1) uzrokuju 20% familijalnih slučajeva ALS i izazivaju stvaranje unutarćelijskih SOD1 agregata i smanjeno vezivanje Cu i Zn, što dovodi do patoloških unutarćelijskih promena i neurodegeneracije. Istraživanja na post mortem tkivu ALS pacijenata i životinjskim modelima bolesti su istakla glijalnu aktivaciju, poremećenu homeostazu prelaznih metala, oštećenje krvno-moždane barijere (KMB) i povećanu proizvodnju reaktivnih kiseoničnih vrsta kao glavne patološke faktore u bolesti. Cilj ove studije je bio da se ispitaju navedene patološke promene u motornim (moždano stablo) i ne-motornim (hipokampus) centrima u mozgu transgenog hSOD1 G93A pacovskog modela ALS, u presimptomatskom i simptomatskom stadijumu bolesti, kako bi se detaljnije proučili ovi patofiziološki mehanizmi i identifikovale rane promene u bolesti. Prisustvo i vremenski sled aktivacije i morfoloških promena mikroglije i astrocita, kao i akumulacija i distribucija SOD1 proteina u ovim ćelijama su ispitani naprednim 3D modelovanjem i analizom kolokalizacije na slikama dobijenim konfokalnom laserskom skenirajućom mikroskopijom. Rezultati su pokazali glijalnu proliferaciju i progresivno nakupljanje SOD1 u tkivu u oba ispitivana moždana regiona hSOD1 G93A pacova, koji počinju već u presimptomatskoj fazi. U moždanom stablu ovih životinja aktivacija astrocita je bila prisutna pre početka simptoma bolesti, i bila je praćena aktivacijom mikroglije. Takođe, u moždanom stablu astrociti su pokazali progresivnu akumulaciju SOD1 proteina u ćelijskom telu i nastavcima, dok su nivoi SOD1 u mikrogliji bili smanjeni a njena distribucija ograničena na distalne ćelijske nastavke. U hipokampusu hSOD1 G93A pacova astrociti su pokazali isti reaktivni profil kao onaj uočen u moždanom stablu, dok je morfologija mikroglije bila nepromenjena. U ovom regionu mozga oba tipa glijalnih ćelija su pokazala akumulaciju SOD1 proteina u ćelijskom telu...

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder affecting both the motor and cognitive domains of the central nervous system (CNS). Mutations in the Cu,Zn-superoxide dismutase (SOD1) cause 20% of familial ALS and provoke formation of intracellular SOD1 aggregates and copper and zinc unbinding, leading to pathological intracellular changes and neurodegeneration. Post mortem and animal model studies indicate glial activation, perturbed transition metal homeostasis, blood-brain barrier (BBB) disruption and elevated production of reactive oxygen species as major contributors to disease pathology. The aim of this study was to investigate the above mentioned pathological changes in motor (brainstem) and non-motor (hippocampus) brain centers of a transgenic hSOD1 G93A rat model of ALS, in presymptomatic and symptomatic stages of the disease, in order to further clarify these pathophysiological mechanisms and to identify early changes in the disease. The presence and the timeline of activation and morphological changes of microglia and astrocytes, as well as their intracellular accumulation and distribution of SOD1 were investigated by advanced 3D modeling and colocalization analysis of images obtained by confocal laser scanning microscopy. Results revealed glial proliferation and progressive tissue accumulation of SOD1 in both brain regions of hSOD1 G93A rats starting already at the presymptomatic stage. In the brainstem of these animals astrocytes activation occurred before disease symptoms onset, and was followed by activation of microglia. Additionally, in the brainstem astrocytes demonstrated progressive SOD1 accumulation in the cell body and processes, while microglial SOD1 levels were reduced and its distribution limited to distal cell processes. In the hippocampus of hSOD1 G93A rats astrocytes exhibited an identical reactive profile as the one observed in the brainstem, while microglial morphology was unchanged. In this brain region both glial cell types exhibited SOD1 accumulation in the cell body...

Prostorna organizacija stambenih zgrada građenih u Beogradu od 1947. do 1980. godine u prefabrikovanim industrijalizovanim sistemima

Spational organization of residential housing realised in Belgrade, in period between 1947th and 1980th year, in precast industrial systems

Disertacija je nastala kao nastavak istraživanja i magistarske teze “Valorizacija modernističke baštine, na primeru stambene arhitekture Beograda od 1947. do 1967. godine u Beogradu”, odbranjene na Arhitektonskom Fakultetu Univerziteta u Beogradu, 2006.godine. Tokom dugogodišnjeg istraživanja naučne literature u oblasti tehnologije, projektovanja, istorije moderne arhitekture Beograda, kao i višegodišnjeg posmatranja i rada u oblasti projektovanja stambenih i drugih objekata, izvođenja, rekonstrukcijama zgrada, često rađenih u prefabrikovanim sistemima, napravljena je selekcija materijala, i izvršena valorizacija arhitektonskih ostvarenja, koje svojim karakteristikama, zasigurno obeležavaju epohu u kojoj su nastala. Košićena je istoriografska metodologija istraživanja. U radu se postavljaju hipoteze, ciljevi istraživanja, i sažeto predstavlja veoma kompleksna epoha koja je predmet istraživanja, kroz socio-kulturološki, tehnološki, i ekonomski aspekt. Vrednovanje moderne stambene arhitekture Beograda zasnovano je na metodološkim principima koji se, čini se svojom uspešnom primenom u praksi u mnogim zemljama, nameću kao relevantni i oni su dali smernice za formiranje odgovarajućih kriterijuma za vrednovanje. Svojevrsni doprinos rada je u razvoju metodologije vrednovanja moderne kulture uopšte. Posebna pažnja se poklanja društvenim okolnostima u kome je takva masovna produkcija nastala i razvijana. Analiza drušvenih kretanja, političkih prilika, kao i demografskih pokazatelja, migracija stanovništva, i privrednog razvoja ukazuje na određene vrednosti koje je proučavani period negovao i promovisao. Analiza stambenih politika pokazala je da su one uzročno-posledično povezane sa razvojem sistema VI prefabrikovane gradnje, generalno idejama masovne stambene izgradnje, i realizacije velikih stambenih naselja. Kroz rad se razmatra nekoliko važnih pitanja u oblasti stambene izgradnje – da li postoji mogućnost definisanja tj. izdvajanja oblika koji reprezentuju određeni politički metod; zatim, da li bi moderna arhitektura izgledala drugačije da su stambene politike funkcionisale na drugi način; i – na koji su način prefabrikovani industrijalizovani sistemi upotrebljivi u savremenoj socijalnoj stambenoj izgradnji. Pored toga, u radu se razmatra tema – da li se za određene tipove naselja i određene društvene grupe može vezati konkretan tip prefabrikovanih sistema stambene gradnje. U jednom delu rada, daje se pregled tadašnjih tendencija u savremenim umetničkim praksama, pre svega u srodnim oblastima vajarstva, slikarstva i drugih likovnih umetnosti, ali i u masovnoj kulturi, koju su konzumirali korisnici stambenih prostora. U radu se povlače paralele i nudi se odgovor na pitanja da li se u prefabrikovanoj izgradnji prepoznaju oblici savremenih umetničkih izraza. Zahvaljujući razmeni informacija i otvaranju prema svetu, neosporan je izvesni uticaj iz inostranstva, koji je na različite načine dolazio u tadašnju Jugoslaviju. Uticaj, praktično razmena informacija vršena je preko razmene literature, naučnih i stručnih knjiga i časopisa, prevođenja tekstova stranih autora, kao i povremenih izložbi savremenih slikara i arhitekata koje su organizovane u Beogradu. Pored toga, nisu bila retka stručna usavršavanja i studijska putovanja u inostrane akademske centre. Stambene politike nisu imale kontinuitet, i nisu bile dugog trajanja, nego su se vremenom menjale, te je u razmatranom periodu nekoliko puta došlo do uzročno- posledičnih promena na drugim poljima. To je uticalo na promene odnosa između aktera – investitora, građevinskih direkcija, veliih preduzeća, i kupaca (radnih organizacija, sindikata, lokalnih i republičkih vlasti i institucija sistema). U radu se razdvajaju uticajni faktori i pravi paralela sa lokacijama stambenih naselja kroz analizu aktera – onih koji su gradili i – onih za koga su naselja bila projektovana i građena. Tako se dolazi do izvesnih zaključaka odnosa šire centralne zone prema perifernim naseljima Beograda i tumači tema socijalne segregacije. VII Zahvaljujući masovnoj stambenoj izgradnji, i velikom broju sistema i primera, urađena je i predstavljena detaljna tipologija, s tim da se na pojedine naučne studije, konkurse, i kataloge posebno skrenula pažnja, jer su u izvesnom smislu predstavljali prekretnicu u praksi. Ključni deo rada predstavlja deo posvećen valorizaciji izabranih primera. U delu o tipologijama, uvodi se diskusija o slobodnim prostorima gde su na nivou novoformiranih, ali i starih formiranih gradskih blokova poništavane parcele. Ovaj urbanistički pristup nedvosmisleno se povezuje sa društvenim odnosom prema temama i problemima svojine, i odnosa privatnog i javnog prostora. U tekstu se objašnjava pojava – kako i zašto varira odnos jedne urbanističke strukture, i – kako prefabrikacija utiče na urbanističku celinu, da li se broj varijantnih rešenja organizacije bloka povećava ili smanjuje. Na nivou arhitekture stambene zgrade, ovaj uticaj se razmatra detaljnije, i objašnjava se kako prefabrikacija utiče na odnos funkcionalnih celina i zona u stambenoj jedinici. Takođe, posmatraju se i odnosi veličina kuhinja i broja soba, i analiziraju tipska rešenja kuhinjsko-kupatilskog bloka u odnosu na generalnu predloženu tipologiju. Daje se pregled – u kom periodu je koji tip stambenih jedinica bio dominantan. Povlači se izvesna paralela sa tradicionalnom organizacijom beogradskog stana karakterističnog po centralnoj prostoriji. Pored upotrebne vrednosti koja je neosporna, govori se o pitanjima spomeničke valorizacije, jer je jedan od dugoročnih ciljeva rada na ovoj temi, upravo inicijativa za institucionalnom zaštitom što više reprezentativnih primera epohe moderne stambene arhitekture u Beogradu. Daju se obrazloženja izabranih primera. U istraženoj objavljenoj literaturi nije mnogo razmatrana tema kriterijuma za valorizaciju moderne i savremene arhitekture, te taj deo rada predstavlja izvesni naučni doprinos, kao rezultat obavljenih istraživanja. Zaključak doktorske disertacije sumira sve osnovne elemente i rezultate obimnog istraživanja. Daje se uvid u osnovne probleme, analiziraju vrednosti, naglašavaju karakteristični primeri i ističu rezultati. VIII Za potrebe doktorske disertacije, u međuvremenu napravljen je opširni katalog, detaljni pregled skoro svih arhitektonskih sklopova(osnova tipskog sprata) i stambenih jedinica izgrađenih u Beogradu u prefabrikovanim industrijalizovanim sistemima (uključujući i potpuno prefabrikovane i delimično prefabrikovane tj.kombinovane realizacije). U okviru zaključnih razmatranja, razrađuje se koncept remodelacije. Takođe, napredak tehnologije i programskih jezika omogućava izradu softvera, koji putem kombinatorike dokazuju veliki broj mogućnosti prostornih organizacija stambenih zgrada, uprkos malom broju definisanih elemenata, zadatih parametrima industrijalizovanih prefabrikovanih sistema. U prilogu rada nalaze se reprezentativne fotografije, i katalozi koji šematski i grafički prikazuju najkarakterističnije primere arhitektonskih sklopova (osnova tipskog sprata) i stambenih jedinica izgađenih u prefabrikovanim industrijalizovanim sistemima.

The dissertation was created as an extension of the research and Magister's thesis "Valorization of the modernist heritage, on the example of residential architecture in Belgrade at 1947 to 1967 in Belgrade", defended at the Faculty of Architecture, University of Belgrade, 2006. During this research of the scientific literature in the field of technology, design, history of modern architecture of Belgrade, as well as many years of observation and work in the design of housing and other buildings, construction , building renovations often done in precast systems, the selection of materials is made, along with the evaluation of architectural achievements, which showcased certainly mark the epoch in which they occurred. Socio – historical research methodology was used. The paper raises hypothesis, research objectives, and summarize a very complex era that is the subject of research, and its socio – cultural, technological, and economic aspects. Evaluation of modern residential architecture is based on the methodological principles which are, as it seems fro their application in practice in many countries, imposed as they are relevant and provided guidance for the establishment of appropriate evaluation criteria. This paper represents genuine contribution to the development of a methodology for evaluation of modern culture. Special attention is paid to the social circumstances in which such mass production emerged and developed. Analysis of social movements, political situation, as well as demographics, migration, and economic development points to a certain value which is studied period fostered and promoted. The analysis of housing policy has shown that they are the cause and effect associated with the development of prefabricated building systems, the general idea of mass housing construction, and implementation of large housing estates. X Throughout the paper discusses several important issues in the field of residential construction – whether it is possible to define that extraction forms that represent a political method, then, would the modern architecture looked different to the housing function in a different way, and – usage of precast industrialized systems in modern social housing. In addition, the paper discusses the issue – whether certain types of settlements and certain social groups can be bounded to the specific type of prefabricated housing construction system. In one part, this paper gives an overview of that period tendencies in contemporary art practices, particularly in the related fields of sculpture, painting and other visual arts, but also in mass culture, which are consumed by the customers of housing. In this paper we draw parallels and provides an answer to the question, whether the prefabricated building recognized forms of contemporary artistic expression. Thanks to the exchange of information and opening to the world, a certain undeniable influence from abroad, which in many ways came to what was then Yugoslavia. Influence, in fact exchange of information, is carried out through the exchange of literature, scientific and professional books and journals, translated texts of foreign authors, as well as temporary exhibitions of contemporary artists and architects organized in Belgrade. In addition, professional specialization and study tours to foreign academic centers haven’t been rare thing. Housing policies did not have continuity, and they were not of long duration, but they have changed over time, and in the period considered several times for changes. This led to changes in the relationship between the stakeholders – the investors, the building directorate, large enterprises and customers (labor organizations, trade unions, local and national government and state institutions). The paper separates the influencing factors and parallels with the location of residential areas through analysis of stakeholders – those who have built and – those for whom the settlements were designed and built. So we come to certain conclusions about relations beyond the central zone to suburbia settlements in Belgrade and explains topics of social segregation. XI Thanks to the massive housing construction, and a large number of systems and examples, detailed typology was made and presented, with the proviso that the individual scientific studies, competitions, and catalogs in particular drew attention because they are in some sense a practical turning point. A key part of the work is devoted to evaluation of selected examples. In the section on typologies, the discussion on open areas was introduced, where the level of newly formed, and the old established city blocks erased plot. This urban approach clearly associated with social attitudes toward the topics and issues of ownership, and between the private and public space. The article explains the phenomenon – how and why the relationship varies one urban structures, and – as an affect of the prefabrication on the urban whole, whether the number of alternative solutions of the block increases or decreases. At the architecture of the building level, this impact is considered in more details, and explains how the prefabrication affects the relationship between functional areas and zones in a dwelling. Also, the relations between size kitchen and room number are observed, and standard solutions of bathroom-kitchen block analyzed in relation to the proposed general typology. Which type of housing unit was dominant during which time is reviewed. Some parallels with the traditional organization of Belgrade apartment typical for the central area were withdrawed. In addition to its value in use, which is undeniable, this paper talks about the issues of cultural valorization, since one of the goals of long-term work on this subject, it is an initiative of the institutional protection of representative examples of the modern era in residential architecture in Belgrade. Give the reasons for selected examples. Explored in the published literature is not much discussed topic evaluation criteria for modern and contemporary architecture, and this part is a certain scientific contribution as a result of the conducted research. A dissertation conclusion summarizes the main elements and the results of extensive research. It gives an insight into the basic problems, analyze the values, and highlights the typical examples and the results. XII For the purposes of a dissertation, in the meantime, an extensive catalog was made, a detailed review of almost all architectural components (base typical floor) and residential units built in Belgrade in industrialized precast systems (including complete and partial prefabricated – combined realizations). In the concluding remarks, the concept of remodeling was developed. Also, the advance of technology and programming languages allow the development of software, which, through combinations, proves a number of features of spatial organization of residential buildings, despite the small number of defined elements, given the parameters of industrialized precast systems. In the appendix are representative photos, and catalogs schematically and graphically show the most characteristic examples of architectural components (base typical floor) and residential units built in industrialized precast systems.

Moderno i vernakularno u procesu urbane transformacije Banjaluke

The modern and the vernacular in urban transformation of Banja Luka

Ispitivanje urbane istorije malog grada pripada do sada nedovoljno pokrivenom istraživačkom polju u oblasti arhitekture i urbanizma. Istraživanje relacija prema naslijeđenom i prošlosti u procesu urbane transformacije Banjaluke otkriva jedan od oblika modernog iskustva koji se odvija izvan metropola, na periferiji nekadašnjih velikih imperija, u kontekstu u kom se nikako ili sporo uspostavljaju moderni društveno–ekonomski uslovi (u terminologiji Maršala Bermana „modernizam nerazvijenosti“), i doprinosi razumijevanju složenosti i pluralnosti modernog i modernizacije, viđenih iz perspektive istraživača – arhitekte. Problem istraživanja je specifična modernost malog grada u kontekstu zakašnjelih i usporenih transformacija načina života, sagledana u domenu njegove arhitekture i urbanizma. Kroz mnogostruke, heterogene i ambivalentne relacije novog i naslijeđenog, u malom gradu se razvija kompleksna gramatika fizičkog i mentalnog pripadanja i dolazi do novih načina razmišljanja kojima se oblikuje otpor destruktivnim aspektima modernizacije. Predmet istraživanja je Banjaluka u procesy urbane transformacije u periodu od kraja XIX vijeka do 1975. godine. U vezi sa istraživanjem problema kulturnog identiteta u kontekstu BiH danas, u domenu arhitekture i urbanizma i urbane istorije povećano je zanimanje i javljaju se nove interpretacije tema povezanih sa periodom austrougarske uprave ili periodom Vrbaske banovine. Ovaj rad se uključuje u tu debatu ispitujući relacije prema naslijeđenom i prošlom u arhitekturi i urbanizmu u manjoj sredini, daleko od uticaja velikih urbanih centara. Naime, u dosadašnjim radovima o arhitekturi i urbanizmu u BiH interesovanje je usmjereno na zbivanja na arhitektonskoj sceni Sarajeva, dok o arhitekturi drugih gradova, uključujući i Banjaluku, nema sistematskih studija...

The examination of urban history of small towns belongs to the research area of architecture and urbanism that has not been fully covered yet. The research on the relations towards the past and the inherited in the process of urban transformation of Banjaluka reveals one of the forms of modern experience that develops outside the metropolis, at the periphery of former grand empires, in the context where modern socio–economic conditions are established slowly or not at all (modernism of underdevelopment, as defined by Marshall Berman), and contributes to the understanding of complexity and plurality of the modern and the modernization, as seen from the perspective of an architect – researcher. The research problem is the specific modernity of a small town in context of belated and slow transformations of the ways of life, seen in its architecture and planning. A complex grammar of physical and mental belonging has been developed through manifold, heterogeneous and ambivalent relations of the new and the inherited in the small town, and new ways of thinking have emerged, that have shaped the resistance to the destructive aspects of modernization. The subject of analysis and interpretation is Banjaluka in the process of transformation between the end of 19th century and 1975. In relation to the research on the issues of cultural identity in the present context of Bosnia and Herzegovina, interest has been raised and new interpretations have emerged in architecture and urban history for the matter related to the Austro–Hungarian or Vrbas Banate periods. This work joins the debate, by examining relations with the inherited and the past in architecture and urbanism of a small town, far away from the influence of big urban centers. It should be mentioned that the focus of interest in the existing works on architecture and urbanism of Bosnia and Herzegovina has been on the architectural scene of Sarajevo, while there are no systematic studies about the architecture of other cities, including Banjaluka...

Ekološka studija invazivnih biljnih vrsta u ramsarskim područjima Vojvodine

Ecological study of invasive plants in the Ramsar sites of Vojvodina

Invazivne vrste su globalno prepoznate kao jedan od najvažnijih faktora ugrožavanja biodiverziteta, a ekosistemi vlažnih područja su posebno podložni biološkim invazijama. Vlažna područja su generalno veoma fragilna i ugrožena zbog čega im je, Ramsarskom konvencijom, dodeljen poseban konzervacioni status na globalnom nivou. Podaci o invazivnim biljkama i njihovim zajednicama u zaštićenim područjima su vrlo oskudni, a Ramsarskom konvencijom nisu definisane odredbe koje regulišu zaštitu vlažnih staništa od invazivnih vrsta. Zbog svega, osnovni ciljevi ove disertacije su utvrđivanje prisustva i distribucije 18 odabranih invazivnih biljnih vrsta i njihovih zajednica, kao i procena invazibilnosti različitih staništa u šest ramsarskih područja severne Srbije, sa kritičkom analizom pravnih aspekata primene Ramsarske konvencije u pogledu kontrole invazivnih vrsta. Istraživanja su obavljena u periodu 2011.- 2015. godina. Napravljeno je ukupno 669 fitocenoloških snimaka sa 481 biljnim taksonom unutar 15 tipova staništa. Kao najzastupljenije, izdvojile su se vrste: Aster lanceolatus, Amorpha fruticosa, Bidens frondosa, Echinocystis lobata i Asclepias syriaca. Na osnovu udela invazivnih vrsta u opštoj pokrovnosti (indeks D%), najviša invazibilnost je utvrđena na prostoru Carske bare i Koviljsko-petrovaradinskog rita, u strukturno složenijim i vlažnijim staništima (C3.2, C3.5, F9.1, F9.3, G1.1), kao i narušenim ili veštačkim staništima (G1.C, I1.5). Najniža invazibilnost je utvrđena za područje Slanog Kopova, kao i za suvlja staništa (D5.2, E1.2, G1.A). Sintaksonomskom analizom su definisane tri grupe invazivnih zajednica sa 12 diferenciranih asocijacija. Rezultati upućuju na odgovorniji, efikasniji i multidisciplinarni pristup u zaštiti istraživanih područja.

Invasive species are globally recognized as one of the most important factor for biodiversity loss and wetland ecosystems are particularly susceptible to biological invasions. Wetlands are generally very fragile and vulnerable, which is the reason why they have special global conservation status by the Ramsar Convention. Data on invasive plants and their communities in protected areas are very scarce, and the Ramsar Convention does not prescribe provisions for the protection from invasive species. Hence, the main objectives of this dissertation are determination the presence and distribution of 18 selected invasive plant species and their communities, as well as the assessment of invasibility of different habitats in six Ramsar sites of the northern Serbia, with a critical analysis of the legal aspects of implementation of the Ramsar Convention regarding the control of invasive species. Research was carried out in the period 2011- 2015. A total of 669 phytocenological relevés were made with 481 plant taxa within 15 habitat types. The following species were the most abundant: Aster lanceolatus, Amorpha fruticosa, Bidens frondosa, Echinocystis lobata and Asclepias syriaca. Based on the coverage index (D%), the highest invasibility was determined in the area of Carska bara and Koviljsko-Petrovaradinski Rit, in the more structured and humid habitats (C3.2, C3.5, F9.1, F9.3, G1.1), as well as disturbed or artificial habitats (G1.C, I1.5). The lowest invasibility was determined for Slano Kopovo, and in more arid habitats (D5.2, E1.2, G1.A). The syntaxonomic analysis identified 3 groups of invasive communities with 12 differentiated associations. The results indicate a more responsible, efficient and multidisciplinary approach to the protection of the research areas.

Identifikacija molekularno-genetičkih markera patogeneze akutne mijeloidne leukemije

Identification of molecular genetic markers of acute myeloid leukemia pathogenesis

Akutna mijeloidna leukemija (AML) predstavlja grupu hematoloških malignih oboljenja koju karakteriše heterogeni genetički i epigenetički profil. AML sa normalnim kariotipom (AML-NK) predstavlja najveću podgrupu u okviru AML, koju odlikuje intermedijarni rizik. Međutim, samo 40% obolelih od AML-NK preživljava duže od 5 godina, pa je neophodno otkriti molekularno- genetičke markere značajne za prognozu i praćenje toka bolesti, rano otkrivanje minimalne rezidualne bolesti (MRB) i izbor odgovarajuće terapije, kako bi bila omogućena bolja stratifikacija pacijenata sa AML-NK. Prvi cilj ove studije je analiza mutacionog profila dečje (dAML) i adultne (aAML) koja će doprineti rasvetljenju patogeneze mijeloidnih leukemija. Drugi cilj predstavlja određivanje nivoa ekspresije WT1, EVI1, BAALC i MN1 gena, kao potencijalnih molekularno-genetičkih markera značajnih za AML-NK. Analizirani su neselektovani DNK uzorci 20 dAML i 20 aAML pacijenata, primenom metode targetovanog sekvenciranja nove generacije (eng. „targeted next generation sequencing“ – NGS), TruSeq Amplicon - Cancer Panel (TSACP) esejem, koji omogućava detekciju somatskih mutacija u 48 gena uključenih u kancerogenezu. U drugom delu ove studije nivo ekspresije gena WT1 (104 pacijenta), EVI1 (104 pacijenta), BAALC (111 pacijenata) i MN1 (111 pacijenata) u mononuklearnim ćelijama koštane srži AML-NK pacijenata i zdravih pojedinaca analiziran je real- time PCR metodom. Rezultati su pokazali da AML sadrži relativno mali broj genetičkih promena, u proseku samo 3 mutacije po pacijentu kod aAML i dAML grupe pacijenata. Zastupljenost najčešćih mutacija u pojedinačnim genima, koji su povezani sa patogenezom AML, razlikovala se kod dAML i aAML: IDH1 (0% kod dAML, 5% kod aAML), IDH2 (0% kod dAML, 10% kod aAML), NPM1 (10% kod dAML, 35% kod aAML). Takođe, prisustvo mutacija u 4 gena (JAK3, ABL1, GNAQ i EGFR) koji kodiraju tirozin kinaze ili proteine koji su udruženi sa tirozin kinazama detektovano je isključivo kod dAML pacijenata, dok je kod 5 gena (IDH1, APC, HNF1A, GNAS i SMARCB1) koji su uključeni u metilaciju i modifikaciju histona prisustvo mutacija utvrđeno isključivo kod aAML pacijenata. Grupe pacijenata sa visokim nivoom ekspresije WT1 ili BAALC ili MN1 gena imale su kraće trajanje kompletne remisije (KR) i kraće ukupno preživljavanje (UP). Stoga se zaključuje da visok nivo WT1, visok nivo BAALC i visok nivo MN1 ekspresije, svaki za sebe, predstavlja nepovoljan prognostički faktor. Pokazano je da pacijenti sa povišenim nivoom ekspresije EVI1 imaju povećan rizik od pojave relapsa bolesti, ali bez uticaja na dužinu trajanja KR i UP. Razlika u mutacionom profilu između dAML i aAML pacijenata ističe postojanje razlike u patogenezi AML kod ove dve grupe bolesnika, a nalaz da AML kod obe grupe pacijenata sadrži relativno mali broj mutacija u odnosu na druge malignitete čini ovu bolest pogodnom za primenu molekularne ciljane terapije. Naši rezultati govore u prilog da ekspresija WT1 predstavlja dobar molekularni marker za praćenje MRB, odgovora na terapiju i predikciju relapsa bolesti, a da ekspresija BAALC predstavlja stabilan molekularni marker pogodan za praćenje MRB i odgovora na terapiju kod AML-NK. Ekspresija MN1 gena predstavlja molekularni marker pogodan za praćenje odgovora na inicijalnu terapiju, ali nije stabilan marker za praćenje MRB kod AML-NK. Ova studija je ukazala na različitu molekularnu osnovu dAML i aAML, kao i na potencijalne targete za ciljanu terapiju. Ekspresije WT1, BAALC i MN1 predstavljaju nove molekularne markere značajne za prognozu AML-NK.

Acute myeloid leukemia (AML) is a group of haematological malignancies characterized by a heterogeneous genetic and epigenetic profile. AML with normal karyotype (AML-NK) represents the largest subgroup within the AML, characterized by an intermediate risk. However, only 40% of patients with AML-NK survive for more than 5 years, so it is necessary to detect molecular genetic markers significant for prognosis and disease monitoring, early detection of minimum residual disease (MRD), and the selection of appropriate therapy in order to enable better stratification of patients with AML-NK. The first aim of this study is the analysis of the mutation profile of childhood (cAML) and adult (aAML) AML which will contribute to the clarification of the pathogenesis of myeloid leukemia. The second aim is to determine the level of expression of WT1, EVI1, BAALC and MN1 gene, as potential molecular genetic markers relevant to AML-NK. Unselected DNA samples of 20 cAMLs and 20 aAML patients were analyzed using the targeted next generation sequencing (NGS) method, on the TruSeq Amplicon-Cancer Panel (TSACP), which enables the detection of somatic mutations in 48 genes involved in carcinogenesis. In the second part of this research, the level of expression of the genes WT1 (104 patients), EVI1 (104 patients), BAALC (111 patients) and MN1 (111 patients) was analyzed in mononuclear bone marrow cells of AML-NK patients and healthy individuals by the real-time PCR method. The results showed that the AML contains a relatively small number of genetic changes, on average only 3 mutations per patient in the aAML and cAML group. The incidence of the most common mutations in individual genes, which are associated with AML pathogenesis, differed in cAML and aAML: IDH1 (0% in cAML, 5% in aAML), IDH2 (0% in cAML, 10% in aAML), NPM1 (10% in cAML, 35% in aAML). Also, the presence of mutations in 4 genes (JAK3, ABL1, GNAQ, and EGFR) that encode tyrosine kinases or proteins that are associated with tyrosine kinases were detected exclusively in cAML patients, while in 5 genes (IDH1, APC, HNF1A, GNAS and SMARCB1) involved in methylation and histone modification, the presence of mutations was found exclusively in aAML patients. Groups of patients with high level of expression of WT1 or BAALC or MN1 gene had a short duration of complete remission (CR) and shorter overall survival (OS). It is therefore concluded that a high level of WT1, a high level of BAALC and a high level of MN1 expression, each for itself, is an unfavorable prognostic factor. Patients with elevated levels of EVI1 expression have been shown to have an increased risk of relapse of the disease, but without affecting the duration of CR and OS. The difference in the mutational profile between cAML and aAML patients highlights the existence of a difference in AML pathogenesis in these two groups, and the finding that AML in both groups of patients contains a relatively small number of mutations compared to other malignancies makes this disease suitable for the application of molecular targeted therapy. Our results suggest that expression of WT1 represents a good molecular marker for MRD monitoring, response to therapy and prediction of relapse, and that expression of BAALC is a stable molecular marker suitable for monitoring MRB and response to therapy in AML-NK. MN1 expression is a molecular marker suitable for monitoring response to initial therapy, but it is not a stable marker for monitoring MRD in AML-NK. This study pointed to an existence of a different molecular basis of cAML and aAML, and to potential targets for targeted therapy. Expressions of WT1, BAALC and MN1 represent new molecular markers significant for the prognosis of AML-NK.

Tresave Srbije klase Scheuchzerio-Caricetea fuscae (Nordh. 1936) R. Tx. 1937. - florističko vegetacijske karakteristike, ugroženost i zaštita

Fens of the class Scheuchzerio-Caricetea fuscae (Nordh. 1936) R. Tx. 1937. in Serbia - floristic and vegetation characteristics, threats and protection

Tresave Srbije i Balkanskog poluostrva predstavljaju južne granice distribucije tresava u Evropi. Tresavska vegetacija, odnosno vegetacija u sintaksonomskom smislu obuhvaćena je klasama Oxycocco-Sphagnetea Br.-Bl. et Tüxen ex Westhoff et al. 1946 i Scheuchzerio-Caricetea fuscae (Nordh. 1936) R. Tx. 1937., od kojih je na teritoriji Republike Srbije zabeležena samo vegetacija klase Scheuchzerio-Caricetea fuscae. Ova vegetacija je prestavljena većim brojem malih i veoma malih tresava brdsko planinskih predela i osnovni je predmet ove doktorske disertacije. Ciljevi ove disertacije su višestruki: uobličavanje opših teorijskih postavki i definicija tresava i treseta, njihova geneza, ekologija, distribucija i značaj; sistematizacija svih relevantnih publikovanih i nepublikovanih podataka; analiza postojećih klasifikacionih sistema; sintaksonomske i florističke analize prikupljenih podataka o tresavama Srbije; utvrđivanje aktuelnog stanja i faktora ugrožavanja tresavske flore i vegetacije; analiza nacionalne i međunarodne legislative iz oblasti zaštite prirode; analiza do sada zaštićenih tresavskih područja u Srbiji sa uočenim prednostima i slabostima zaštite. Na bazi komparativnih analiza svih dostupnih vegetacijskih snimaka klase Scheuchzerio-Caricetea fuscae na nivou Evrope, uključujući Srbiju (Peterka et al. 2016 submitted), urađen je predlog revizije nacionalne sintaksonomije tresavske vegetacije pri čemu su sve biljne asocijacije svrstane u dva reda: Caricetalia fuscae W. Koch 1926 emend. Nordhagen 1937 i Caricetalia davallianae Br.-Bl. 1950, i 4 sveze: Caricion fuscae Koch 1926, Sphagno-Caricion canescentis Passarge (1964) 1978, 3arthecion scardici Horvat ex Lakušić 1968 i Caricion davallianae Klika 1934. Sveza Sphagno (recurvi)- Caricion canescentis po prvi put je konstatovana za Srbiju dok je sveza Caricion davallianae prethodno samo literaturno navođena, bez konkretnih publikovanih fitocenoloških snimaka. Novoopisane biljne asocijacije imenovane su radno, i tek predstoji njihova definitivna obrada radi publikovanja. Unutar sveze Caricion fuscae svrstano je 19 asocijacija. Dijagnostičke vrste na nivou Srbije su: Carex nigra, Eriophorum angustifolium, Carex echinata, 3ardus stricta, Sphagnum subsecundum. Sveza Sphagno-Caricion canescentis predstavljena je sa 4 asocijacije. Dijagnostičke vrste na nivou Srbije su: Carex rostrata, Equisetum fluviatile, Menyanthes trifoliata, Polytrichum commune, Sphagnum fallax, Sphagnum flexuosum, Sphagnum fuscum... Unutar sveze 3arthecion scardici svrstane su 3 asocijacije, zajedno sa još jednom dodatnom koja gravitira ovoj svezi. Dijagnostičke vrste na nivou Srbije su: 3arthecium scardicum, Willemetia stipitata subsp. albanica, Pinguicula balcanica, Pseudorchis frivaldii, Dactylorhiza cordigera subsp. bosniaca, Carex ferruginea. Unutar sveze Caricion davallianae svrstano je 5 asocijacija, a još 5 gravitira ovoj svezi. Dijagnostičke vrste na nivou Srbije su: Eriophorum latifolium, Carex panicea, Carex paniculata, Carex hostiana, Carex distans, Juncus articulatus, Molinia caerulea, Prunella vulgaris, Valeriana dioica ssp. simplicifolia, Campylium stellatum. Ovom disertacijom iznet je novi predlog sintaksonomije tresavske vegetacije Srbije, kojim je došlo do sinonimiziranja i podređivanja sveze Carici-3ardion V. Ranđelović 1998 svezi Caricion fuscae, sveza Salici-Betulion pubescentis V. Randjelović 1994 se nije kao zasebna izdvojila, a sveza Rhynchosporion albae Koch 1926 je odbačena, dok su njene asocijacije svrstane u Caricion fuscae i Sphagno-Caricion canescentis. Na osnovu klaster analize (PC-ORD) i ordinacione DCA analize fitocenoloških snimaka tresavske vegetacije izdvojeno je 7 klastera označenih kao 1: „Sphagnum - Carex rostrata“ grupa, 2: „Braon mahovine - Carex rostrata“ grupa, 3: „3ardus stricta - Eriophorum angustifolium“ grupa, 4: „3arthecium scardicum“ grupa, 5: „Carex nigra“ grupa, 6: „Comarum palustre - Menyanthes trifoliata“ grupa i 7: „Braon mahovine - Eriophorum latifolium“ grupa. Taksonomski spektar najbrojnijih familija i rodova vaskularne flore ukazuje da je na tresavama najzastupljenija familija Cyperaceae (10,8%), zatim slede Compositae i Gramineae (9,9%), Rosaceae (5,8%), Ranunculaceae (4,7%) i druge, dok je kod rodova najzastupljeniji Carex (8,4%), zatim slede Ranunculus (3,5%), Juncus (2,9%), Trifolium (2,9%) i drugi. Analiza životnih formi vaskularne flore tresava Srbije ukazuje na apsolutnu dominaciju hemikriptofita koje čine 60,5%, slede geofite 18,6%, terofite 8,1%, fanerofite 5,2%, hamefite 4,4% i hidrofite 1,7%. Horološkom analizom utvrđeno je procentualno učešće 9 osnovnih areal tipova u flori tresava: Borealni (23%), Evroazijski (22%), Evroazijsko-planinski (21%), Srednjeevropski (21%), Holarktički (8%), Arkto-alpijski (2%), Mediteransko-submediteranski (1%), Kosmopolitski (1%) i Pontski (0,3%). U odnosu na komparativne rezultate horoloških analiza areal tipova na nivou pojedinačnih sveza i klase Scheuchzerio-Caricetea fuscae, najveća odstupanja utvrđena su kod sveze 3arthecion scardici. Konačno, ova studija sagledana u celini rezultat je pokušaja da se sistematizuju, terenski istraže, dopune i analiziraju svi dostupni podaci i saznanja o tresavskoj vegetaciji klase Scheuchzerio-Caricetea fuscae u Srbiji. U tom smislu, analiziran je položaj tresavske vegetacije Srbije unutar tresava u širem smislu, kako na nacionalnom nivou tako i na globalnom, dopunjena su saznanja o distribuciji tresavske vegetacije Srbije, njenim ekološkim osobenostima, sintaksonomskim i taksonomskim karakteristikama, trenutnom stanju i stepenu ugroženosti. U isto vreme, rezultati ove studije otvaraju i čitav niz novih pitanja i problema. S tim u vezi, predstoji dalja dopuna podataka kao i dodatne, finije analize i komparacije u cilju izvođenja odgovarajućih novih zaključaka.

Serbia and Balkan Peninsula represents the southern limits of mires distribution in Europe. Mire vegetation, or vegetation syntaxonomically included in classes Oxycocco- Sphagnetea Br.-Bl. et Tüxen ex Westhoff et al. 1946 and Scheuchzerio-Caricetea fuscae (Nordh. 1936) R. Tx. 1937. on the territory of Republic of Serbia is presented only by vegetation class Scheuchzerio-Caricetea fuscae. This vegetation is represented by a number of small and very small mountainous mires (fens) and is the main subject of this doctoral thesis. There are several aims of this thesis: shaping the general theoretical assumptions and definitions of mires and peats, their genesis, ecology, distribution and importance; systematization of all relevant published and unpublished data; analysis of existing classification systems; syntaxonomical and floristic analysis of data collected on fens in Serbia; determining the current situation and the factors threathening mires flora and vegetation; analysis of national and international legislation in the field of nature protection; analysis of protected mire areas in Serbia with perceived strengths and weaknesses of protection. Proposed revision of national syntaxonomy of fen vegetation is based on the comparative analysis of all the available vegetation plots of the class Scheuchzerio- Caricetea fuscae in Europe, including Serbia (Peterka et al. 2016 submitted). Thus, all plant associations are grouped in two orders: Caricetalia fuscae W. Koch 1926 emend. Nordhagen 1937 and Caricetalia davallianae Br.-Bl. 1950, and four alliances: Caricion fuscae Koch 1926, Sphagno-Caricion canescentis Passarge (1964) 1978, 3arthecion scardici Horvat ex Lakušić 1968 and Caricion davallianae Klika 1934. Alliance Sphagno (recurvi)-Caricion canescentis is recorded for the first time in Serbia and Caricion davallianae was previously only cited in the literature but without specific published phytocoenological plots. Newly described plant associations are only provisionally named and definitive processing awaits them for final publication. 19 associations are classified within alliance Caricion fuscae. Diagnostic species in Serbia are: Carex nigra, Eriophorum angustifolium, Carex echinata, 3ardus stricta, Sphagnum subsecundum. Alliance Sphagno-Caricion canescentis is represented by four associations. Diagnostic species in Serbia are: Carex rostrata, Equisetum fluviatile, Menyanthes trifoliata, Polytrichum commune, Sphagnum fallax, Sphagnum flexuosum, Sphagnum fuscum... Three associations are classified within the alliance 3artecion scardici as well as one additional gravitating to this alliance. Diagnostic species in Serbia are: 3arthecium scardicum, Willemetia stipitata subsp. albanica, Pinguicula balcanica, Pseudorchis frivaldii, Dactylorhiza cordigera subsp. bosniaca, Carex ferruginea. Five associations are classified within alliance Caricion davallianae, with five more being close to it. Diagnostic species are: Eriophorum latifolium, Carex panicea, Carex paniculata, Carex hostiana, Carex distans, Juncus articulatus, Molinia caerulea, Prunella vulgaris, Valeriana dioica ssp. simplicifolia, Campylium stellatum. This thesis set forth a new proposal of fen vegetation taxonomy in Serbia including following remarks: synonymy and subordination of alliance Carici-3ardion V. Ranđelović 1998 to Caricion fuscae, alliance Salici-Betulion pubescentis V. Randjelović 1994 did not step aside as separate, alliance Rhynchosporion albae Koch 1926 is rejected, while its associations are classified within Caricion fuscae and Sphagno-Caricion canescentis. Phytocoenological plots of analysed vegetation are separated in 7 clusters based on cluster analysis (PC-ORD) and a coordination DCA analysis: 1. „Sphagnum - Carex rostrata“ group, 2. „Brown mosses - Carex rostrata“ group, 3. „3ardus stricta - Eriophorum angustifolium“ group, 4. „3arthecium scardicum“ group, 5. „Carex nigra“ group, 6: „Comarum palustre - Menyanthes trifoliata“ group and 7. „Brown mosses - Eriophorum latifolium“ group. The taxonomic range of the most numerous families and genera of vascular flora indicates that the most common family is Cyperaceae (10.8%), followed by Compositae and Gramineae (9.9%), Rosaceae (5.8%), Ranunculaceae (4.7%) and others, while the most common genus is Carex (8.4%) followed by Ranunculus (3.5%), Juncus (2.9%), Trifolium (2.9%) and others. Life form analysis of vascular flora of fens from Serbia points to absolute dominance of hemicryptophytes that make 60.5%, followed by geophytes (18.6%), therophytes (8.1%), phanerophytes (5.2%), chamaephytes (4.4%) and hydrophytes (1.7%). Chorological analyses showed the percantage of 9 basic areal types in the fen flora: Boreal (23%), Eurasian (22%), Eurasian-mountain (21%), Central European (21%), Holarctic (8%), Arctic-alpine (2%), Mediterranean-submediterranean (1%), Cosmopolitan (1%) and Pontic (0.3%). In relation to comparative chorological analysis of areal type at the level of individual alliances and class Scheuchzerio-Caricetea fuscae, largest discrepancies were found in alliance 3arthecion scardici. At the end, this study viewed as a whole is the result of attempts to systematize, explore in the field, amend and analyze all the available data and knowledge about fen vegetation of class Scheuchzerio-Caricetea fuscae in Serbia. In that sense, position of the fen vegetation in Serbia within the mires in wider sense is analyzed, both at the national and global level, and data about the distribution of fens in Serbia, their ecological, syntaxonomic and taxonomic characteristics, current status and degree of threat, are discussed. At the same time, the results of this study open up a range of new issues and problems. Thus, further data gathering and additional, finer analysis and comparisons are ahead of us in order to make adequate new conclusions.

Mogućnost unapređenja standarda konzervacije, restauracije i valorizacije bojenih lazura na srebru u polihromiji baroknog perioda u srpskim pravoslavnim crkvama

The possibility of improving the standard of conservation, restoration and valorization of colored glazes on silver leaf inthe baroque polychromy of Serbian orthodox churches

Predmet ovog istraživanja je polihromna dekoracija bojenim lazurima na srebru karakteristična za period baroka u enterijeru srpskih pravoslavnih crkava. Na reprezentativnim primerima proučeni su koreni i pojava bojenih lazura na srebru, a kroz konzervatorske i fizičko-hemijske analize na uzorcima opisani materijali i tehnika izrade, kako baroknih slojeva, tako i naknadnih preslika. Analizirana je ranija konzervatorska praksa u odnosu na savremene principe u zaštiti i prezentaciji kulturnog nasleđa. Cilj istraživanja je definisanje smernica i preporuka za naučni pristup konzervaciji i restauraciji bojenih lazura na srebru kako bi se omogućila potpunija valorizacija enterijera baroknog perioda u srpskim pravoslavnim crkvama. Barokna dekoracija obuhvaćena ovim istraživanjem je retko vidljiva, zbog degradacije materijala, kasnijih preslika i konzervatorsko restauratorskih radova, što daje osiromašenu i nepotpunu predstavu baroknog enterijera. Polazeći od činjenice da su bojeni lazuri na srebru karakterističan deo polihromne dekoracije baroknih enterijera u srpskim pravoslavnim crkvama, konzervatorski pristup treba da kombinuje analizu površine, karakterizaciju materijala, istorijski, simbolički i tehnološki značaj bojenih slojeva kao i značaj samog predmeta u simboličkom i umetničkom smislu i okruženja u kome se nalazi. U praktičnoj realizaciji uzimaju se u obzir i tehnološke mogućnosti razdvajanja i prezentacije slojeva i veličina dekorisanih površina. Sagledavajući kompleksnu zavisnost materije umetničkog predmeta, iluzije koja se preko nje stvara i istorijskih vrednosti enterijera, konzervatorski pristup zasnovan na ciljanoj proceduri minimizira opasnost da se previde značajne karakteristike bojenih slojeva daje argumente za konzervaciju, restauraciju i/ili rekonstrukciju, čak i u nekim slučajevima kada su podaci o dekoraciji bojenim lazurima na srebru naizgled izgubljeni.

The subject of the research is the polychromy with colored glazes on silver leaf, characteristic of the baroque period in the interiors of Serbian orthodox churches. The origins and the appearance of colored glazes on silver leaf are studied by examining representative examples. The materials and the techniques of baroque paint layers, as well as overpaintings, are described through conservation diagnostics and physical and chemical analysis of the samples. The earlier conservation practice is studied in relation to the modern practices in the protection and presentation of cultural heritage. The aim of the research is to define guidelines and recommendations for the scientific approach to the conservation and restoration of colored glazes on silver leaf, so as to enable better valorization of the baroque interiors in Serbian orthodox churches. The baroque decoration covered in this research is rarely visible due to the material deterioration, later overpaintings and conservation treatments, leading to the poor and incomplete presentation of the baroque interiors. Starting from the fact that the colored glazes on silver leaf are a characteristic of the baroque interiors in Serbian orthodox churches, the conservation approach should combine surface analysis, material characterization, historical, symbolical and technological significance of the paint layers, as well as symbolic and artistic importance of the object and its surroundings. In the practical realization, one should also consider technological possibilities of the separation and presentation of the layers, as well as the dimension of the decorated surface. Considering the complex relation between the matter of the artistic object, the illusion achieved using this matter and the historical importance of the interior, the conservation approach based on the targeted procedure minimizes the threat to overlook important characteristics of the paint layers and offers arguments for conservation, restoration and/or reconstruction of the polychrome decoration, even in cases when data on colored glazes seems already lost.

Integrisano projektovanje konstruktivnih sistema zasnovanih na primeni ferocementa

Integrated design of structural systems based on the application of ferrocement

Predmet ovog istraživanja je analiza principa projektovanja konstrukcija zasnovanih na primeni ferocementa u okviru integrisanog pristupa projektovanju i vrednovanju ostvarenih rezultata. Predmetna analiza se vrši u skladu sa istraživačkim pitanjem: Po kojim principima treba projektovati konstrukcije zasnovane na primeni ferocementa da bi se omogućilo postizanje ekološkog kvaliteta zgrada koji je u skladu sa preporukama za održivi razvoj građenja? Osnovni cilj istraživanja je uspostavljanje teorijske osnove metodološkog koncepta integrisanog projektovanja konstrukcija, u okviru integrisanog pristupa projektovanju i vrednovanju zgrada, u funkciji ostvarenja ekološkog kvaliteta zgrada. Uži cilj istraživanja je uspostavljanje teorijsko-metodološke osnove projektovanja i vrednovanja konstruktivnih materijala u okviru integrisanog pristupa projektovanju zgrada. Istraživanje polazi od pretpostavke da integrisano projektovanje konstrukcija zasnovanih na primeni ferocementa omogućava postizanje ekološkog kvaliteta zgrada koji je u skladu sa preporukama za održivi razvoj građenja, koje se tiču „redukovanja negativnih uticaja na životnu sredinu i potrošnje resursa usled izgradnje, upotrebe i razgradnje izgrađenih kapaciteta, uz istovremen porast kvaliteta života i zdravlja i sigurnosti u izgrađenom okruženju“.1 U okviru istraživanja, ispitivanje principa integrisanog projektovanja konstrukcija u okviru integrisanog pristupa projektovanju i vrednovanju zgrada je primarno zasnovano na metodološkim principima teorije sistema.2 Reč je o metodološkom i konceptualnom aparatu za shvatanje i naučno uobličavanje problema sistema-zgrada. U prvom segmentu istraživanja ispitivan je pojam ekološkog kvaliteta zgrade. Polazište za istraživanje ovog pojma je humana ekologija. Za granicu 1 Working Group for Sustainable Construction, An agenda for sustainable construction in Europe, 11. 2 Ludwig Bertalanffy, General System theory: Foundations, Development, Application (New York: George Braziller, 1976). II sistema kome se pripisuje ekološki kvalitet je uzeta zgrada. Za vremenski okvir tokom koga se procenjuje ekološki kvalitet zgrade je uzet životni ciklus.3 Ekološki kvalitet zgrade je u okviru istraživanja definisan kao nivo do koga performanse zgrade zadovoljavaju potrebe i očekivanja njenih korisnika, a koje se odnose na socijalne i ekonomske dobrobiti ostvarene uz istovremenu zaštitu i unapređenje životne sredine tokom životnog ciklusa zgrade. Nakon kritičke analize postojećih sistema za procenu kvaliteta zgrada u kontekstu održivosti, kao i analize evropskih standarda vezanih za održivu izgradnju, u okviru istraživanja su predloženi indikatori za integralnu procenu ekološkog kvaliteta zgrade, razvrstani po međusobno povezanim i uslovljenim kriterijumima održivosti u tri grupe: indikatori u okviru kriterijuma zaštite životne sredine, indikatori u okviru kriterijuma socijalnih dobrobiti i indikatori u okviru kriterijuma ekonomskih dobrobiti. Indikatori ekološkog kvaliteta zgrada, pored toga što su osnov za integralnu procenu kvaliteta projektovanih rešenja, preko kvantitativne i kvalitativne procene nivoa projektovanih i postignutih performansi, u okviru istraživanja su uzeti kao osnov za uspostavljanje integrisanih projektnih ciljeva. Integrisanih projektni ciljevi postaju osnov za analizu principa integrisanog projektovanja konstrukcija u funkciji ostvarenja ekološkog kvaliteta zgrada. U drugom segmentu istraživanja ispitivani su principi projektovanja konstrukcija zgrada u skladu sa integrisanim projektnim ciljevima. Prvobitno je ispitivano integrisano projektovanje zgrada, zasnovano na ciljanim integrisanim performansama zgrada tokom njihovog životnog ciklusa. Ovaj vid projektovanja zgrada je definisan kao projektovanje čiji su ishod zgrade koje uspešno izvode svoju funkciju, u skladu sa svim aspektima održivosti: socijalnim, ekonomskim i aspektima zaštite životne sredine. Zatim su ispitivani principi projektovanja konstrukcije prema zahtevima kojima treba da udovolji zgrada kao celina, odnosno projektovanje konstrukcije zasnovano na ciljanim integrisanim performansama zgrade tokom njenog životnog ciklusa. Ovaj vid projektovanja konstrukcija, definisan kao integrisano projektovanje konstrukcija, zasnovan je na sistemskom pristupu, u okviru koga se konstrukcija zgrade posmatra kao funkcionalna jedinica, odnosno kao podsistem zgrade, čije je ponašanje usmereno ka cilju 3 European Committee for Standardization, EN 15978:2011. III sistema-zgrade - ekološkom kvalitetu. U ovom procesu konstrukcija se ne može razumeti, a time i vrednovati, bez razumevanja njene veze sa specifičnom funkcijom arhitektonskog prostora. Vrednuje se doprinos konstrukcije performansama zgrade čiji je sastavni deo, tokom životnog ciklusa. Reč je o procesu projektovanja u kome se koncepti vrednuju kroz agregaciju činjeničnih i vrednosnih sudova, u skladu sa integrisanim kriterijumima, odnosno kvantitativnim i kvalitativnim indikatorima ekološkog kvaliteta zgrada. Nakon spoznavanja mogućih oblika povezanosti i zavisnosti podsistema zgrade, konstrukcije i konstruktivnih materijala, odnosno ponašanja ovih podsistema usmerenih ka postizanju cilja sistema-zgrade – ekološkog kvaliteta, u okviru istraživanja su izvedeni kriterijumi za integrisano projektovanje i vrednovanje konstrukcija zasnovani na ciljanim integrisanim performansama zgrade tokom njenog životnog ciklusa, u funkciji ostvarenja ekološkog kvaliteta zgrada. Izvedeni kriterijumi tiču se smanjenja štetnih emisija u vazduh, vodu i zemljište, kao i povećanja efikasnosti korišćenja resursa, odnosno smanjenja intenziteta njihovog korišćenja, kao i realizacije arhitektonskih prostora koji doprinose kvalitetu života ljudi, odnosno životnom zadovoljstvu, kroz očuvanje i unapređenje njihovog zdravlja, podsticanje osećaja sigurnosti, ugodnosti i sklada sa životnim okruženjem. U trećem segmentu istraživanja ispitivani su principi projektovanja ferocementa i konstrukcija zasnovanih na njegovoj primeni, u skladu sa kriterijumima za integrisano projektovanje i vrednovanje konstrukcija zgrada, kao i ekološki efekti primene ovog kompozitnog materijala u sklopu konstrukcija, prema kriterijumu zaštite životne sredine tokom životnog ciklusa zgrade i prema kriterijumu socijalnih dobrobiti tokom faze upotrebe zgrade. Ferocement, kao kompozitni materijal, je razmatran kao podsistem konstrukcije zgrade. Njegovo projektovanje je analizirano kroz odnose i veze sa podsistemom višeg reda- konstrukcijom zgrade i sistemom-zgradom, kao i kroz njegovo ponašanje usmereno ka postizanju cilja sistema-zgrade - ekološkog kvaliteta. Istraživanjem su produbljeni i prošireni projektantski „alati za definisanje svojstava i korišćenje ferocementa“.4 Stvorena je saznajna osnova projektovanja i vrednovanja 4 Komisija za standarde iz oblasti Betonskih konstrukcija, Tehnička preporuka za ferocement, 2. IV ferocementa i konstrukcija zasnovanih na njegovoj primeni, u okviru integrisanog pristupa projektovanju zgrada. Istraživanjem je proverena i potvrđena polazna hipoteza po kojoj integrisano projektovanje konstrukcija zasnovanih na primeni ferocementa omogućava postizanje ekološkog kvaliteta zgrada koji je u skladu sa preporukama za održivi razvoj građenja. Istraživanje je pokazalo da novi “uslovi” projektovanja zgrada, koji prema Zakonu o planiranju i izgradnji Republike Srbije treba da „obezbede održivu gradnju“ ("Sl. glasnik RS", br. 72/2009, 81/2009 - ispr., 64/2010 - odluka US, 24/2011, 121/2012, 42/2013 - odluka US i 50/2013 - odluka US, član 2), pre svega treba da se zasnivaju na integraciji projektnih ciljeva, kao i na integralnoj proceni kvaliteta projektovanih rešenja tokom životnog ciklusa, u skladu sa kvantitativnim i kvalitativnim indikatorima ekološkog kvaliteta zgrada. Projektovanje ekološki prihvatljivih rešenja treba da bude zasnovano na strategijama istraživanja koje prelaze granice disciplina, kako bi se mogli spoznati odnosi i veze između podsistema zgrade, konstrukcije i konstruktivnih materijala, kao i kako bi moglo biti spoznato ponašanje ovih podsistema usmereno ka postizanju cilja sistema-zgrade - ekološkog kvaliteta. U pitanju je sistemski pristup koji za svoj rezultat u praktičnoj operacionalizaciji ima koncipiranje sistema-zgrada različitih odnosa i veza između podsistema, konstrukcije i konstruktivnih materijala, odnosno koncipiranje zgrada različitih kvantitativnih i kvalitativnih svojstva, sa jednim zajedničkim višim svojstvom – ekološkim kvalitetom.

The subject of this research is to analyse the principles of structural design based on the application of ferrocement within an integrated approach to the design, and evaluation of the achieved results. The analysis concerned is performed according to the research question: What principles should be applied to design the structures based on the application of ferrocement, to enable the achievement of ecological quality of buildings, in line with recommendations for sustainable building development? The main goal of this research is to establish the theoretical basis of the methodological concept of integrated structural design, within an integrated approach to the design and evaluation of buildings for the purpose of achieving ecological quality of buildings. The specific objective of the research is to establish a theoretical and methodological basis of the design and evaluation of construction materials in the framework of an integrated approach to building design. The study assumes that the integrated structural design based on the application of ferrocement enables achieving ecological quality of buildings, which is in line with recommendations for sustainable building development, related to the "reduction of negative environmental impact and resource consumption due to construction, use and dismantling of constructed facilities, with a simultaneous increase in life quality of and health and safety in the built environment“.5 Examining the principles of integrated structural design within an integrated approach to the design and evaluation of buildings, in this research is primarily based on the methodological principles of the system theory.6 This is the methodological and conceptual apparatus for understanding and scientific shaping of the problem of system–buildings. The first segment of the research examined the concept of the ecological quality of a building. The starting point to explore this concept is the human ecology. A building is taken as a boundary of system which the ecological quality is attributed to. A life-cycle is 5 Working Group for Sustainable Construction, An agenda for sustainable construction in Europe, 11. 6 Ludwig Bertalanffy, General System theory: Foundations, Development, Application (New York: George Braziller, 1976). VI taken as the time frame during which the ecological quality of the building is assessed.7 The ecological quality of the building within this research was defined as the extent to which performances of the building meet the needs and expectations of its users, which are related to the social and economic benefits achieved with simultaneous protection and improvement of the environment throughout the life-cycle of the building. After a critical analysis of the existing systems to assess the quality of buildings in the context of sustainability, as well as analysis of European Standards for Sustainable Construction, indicators for the integral assessment of ecological quality of buildings were proposed within the research, classified by the interrelated and conditioned sustainability criteria into three groups: indicators within the environmental criteria, indicators within the criteria of social well-being, and indicators within the criteria of economic well-being. Indicators of ecological quality of building, in addition to being the basis for an integrated assessment of the quality of the designed solution, through quantitative and qualitative assessment of the level of designed and achieved performances, within the research were taken as the basis for establishment of integrated project objectives. Integrated project objectives have become the basis for the analysis of integrated structural design principles for the purpose of achieving ecological quality of buildings. In the second segment of the research, the principles of design of building structures were examined in accordance with the integrated project objectives. Originally, the integrated design of buildings was studied, based on targeted integrated performances of buildings during their life-cycle. This form of building design is defined as a design which results in buildings that successfully carry out their function, in accordance with all aspects of sustainability: social, economic and environmental aspects. Then the principles of structural design were examined according to requirements to be satisfied by a building as a whole, that is, structural design based on targeted integrated performances of a building throughout its life-cycle. This type of structural design, defined as the integrated structural design, is based on a systemic approach, in which the structure of the building is seen as a functional unit, i.e., as a sub-system of the building, whose behavior is directed towards the aim of system-building – ecological quality. In this process, a structure cannot be understood, and thus evaluated, without understanding its relationship with the specific function of architectural space. The evaluation is made of the contribution of a structure to 7 European Committee for Standardization, EN 15978:2011. VII performances of the building which is its integral part, throughout the life-cycle. It is a design process in which concepts are evaluated through aggregation of fact and value judgments in accordance with the integrated criteria, that is, quantitative and qualitative indicators of ecological quality of buildings. After comprehending the possible forms of connection and dependency of sub-systems of a building, building structure and construction materials, as well as the behavior of these sub-systems directed towards achieving the goal of the system-building – ecological quality, criteria for the integrated design and assessment of structures based on targeted integrated performances of a building during its life-cycle were derived within the research, for the purpose of achieving ecological quality of buildings. The derived criteria are related to the reduction of harmful emissions into the air, water and soil, as well as to the increase of the resources use efficiency, i.e., reduction of the intensity of their use, as well as to the realization of architectural spaces that contribute to the quality of people's lives and life satisfaction, through preservation and improvement of their health, incitement of the sense of security, amenity and harmony with the environment. In the third segment of the research, the principles of ferrocement design and structures based on its application were studied, in accordance with the criteria for integrated design and assessment of building structures, as well as the ecological effects of the application of this composite material within structures, in accordance with the criteria for the environmental protection during the life cycle of building and in accordance with the criteria for the social well-being during the use phase of a building. Ferrocement, as a composite material, was considered as a subsystem of the building structure. Its design was analyzed through relations and links with the subsystem of a higher order – structure of the building and system-building, as well as through its behavior directed towards achieving the aim of system-building – ecological quality. With this one research, the designers "tools for defining the properties and use of ferrocement"8 were further developed and expanded. A cognitive basis of the design and evaluation of ferrocement and structures based on its application was created, within the integrated approach to the design of buildings. The research tested and confirmed the initial hypothesis according to which the integrated design of structural systems based on the application of ferrocement enables achieving ecological quality of buildings, which is in line with recommendations for 8 Komisija za standarde iz oblasti Betonskih konstrukcija, Tehnička preporuka za ferocement, 2. VIII sustainable building development. The research has shown that new "conditions" of building design, which should "ensure sustainable building" according to the Law on Planning and Construction of the Republic of Serbia ("Official Gazette of the Republic of Serbia", no. 72/2009, 81/2009 - rev., 64/2010 – Constitutional Court Decision, 24/2011, 121/2012, 42/2013 – Constitutional Court Decision and 50/2013 – Constitutional Court Decision, clause 2), above all, should be based on the integration of project objectives, as well as integral quality assessment of the designed solutions throughout the life-cycle, in accordance with the quantitative and qualitative indicators of ecological quality of buildings. Designing ecologically acceptable solutions should be based on research strategies that transcend disciplinary boundaries, in order to comprehend the relations and connections between the sub-systems of the building, building structure and construction materials, as well as the behavior of these sub-systems directed towards achieving the goal of the system-building – ecological quality. This is a systemic approach whose result, in its practical operationalization, is conception of system-buildings of different relationships and connections between sub-systems, building structure and construction materials, that is, conception of buildings of various quantitative and qualitative properties, with a higher common property – ecological quality.

Konstruktivno-geometrijsko generisanje kupola sa konkavnim poliedarskim površima

Constructive - geometric generating of cupolae with concave polyhedral surfaces

Konkavne kupole su poliedri koji slede način generisanja kupola (Džonsonovih tela), s tim što izostaje kriterijum konveksnosti i što se u omotaču ovih tela pojavljuju dva ili više niza jednakostraničnih trouglova.

Concave Cupolae are polyhedra which follow the method of generating geometric cupolae (Johnson´s solids), except for the convexity criterion which is omitted, and the fact that in the lateral surface of the solids appear two or more series of equilateral triangles

Optimizacija stepena transparentnog zastaklenja fasade administrativnih zgrada u klimatskim uslovima Beograda

Optimisation of transparent ratio in highly glazed office building facades for Belgrade climate conditions.

U savremenoj poslovnoj arhitekturi koncept materijalne transparentnosti se eksploatiše u velikom obimu, kako bi se, preko pojavnosti transparentne arhitekture, izrazila moć i pristupačnost preduzeća. Velike površine transparentnog stakla na fasadi zgrade, predstavljaju reklamu institucije ili preduzeća, putem koje se potencijalnim klijentima prenosi poruka o njihovoj pristupačnosti, otvorenosti i poštenju u poslovanju. Primenjena visoka tehnologija u izradi staklenih površina fasade, takođe asocira na imidž moderne, savremene organizacije ili institucije, koja se, pored svega ostalog, odgovorno ponaša prema očuvanju životne sredine i energije. Sa energetskog aspekta, sa primenom transparentnog zastakljenja fasade dovodi se u pitanje upravo ono što staklena fasada reklamira - odgovornost prema očuvanju životne sredine i energije, iz razloga što, veoma često, transparentni delovi fasade nisu optimizovani prema prostornim i klimatskim karakteristikama lokacije. Ovo istraživanje se bavi analizom koliko je u administrativnim prostorima u klimatskim uslovima Beograda, moguće iskorišćenje prednosti koje obezbeđuju velike staklene površine: pre svega iskorišćenje dnevne svetlosti, kako bi se smanjile potrebe zgrade za električnim osvetljenjem i iskorišćenje solarnih toplotnih dobitaka, kojima bi se kompenzovali veliki transmisioni gubici u zimskom periodu. Istraživanje je sprovedeno primarno sa aspekta održanja vizuelnog i svetlosnog komfora i integrisano sa istraživanjem energetskih performansi, kao posledice primenjenih mera za održanje vizuelnog komfora u administrativnim prostorima. Istraživanja svetlosnih i vizuelnih performansi prostora putem kompjuterske simulacije su veoma aktuelna, budući da sve do pre deset godina nije bila razvijena odgovarajući precizna tehnologija simulacije, niti parametri za izražavanje svetlosnih performansi prostora. U ovom istraživanju kompjuterska simulacija performansi prostora izvedena je korišćenjem programa DIVA for Rhino, koji je razvijen na Harvard univerzitetu i prvi put predstavljen 2010. godine. Prednost programa je integrativno sagledavanje svetlosnih i vizuelnih performansi prostora sa energetskim aspektima prostora. Istraživanje je sprovedeno kompjuterskom simulacijom performansi za modele individualne kancelarije sa stepenom zastakljenja od 50%, 60%, 70% i maksimalnih 85%, za četiri grupe stakla sa svetlosnom propustljivosti od 50% do 80% i prema četiri osnovne orijentacije. Analizirani su prostori bez integrisanih zastora kako bi se prvo utvrdili potencijali iskorišćenja dnevne svetlosti i energije za različite kombinacije varijabilnih parametara. Osnovni predmet istraživanja su administrativni prostori kod kojih su integrisani zastori u sistemu zastakljenja, budući da u administrativnim prostorima uspostavljanje vizuelnog i svetlosnog komfora zavisi od upotrebe zastora. Rezultati pokazuju da se u klimatskim uslovima Beograda visok stepen zastakljenja fasade može primeniti na fasadama koje su orijentisane prema jugu i prema severu. Pri severno orijentisanim prostorima ostvaruje se najveće iskorišćenje dnevne svetlosti. i najmanje potrebe za energijom za osvetljenje, ali su ukupne potrebe za energijom slične kao kod prostora koji su istočno i zapadno orijentisani i sa nižim stepenom zastakljenja. Iskorišćenje dnevne svetlosti kod severno orijentisanih prostora je najoptimalnije sa primenom stakla sa visokom svetlosnom i energetskom propustljivosti zračenja u kombinaciji sa izuzetno dobrim izolacionim karakteristikama. Tokom perioda zagrevanja severna fasada nije izložena solarnom zračenju, nema značajnih solarnih toplotnih dobitaka, tako da se sa dobrim izolacionim karakteristikama i energetskom propustljivosti zračenja obezbeđuje kontrola izuzetno visokih energetskih potreba za zagrevanje. U isto vreme, kako prostor nije izložen direktnom zračenju, zastori se spuštanju veoma kratko tokom godine, što obezbeđuje visok nivo iskorišćenja dominantno difuzne svetlosti. Kod južno orijentisanih prostora iskorišćenje dnevne svetlosti je relativno manje u odnosu na sve ostale orijentacije zbog izloženosti solarnom zračenju tokom ukupnog perioda korišćenja prostora, koje dovodi do veoma čestog spuštanja zastora. Sa povećanjem stepena zastakljenja fasade, ostvaruju se uštede u energetskim potrebama za osvetljenje, kao i pri svim drugim orijentacijama, ali su uštede relativno male. Najveće iskorišćenje se ostvaruje sa primenom stakla sa malom svetlosnom propustljivosti, koja prouzrokuju duži period podignutih zastora, a time i efikasnije iskorišćenje dnevne svetlosti. Primarni razlog zbog kojeg je energetski efikasna primena visokog stepena zastakljenja kod južno orijentisanih prostora je visok nivo solarnih toplotnih dobitaka u periodu zagrevanja, koji obezbeđuje skoro dvostruko manje energetske potrebe prostora za zagrevanje i oko 30% manje ukupne energetske potrebe prostora u odnosu na sve druge orijentacije.

The concept of transparency is highly exploited in contemporary office architecture, in order to express the character of company as powerful, but very accessible to clients. Highly glazed facades are used to express the openness and business honesty. High technology glazed facade is also associated with an image of modern organisation behind transparent walls, which is also responsible to the environment and energy preservation. Energy preservation responsibility becomes quite questionable if glazed facade ratio is not optimised according to building site and climate conditions. This research is exploring the utilisation of glazed facades benefits in office spaces in Belgrade climate. Such benefits are daylight utilisation (in order to reduce energy use for lighting) and utilisation of solar heat gains that compensate the large transmission heat loses during heating period. The primary aspect of this research is visual comfort in office spaces under daylight conditions, integrated with its energy implications. The research of daylight performance in office spaces using computer simulation is vary actual subject, as the technology and methodology of simulation process, as well as climate based daylight metrics, have been rapidly developed only during past decade. DIVA for Rhino was used as a simulation tool for daylight and energy performances of office spaces. The program was developed at Harvard University and presented in 2010. The advantage of DIVA program is that it has an integrated approach to daylight and energy simulations. An individual office space was analysed with net glazed ratio (WWR) of 50%, 60%, 70% and maximum possible 85%. The space was rotated to face four major orientations (east, south, west and north). Four groups of glass characteristics were selected, which correspond to 50%, 60%, 70% and 80% glass light transmittance. First part of analysis was focused on simulation of daylight and energy performances in office spaces without any shading. The goal of first simulation process was to explore only daylighting and energy use potentials in office spaces, as visual and thermal comfort conditions cannot be fulfilled in spaces without shading devices. The main subject of this research are office spaces with integrated shading devices, which are flexible to changeable climate conditions and, as such, can create comfortable working environment. The results of the analysis show that, in Belgrade climate, highly glazed facades can be used in south and north orientated office spaces. North orientated spaces have highest utilisation of daylight and consequently the lowest lighting energy use. The best optimum solution for utilisation of daylight is glass variant with high light and energy transmission characteristics combined with extremely good isolation properties. During heating period, spaces facing north are not receiving any solar gains, so high energy transmittance and isolation prosperities control the extremely high heating energy needs. At the same time, as the space is not exposed to direct solar radiation, the shading is open most time of the year, which results high utilisation of diffuse daylight. South facing spaces have high lighting energy needs. High utilisation of daylight is blocked by presence of direct radiation on facade during most time of the year, so shading is mostly closed. The same as for other orientations, lighting energy savings are raising as glazing ratio is rising, but the savings are relatively low, comparing to other energy needs. The highest daylight utilisation is occurring with low light transmission glass (low as 50%), because glass is reducing illuminance levels on working plane, so the shading stays open more often. But the primary reason for highly glazed south facade energy efficiency is high level of solar gains during heating period. Solar gains on south facade are cutting energy use for heating at a level that is almost twice lower, compared to other orientations of space, and about 30% lower overall energy.

Uticaj etanolnog ekstrakta lista masline (Olea europaea L.) na genomsku nestabilnost, parametre oksidativnog stresa i inflamacije kod pacijenata sa reumatoidnim artritisom

Effects of ethanolic olive leaf extract (Olea europaea L.) on genomic instability, parameters of oxidative stress and inflammation in rheumatoid arthritis patients

Reumatoidni artritis (RA) predstavlja autoimunsko inflamatorno oboljenje, u čijem razvoju ključnu ulogu imaju inflamatorni medijatori i oksidativni stres, koji mogu dovesti do oštećenja biomolekula i nastanka genomske nestabilnosti. Suvi etanolni ekstrakt lista masline (engl. dry olive leaf extract, DOLE) je poznat po izuzetnim antioksidativnim, antiinflamatornim i genoprotektivnim dejstvima. Međutim, do sada nisu ispitivani njegovi efekti u terapiji RA. Cilj istraživanja ove doktorske disertacije je poređenje efekata tronedeljne i šestonedeljne kombinovane terapije metotreksatom i DOLE u odnosu na efekte standardne terapije samo metotreksatom, kod pacijenata sa RA. U istraživanju je učestvovalo 8 pacijenata novodijagnostikovanih za RA, 16 pacijenata sa dugotrajnim RA, koji su bili na kombinovanoj terapiji, dok je posebnu grupu činilo 8 novodijagnostikovanih pacijenata na terapiji samo metotreksatom. Kod sve tri grupe, pre uvođenja eksperimentalne terapije, zabeležen je povišen nivo inflamacije, oksidativnih oštećenja lipida, proteina i DNK, kao i povećana učestalost mikronukleusa u odnosu na vrednosti kod zdravih kontrolnih ispitanika. Rezultati tronedeljne i šestonedeljne terapije su pokazali da kombinovana primena DOLE i metotreksata daje bolje efekte u odnosu na terapiju samo metotreksatom, pre svega u pogledu efikasnije redukcije primarnih DNK oštećenja, kao i redukcije nivoa interleukina-6, kod pacijenata u ranoj fazi RA. Kombinovana terapija je takođe pokazala efekat u redukciji nitrita kod grupe u ranoj fazi RA i smanjenje lipidne peroksidacije kod dugotrajno obolelih pacijenata. Praćenjem učestalosti mikronukleusa u limfocitima, nisu utvrđeni značajni efekti terapije na nivo genomske nestabilnosti posle šest nedelja. Pokazani su pozitivni efekti kombinovane terapijske primene metotreksata i DOLE na oksidativna oštećenja ćelija kod pacijenata u ranoj fazi RA, u poređenju sa terapijom samo metotreksatom. Kombinovana terapija je bila efikasnija u ranoj fazi RA, dok su njeni efekti slabije izraženi kod dugotrajnog stanja aktivne bolesti.

Rheumatoid arthritis (RA) is an autoimmune inflammatory disease and in its development the key features are inflammatory mediators and free radical production, that may result with damage to biomolecules and cause genomic instability. Dry olive leaf extract (DOLE) is known for its exceptional antioxidant, anti-inflammatory and genoprotective effects. However, there are still no data on its effects in the treatment of RA. The aim of this doctoral thesis was to compare the effects of a three-week and six- week combined treatment with methotrexate and DOLE in comparison with the effects of standard therapy with methotrexate alone, in patients with RA. The study included 8 newly diagnosed RA patients and 16 patients with long-term RA that received concomitant treatment with DOLE and methotrexate, while another group of 8 newly diagnosed RA patients were treated only with methotrexate. The initial measurements before the introduction of the experimental treatment showed elevated levels of inflammation, oxidative damage to lipids, proteins and DNA, as well as the increased frequencies of micronuclei in all three groups of RA patients compared to healthy controls. The results of three-week and six-week therapy showed that combined use of methotrexate and DOLE achieved improved effects compared to the methotrexate therapy alone, i.e. in terms of a more rapid reduction of primary DNA damage, as well as by reducing the level of proinflammatory cytokine interleukine 6 (IL-6) in early phase RA patients. Concomitant treatment also showed an effect on reduction of nitrites in patients with early phase RA, and decreasing lipid peroxidation in long-term RA patients. The monitored frequencies of micronuclei in lymphocytes of RA patients, showed no significant effects of treatment on genomic instability after six weeks. The present study demonstrated beneficial effects of concomitant treatment with methotrexate and DOLE on modulation of oxidative cell damage in early phase RA patients with respect to the effects of standard therapy, methotrexate alone. Combined therapy was more effective in patients with newly-diagnosed RA, while the effects of treatment were less pronounced in long- term active disease.

Analiza promotorskih varijanti genaCYP2C19 i njihova uloga u predviđanju terapijskog odgovora na lek klopidogrel kod pacijenata sa infarktom miokarda i stenozom kariotida

Analysis of variants in the promoter region of CYP2C19 gene and their role in prediction of clopidogrel response in patients with myocardial infarcition and carotid stenosis

Klopidogrel je antiagregacioni lek koji je sa aspirinom, u sklopu dvojne antiagregacione terapije, indikovan u stanjima koja nose rizik od ishemijskih događaja i tromboembolijskih komplikacija. Postoji velika interindividualna varijabilnost u odgovoru na klopidogrel. Značajan broj pacijenata kao posledicu neadekvatnog odgovora na terapiju može iskusiti različita neželjena dejstva. Kod oko 9% pacijenata koji su na terapiji klopidogrelom javlja se krvarenje kao neželjeni efekat. Ono samo po sebi može biti životno ugrožavajuće, a sekundarno može biti povezano sa pojavom ishemijskih događaja. Uzroci varijabilnosti u odgovoru na terapiju klopidogrelom, mogu biti kako stečeni, tako i posledica genetičkih faktora. Među genetičkim faktorima najznačajniji doprinos ima gen za enzim CYP2C19 koji je neophodan u procesu konverzije klopidogrela u aktivnu formu leka. Zbog toga su varijante CYP2C19 gena najčešće razmatrane u okviru farmakogenetike klopidogrela. Od posebnog kliničkog značaja su varijante rs4244285 (c.681G>A; CYP2C19*2) i rs4986893 (c.636G>A; CYP2C19*3), koje su povezane sa oslabljenim odgovorom na klopidogrel. Promotorska varijanta rs12248560 (c.-806C>T, CYP2C19*17) se dovodi u vezu sa pojačanim stepenom inhibicije trombocita i povišenim rizikom od krvarenja kod pacijenata na terapiji klopidogrelom. U fokusu ove studije bilo je ispitivanje uticaja CYP2C19 gena na pojavu pojačanog odgovora na klopidogrel koje vodi povišenom riziku od krvarenja. Istraživanje je sprovedeno u grupi pacijenata koji su pretrpeli infarkt miokarda i bili podvrgnuti perkutanoj koronarnoj intervenciji (PCI), kao i grupi pacijenata sa stenozom karotida koji su podvrgnuti karotidnoj endarterektomiji (CEA). U studiji su analizirane promotorske varijante rs12248560 (c.-806C>T, CYP2C19*17) i rs11568732 (c.-889T>G, CYP2C19*20) za koje su literaturni podaci i naši preliminarni rezultati pokazali da mogu biti povezane sa pojačanim odgovorom na klopidogrel. Dodatno su analizirane egzonske varijante rs4244285 (c.681G>A; CYP2C19*2) i rs4986893 (c.636G>A, CYP2C19*3) zbog mogućeg uticaja na efekte promotorskih varijanti. Zbog oskudnosti podataka koji se odnose na promotorsku varijantu rs11568732 (c.-889T>G, CYP2C19*20), jedan od ciljeva studije bio je da se u in vitro uslovima ispita njen funkcijski značaj. Takođe, jedan od ciljeva studije bio je da se analiza varijanti gena CYP2C19, koje su pokazale značaj u pogledu terapije klopidogrelom iskoristi u svrhu racionalizacije postojećeg protokola za primenu antiagregacione terapije u Srbiji. Analiza varijante rs12248560 (c.-806C>T, CYP2C19*17) nije pokazala povezanost sa krvarenjem kod pacijenata podvrgnutih PCI nakon infarkta miokarda, koji su bili na terapiji klopidogrelom. Takođe, ova varijanta nije bila povezana sa agregacijom trombocita indukovanom adenozin difosfatom (ADP) kod pacijenata sa stenozom karotide koji su nakon karotidne endarterektomije bili na terapiji klopidogrelom. Varijanta rs12248560 (c.-806C>T, CYP2C19*17) nije bila povezana ni sa pojačanim odgovorom na klopidogrel definisanim na osnovu različitih graničnih vrednosti ADP-indukovane agregacije. Varijanta rs11568732 (c.-889T>G, CYP2C19*20) je pokazala značajnu povezanost sa krvarenjem kod pacijenata sa akutnim infarktom miokarda podvrgnutih PCI. Prisustvo alela G povećava mogućnost krvarenja 3,7 puta. Međutim, varijanta rs11568732 (c.-889T>G, CYP2C19*20) nije bila povezana sa ADP-indukovanom agregacijom kod pacijenata koji su nakon karotidne endarterektomije bili na terapiji klopidogrelom. Takođe, ova varijanta nije pokazala značajnu povezanost sa pojačanim odgovorom na klopidogrel procenjenim na osnovu različitih graničnih vrednosti ADP-indukovane agregacije. Sekvenciranjem egzona 4 gena CYP2C19 pokazano je da se alal G varijante rs11568732(c.-889T>G, CYP2C19*20) u populaciji Srbije javlja potpuno nezavisno od alela A varijante rs4986893 (c.636G>A, CYP2C19*3) sa kojim je svrstan u haplotip CYP2C19*3B. Funkcijska analiza varijante rs11568732 (c.-889T>G, CYP2C19*20) pokazala je da prisustvo nukleotida G dovodi do smanjenja aktivnosti promotora za 20%. Takođe, pokazano je da progesteronski receptor utiče na povećanje promotorske aktivnosti gena CYP2C19 nezavisno od varijante rs11568732. Glukokortikoidni receptor po aktivaciji deksametazonom dovodi do manjeg povećanja aktivnosti promotora ako se na poziciji c.-889, nalazi nukleotid G, nego wt nukleotid T. Analiza farmakogenetički vođene terapije klopidogrelom u odnosu na genotip varijante rs4244285 (c.681G>A; CYP2C19*2) pokazala se kao strategija uštede u Srbiji. Rezultati ove studije pokazuju da varijanta rs12248560 (c.-806C>T, CYP2C19*17) ne predstavlja pouzdan prognostički faktor za procenu rizika od krvarenja. Što se tiče varijante rs11568732 (c.-889T>G, CYP2C19*20) neophodna su dalja istraživanja da bi se rasvetlio njen mehanizam delovanja i pouzdano utvrdila njena uloga u krvarenju kod PCI pacijenata na terapiji klopidogrelom.

Wide inter-individual variability in clopidogrel response has been reported. Significant number of patients with inadequate response to clopidogrel experience different adverse effects. Bleeding as an adverse event occurs in about 9% of patients on clopidogrel therapy. It can be life threatening and secondarily, related to higher risk of ischemic events. Variability in clopidogrel response may be explained by different non-genetic and genetic factors. Among genetic factors, the most important role has CYP2C19 gene encoding enzyme responsible for conversion of clopidogrel to its active form. Hence, CYP2C19 gene variants are the most commonly investigated genetic variants related to clopidogrel pharmacogenetics. Clinically, the most relevant variants are rs4244285 (c.681G>A; CYP2C19*2) and rs4986893 (c.636G>A; CYP2C19*3) which lead to clopidogrel poor response. Additionally, promoter variant rs12248560 (c.-806C>T, CYP2C19*17) is associated with elevated platelet inhibition and increased risk of bleeding in patients on clopidogrel. This study was focused on investigating role of CYP2C19 gene in clopidogrel hyper- responsiveness which increases the risk of bleeding. This study was conducted in a group of patients with acute myocardial infarction who underwent percutaneous coronary intervention (PCI) as well as in patients with carotid stenosis who underwent carotid endarterectomy (CEA). In this study we examined two promoter rs12248560 (CYP2C19*17) and rs11568732 (CYP2C19*20) variants, for which literature and our preliminary results showed to be associated with elevated clopidogrel response. Additionally, we analyzed exonic variants rs4244285 (c.681G>A; CYP2C19*2) and rs4986893 (c.636G>A, CYP2C19*3) considering their potential influence on the promoter variants’ effects. Since literature data on rs11568732 (c.-889T>G, CYP2C19*20) variant is scarce, we aimed to investigate its functional significance. Further, one of our aims was the anlysis of the relevant CYP2C19 variants regarding clopidogrel therapy, which can be used to rationalize current protocols for antiplatelet therapy in Serbia. Analysis of rs12248560 (c.-806C>T, CYP2C19*17) variant showed no association with bleeding events in patients who underwent PCI after myocardial infarction and were on clopidogrel therapy. Next, we showed that this variant is not associated with ADP induced platelet aggregation in patients with carotid stenosis who underwent CEA. Additionally, rs12248560 (c.-806C>T, CYP2C19*17) variant was not associated with clopidogrel hyperresponse defined by three different cut-offs based on ADP induced platelet aggregation. Our results showed that rs11568732 (c.-889T>G, CYP2C19*20) variant is asssociated with bleeding events in patients with acute myocardial infarction who underwent PCI. We found that carriers of allele G of this variant are 3,7 times more likely to experience bleeding. However, rs11568732 (c.-889T>G, CYP2C19*20) variant was not associated with ADP induced platelet aggregation in patients with carotid stenosis who underwent CEA, nor with clopidogrel hyperresponse defined by three different cut-offs based on ADP induced platelet aggregation. Furthermore, we showed that in Serbian patients allele G of rs11568732 variant appears independently of rs4986893 (c.636G>A, CYP2C19*3) variant as well as of CYP2C19*3B haplotype. The functional analysis of rs11568732 (c.-889T>G, CYP2C19*20) variant showed that the presence of nucleotid G decreases promoter activity by 20%. Also, we showed that progesterone receptor increases CYP2C19 promoter activity independently of rs11568732 (c.- 889T>G, CYP2C19*20) variant. The glucocorticoid receptor activated by dexamethasone leads to smaller increase of promoter activity if nucleotid G is present at the position c.-889 instead of wt nucleotide T. The analysis of pharmacogenetic-guided clopidogrel therapy based on rs4244285 (c.681G>A; CYP2C19*2) genotype showed that this would be a cost-saving approach under current conditions in Serbia. Finally, results of this study show that rs12248560 (c.-806C>T, CYP2C19*17) variant is not a reliable prognostic marker for estimation of bleeding risk. Regarding rs11568732 (c.- 889T>G, CYP2C19*20) variant further investigation is needed in order to elucidate mechanism of action and to validate its effect on bleeding in PCI patients on clopidogrel treatment.

Efekat eksperimentalnog dijabetesa tipa 1 i supstitucione terapije na molekule regulisane insulinom u srcu pacova

The effect of experimentally induced Type 1 diabetes and insulin replacement therapy on insulin signalling pathways in the rat heart

Insulinska deficijencija i hiperglikemija, koje predstavljaju osnovne karakteristike dijabetesa tipa 1, povezane su sa brojnim endokrinim i metaboličkim promenama, a jedna od najčešćih komplikacija je povećana učestalost nastanka kardiovaskularnih oboljenja. Zbog toga je rana i adekvatna primena terapije insulinom neophodna, kako za kontrolu bolesti, tako i za prevenciju potencijalnih komplikacija. Polazna hipoteza istraţivanja čiji su rezultati obuhvaćeni ovom doktorskom disertacijom je bila da u stanju hipoinsulinemije i hiperglikemije moţe doći do promena u signalnom putu insulina u srcu, što moţe da se odrazi na produkciju azot-monoksida i iskorišćavanje energetskih supstrata, a da bi supstitucija insulinom trebala da koriguje nastale promene. Za uspostavljanje eksperimentalnog modela dijabetesa tipa 1, muţjaci pacova soja Wistar tretirani su streptozotocinom (60 mg/kg). Poznato je da se akutni dijabetes javlja između osmog dana i tri nedelje, a hronični 3 nedelje nakon tretmana streptozotocinom. Ţivotinje sa dijabetesom su ili bile netretirane 2 nedelje ili podvrgnute subhroničnom tretmanu insulinom (3 IU) u trajanju od 7 dana. Za analizu su selektirani signalni molekuli Akt i ERK koji predstavljaju medijatore gotovo svih efekata insulina u srcu. Osim toga analizirani su i efektorni molekuli regulisani preko aktivacije signalnih puteva Akt i ERK, kao što su eNOS i iNOS, uključeni u sintezu azot-monoksida, kao i transporteri glukoze GLUT1 i GLUT4 i transporter masnih kiselina CD36. Ispitivana je ekspresija ovih molekula na nivou gena i proteina i/ili fosforilacija, kao i unutarćelijska lokalizacija transportera za glukozu i masne kiseline u srcu. Poznato je da je kompeticija između arginaze i NOS za zajednički supstrat L-arginin, ograničavajući faktor za proizvodnju azot-monoksida. U akutnoj fazi streptozotocinom izazvanog dijabetesa u srcu pacova zapaţeno je povećanje aktivnosti enzima arginaze što je praćeno smanjenjem koncentracije L-arginina. Supstitucija insulinom je smanjila aktivnost arginaze i povećala koncentraciju L-arginina, što ukazuje na značajan regulatorni efekat insulina na aktivnost arginaze. Tretman streptozotocinom je doveo do smanjenja fosforilacije eNOS na Ser1177, i povećanja nivoa iRNK i proteina iNOS u srcu pacova. Supstitucija insulinom je vratila nivo fosforilacije eNOS i nivo proteina iNOS na kontrolni. Iz ovih rezultata proizilazi da je u stanju dijabetesa pomerena ravnoteţa ka korišćenju L-arginina od strane arginaze, a da je tretman insulinom uspostavio normalnu aktivnost NOS. Kod pacova sa dijabetesom došlo je do smanjenja nivoa iRNK za Akt i fosforilacije na Ser473, kao i fosforilacije ERK1/2 na Thr202 i Tyr204. Supstitucija insulinom je vratila nivo iRNK za Akt i fosforilaciju na nivo kontrole, međutim nije imala efekta na kinaznu aktivnost ERK1/2. Moţe se zaključiti da u stanju dijabetesa dolazi do promena u oba signalna puta, a da je tretman insulinom bio uspešan samo u ispravljanju promena u PI3K/Akt putu. Insulin je uključen u regulaciju metabolizma ugljenih hidrata i lipida u srcu. U dijabetesu dolazi do metaboličkih odstupanja u korišćenju energetskih supstrata, što rezultira u smanjenom preuzimanju glukoze i povećanom korišćenju masnih kiselina. Ekspresija gena i nivo proteina GLUT4 i GLUT1 su bili smanjeni u srcu pacova sa dijabetesom, a tretman insulinom je uglavnom vratio njihovu ekspresiju na nivo kontrole. Tretman streptozotocinom je smanjio nivo proteina GLUT4 i GLUT1 u frakciji plazma membrana, bez promene u citoplazmatskoj frakciji. Supstitucija insulinom povećala je nivo proteina oba transportera za glukozu u citoplazmatskoj frakciji, ali nije uticala na smanjen nivo u frakciji plazma membrana. Analiza transportera za masne kiseline je pokazala da je ekspresija gena za CD36 bila povećana u srcu pacova sa dijabetesom kao i nivo proteina u frakciji plazma membrana, što je praćeno smanjenjem nivoa ovog transportera u citoplazmatskoj frakciji. Tretman hormonom je vratio nivo iRNK za CD36 na nivo kontrole, kao i nivo proteina u citoplazmatskoj frakciji, ali nije uticao na povećan nivo u frakciji plazma membrana. Sumirano, rezultati ukazuju da se u stanju dijabetesa srce skoro u potpunosti oslonilo na korišćenje masnih kiselina, dok je preuzimanje glukoze smanjeno. Tretman insulinom je uspeo da normalizuje ekspresiju, ali ne i nivo transportera na plazma membrani. Na osnovu rezultata prikazanih u okviru ove doktorske disertacije, moţe se zaključiti da u stanju akutnog dijabetesa tipa 1 dolazi do promena u signalnom putu insulina na nivou ekspresije i/ili aktivnosti kinaza PKB/Akt i ERK1/2, što se odrazilo i na nishodne efektorne molekule u srcu. Iako supstitucija insulinom ispravlja većinu promena u aktivaciji i/ili ekspresiji ispitivanih molekula u srcu nastalih nakon izazivanja dijabetesa (Akt, NOS, GLUT4, GLUT1 i CD36), uočene promene u nivou transportnih proteina za glukozu i masne kiseline u plazma membrani nisu u potpunosti ispravljene. S obzirom da je insulinska terapija primarna terapeutska strategija u lečenju dijabetesa tipa 1, naši rezultati ukazuju da insulin sam po sebi nije dovoljan da ispravi sve metaboličke promene koje su nastale u srcu. Stoga membranski proteini koji učestvuju u transportu glukoze i masnih kiselina mogu predstavljati potencijalna mesta za primenu dodatnih terapeutika u lečenju dijabetesa, u pravcu uspostavljanja metaboličke homeostaze u srcu.

nsulin deficiency and hyperglycemia, which are the main features of Type 1 diabetes are associated with a number of endocrine and metabolic disorders and one of the most common complications is the increased incidence of cardiovascular diseases. Therefore, appropriate insulin replacement therapy is necessary in order to control the disease, as well as for the prevention of potential complications. We hypothesized that insulin deficiency and hyperglycemia may lead to changes in cardiac insulin signalling pathway which consequently may result in changes in nitric oxide production and utilization of energy substrates, and that insulin replacement is required to correct the changes. To induce experimental Type 1 diabetes, male Wistar rats were injected intraperitoneally with streptozotocin (65 mg/kg). Acute diabetes was reported to occur between 8 days and 3 weeks, and chronic diabetes within 3 weeks after streptozotocin administration. Diabetic animals were either maintained untreated, or treated with insulin (3 IU) daily for a week. For analysis, intracellular signaling molecules Akt and ERK1/2 were selected, which are important intermediary of almost all effects of insulin in the heart. Furthermore we analyzed effector molecules which are regulated through activation of Akt and ERK signaling pathways, such as eNOS and iNOS, involved in the synthesis of nitric oxide, as well as glucose transporter GLUT1 and GLUT4 and transporter of fatty acids CD36. In the present study we examined the expression and/or phosphorylation of the molecules, as well as the subcellular localization of the transporters for glucose and fatty acid in the heart. It is known that the competition between arginase and NOS for a common substrate L-arginine is a limiting factor for the production of nitric oxide. In acute phase of streptozotocin induced diabetes we observed decreased L-Arg concentration at the time when cardiac arginase activity was markedly augmented. Insulin treatment reduced the elevated arginase activity, as well as increased L-arginin concentration, indicating an important regulatory effect of insulin on arginase enzymatic activity. Streptozotocin tratment decreased cardiac eNOS phosphorylation at Ser1177 and incrased iNOS protein and mRNA expression, that were restored to control level after subchronic insulin replacement. It follows that in acute phase of Type 1 diabetes there is a shift towards metabolism of L-arginin and insulin treatment restored NOS activity. Akt mRNA expression and phosphorylation at Ser473 as well as ERK1/2 protein phosphorylation were decreased in diabetic rats. Insulin replacement restored Akt mRNA expression and phosphorylation to control level but had no effects on ERK1/2 protein phosphorylation. It can be concluded that in diabetic heart both signalling pathways are changed, but insulin treatment successful restore only PI3K/Akt. Insulin is involved in the regulation of carbohydrate and lipid metabolism in the heart. In diabetes, the metabolic changes occur in the use of energy substrates, which results in a reduced glucose uptake and increased fatty acids utilization. Diabetes has effects on redistribution of transporters from intracellular vesicles to the plasma membrane. GLUT4 and GLUT1protein/mRNA expression were decreased in diabetic rat hearts and insulin treatment restored GLUT1 expression and GLUT4 protein level. Streptozotocin treatment decreased GLUT4 and GLUT1 protein level on plasma membranes without changes in cytosol fraction. Insulin replacement augmented glucose transporters in citoplasma but the protein level on plasma membrane remained decreased. The mRNA expression of fatty acid transporter CD36 was increased as well as protein level on plasma membrane, which is accompanied by the protein reduction in cytosol. Insulin tratment restored CD36 mRNA expression and protein level in cytosol but the CD36 protein level on plasma membrane remained increased. Results indicate that diabetic heart completely depends on fatty acid utilization while glucose uptake was reduced and hormone replacement was able to normalize the expression, but not the level of the transporters on plasma membrane. In conclusion, these results indicate that PKB/Akt and ERK1/2 expression and/or activity as well as downstream target molecules are altered during streptozotocin-induced diabetes in the heart. However, although insulin repalcement corrected most of the alterations induced by diabetes in the rat heart (Akt, NOS, GLU4, GLUT1 and CD36), the levels of glucose and fatty acid transporters on plasma membrane were incompletely restored. Since, insulin therapy is the primary terapeutic strategy in the treatment of Type 1 diabetes, the results also suggest that insulin alone is not sufficient to correct all metabolic changes in the heart. Thus, membrane proteins participating in the transport of glucose and fatty acids may represent a potential targets for additional therapeutic in the treatment of diabetes, to restore cardiac metabolic homeostasis.

Prostorna distribucija i ekološke varijacije staništa hipogeičnih makromiceta (Mycota) u Srbiji

Spatial distribution and habitat variability of hypogaeous macromycetes (Mycota) in Serbia

Jedan broj gljiva iz različitih taksonomskih grupa procesom konvergentne evolucije razvio je zajedničku ekološku strategiju formiranja makroskopskih podzemnih sporokarpa. Zbog te osobine nazvane su hipogeične ili podzemne gljive. One imaju značajnu ulogu u šumskim ekosistemima kao fungalni partneri u simbiotskom odnosu sa drvećem, a ovaj odnos je obostrano od esencijalne koristi za biljku i gljivu. Takođe su i važan izvor hrane za šumske životinje. U ovoj grupi najpoznatiji su tartufi, cenjene i skupe gljive koje se koriste u ljudskoj ishrani i imaju ekonomski značaj, ali pored njih i brojne druge vrste imaju veliku i važnu ulogu u prirodi. Iako najstariji podaci o hipogeičnim gljivama na području Srbije potiču još iz 18. veka, do danas je vrlo malo poznato o njima i postoji samo petnaestak objavljenih naučnih radova i nekoliko kongresnih saopštenja, uglavnom s kraja 20. i početka 21. veka. U njima su prvenstveno obrađivani tartufi, dok o drugim grupama gotovo da nema nikakvih podataka i one su predstavljale veliku nepoznatu komponentu biodiverziteta Srbije. Jedan od razloga za mali stepen istraženosti hipogeičnih gljiva, ne samo u Srbiji već i globalno, je problematika njihovog proučavanja i pronalaženja u prirodi zbog kriptičnog, podzemnog razvoja sporokarpa na osnovu kojih je prvenstveno moguće detektovati i locirati individue gljiva. Počev od 1991. godine započeta su istraživanja hipogeičnih gljiva uz pomoć obučenih pasa, inicirana od strane dr Miroljuba Milenkovića u kojima je učestvovalo više saradnika. Na taj način formirana je zbirka koja je 2012. godine smeštena u fungarijum Prirodnjačkog muzeja u Beogradu, i od tada je nastavljeno sa njenim prinavljanjem. Najveći broj nalaza i taksona koji su u njoj deponovani nisu do sada objavljeni. Glavni cilj ove disertacije je da se rasvetli i analizira diverzitet i položaj hipogeičnih gljiva u ekosistemima Srbije, da se napravi sveobuhvatan uvid i analiza podataka, objedine znanja i na taj način doprinese i unapredi fundamentalno poznavanje i razumevanje distribucije i ekologije hipogeičnih gljiva u Srbiji i na evropskom kontinentu...

Hypogaeous fungi have evolved a common ecological strategy of forming macroscopic underground sporocarps as a result of convergent evolution. This polyphyletic group includes a number of species from different taxonomic units. They play an important role in forest ecosystems as fungal partners in essential mutualistic symbiotic associations with trees where both organisms derive benefit. Furthermore, they are an important source of food for forest animals. The truffles, the prized and expensive fungi of culinary and economic importance, are the best known of this group, however, there are other numerous species that serve a large and significant role in nature. Although the oldest data on hypogaeous fungi from the territory of Serbia date back to the 18th century, very little is known about them to this day and there are only about a dozen published original scientific articles and several conference papers, mostly dating from the late 20th and the early 21st centuries. These articles and papers primarily addressed the truffles, and offered hardly any data about other groups, which presented a large unknown component of Serbia’s biodiversity. A reason that hypogaeous fungi had not been researched to a greater degree, either in Serbia or worldwide, lies in the difficulties associated with studying and detecting these fungi in nature, due to the cryptic, underground development of sporocarps, whose presence basically allows for the detection and location of the fungal individuals. The year 1991 saw a commencement of investigation of hypogaeous fungi with the help of trained dogs, initiated by Dr Miroljub Milenković involving a number of associates. Their efforts led to the formation of a collection which was housed in the Fungarium of the Natural History Museum in Belgrade in 2012. The collection has grown since then. Most findings and taxa deposited in the Fungarium had not been published until now. The main goal of this doctoral dissertation was to research, analyse and shed light on the diversity and status of hypogaeous fungi in Serbia’s ecosystems, provide a comprehensive survey and analysis of data, consolidate data from multiple sources and contribute to and advance the fundamental knowledge and understanding of distribution and ecology of hypogaeous fungi in Serbia and Europe...

Mehanizmi uključeni u regulaciju smanjene endokrine funkcije Lajdigovih ćelija tokom starenja

The mechanisms involved in the regulation of impaired Leydig cells endocrine function during aging

Reproduktivno starenje muškaraca razvija se uporedo sa starenjem organizma, a manifestuje se značajnim promena u funkciji endokrinog sistema i smanjenim fekunditetom. U kontekstu izmenjene funkcionalnosti endokrinog sistema ističe se smanjen nivo testosterona koji zajedno sa brojnim psiho-fizičkim promenama smanjuje kvalitet života jedinke. Konstantno produženje životnog veka i potencijalno neželjena dejstva zamenske terapije androgenima usmeravaju buduća istraživanja ka boljem razumevanju molekularnih osnova muškog hipogonadizma i pospešivanju endogene produkcije androgena u starijem životnom dobu muškaraca. U skladu sa iznetim činjenicama i usmerenjma postavljeni su sledeći ciljevi istraživanja: 1. Karakterizacija staračkog hipogonadizma kod pacova soja Wistar kao modela za istraživanje reproduktivnog starenja; 2. Ispitivanje udela promena cAMP- i cGMP-signalizacije u formiranju starog fenotipa Lć. Najvažnijim rezultatima ustanovljeno je da starenje Wistar pacova prati pojava primarnog hipogonadizma koji je iskazan smanjenom produkcijom testosterona i smanjenim eksprimiranjem steroidogenih gena i proteina od dvanaestog meseca. Zajedno sa smanjenim androgenim kapacitetom poremećena je i cAMP, MAPK i NO-cGMP signalizacija kao i energetska homeostaza u Lć starog fenotipa. Tokom istraživanja ustanovljeno je da smanjen nivo cAMP nije jedini uzročnik smanjenog androgenog kapaciteta, a akutna i hronična inhibicija PDE5 povećava steroidogeni kapacitet i normalizuje energetsku homeostazu u Lć starog fenotipa. Takođe je ustanovljeno da cAMP i cGMP različito regulišu energetsku homeostazu Lć, pri čemu cGMP normalizuje parametre funkcionalnosti mitohondrija i eksprimiranje regulatora energetske homeostaze.

Reproductive aging of males develops along with the aging of the organism, and it is manifested by a significant change in the endocrine system and decreased fecundity. The decreased level of testosterone stands out in the context of the changed functionality of the endocrine system which together with numerous psycho-physical changes, reduces the quality of life of the individual. In respect with extended human life and undesirable effects of androgens replacement therapy, further investigations has been dedicated to better understanding of male hypogonadism by the molecular approach as well as endogen testosterone production during the aging. According to the presented facts we defined the two aims of the study: 1. Characterization of the age-related hypogonadism in the male Wistar rats as a model for reproductive aging, and 2. Investigation of the impact of a cAMP and a cGMP disturbances in development of aged Leydig cells. Our results showed appearance of primary hypogonadism during the aging of male Wistar rats, including impair testosterone production and impaired expression of steroidogenic genes and proteins from the 12th month of age. Beside that cAMP, MAPK and NO-cGMP signaling has been disturbed in aged Lc as well as energy homeostasis. We also showed that less production of cAMP is not the only cause of aged Lc sub-functionality and that acute and chronic inhibition of the PDE5 showed positive effect at steroidogenesis and energy homeostasis in aged Lc. Further, cAMP and cGMP differently regulated energy homeostasis of aged LC, with positive impact of the cGMP treatment on mitochondrial functionality and energy homeostasis regulators.

Beograd kao evropski grad u devetnaestom veku: transformacija urbanog pejzaža

Belgrade as a european city in the nineteenth century:urban landscape transformation

Teorijska proučavanja pejzaža kao inherentne odlike ljudskog društva, u zapadnoj civilizaciji javljaju se početkom Renesanse...

Theoretical considerations of landscapes as intrinsically characteristic of human society first appear in Western civilisation in the early Renaissance...

Duhovnost kao naracija : (o poetici proze Miodraga Pavlovića)

Spirituality as narration : (on the poetics of the prose of Miodrag Pavlovic)

Cilj ove doktorske disertacija je da prikaže impozantno stvaralaštvo živog i aktuelnog pisca srpske moderne književnosti – Miodraga Pavlovića. Osim što je postavio temelje modernog stvaralaštva, Miodrag Pavlović još uvek unosi modernu kritičku svest i traje na književnoj sceni punii 60 godina. Njegov rad obuhvata više od četrdeset knjiga poezije, dvadeset knjiga eseja i prozu obeleženu romanesknom tetralogijom. Veliko književno, teološko, antropološko i filozofsko znanje ovog stvaraoca doprinosi kvalitetu i književnoj kulturi ne samo našeg, već i svetskog podneblja. Tehnički, rad se sastoji iz dva dela. Prvi deo obuhvata pojedinačna dela ( pregled romana), a drugi deo je pogled na opšta pitanja u vezi sa apokalipsom, antropologijom, simbolikom, tj. obuhvata opšta pitanja poetike i duhovnosti kao naracije u stvaralaštvu Miodraga Pavlovića. U osnovi celokupnog stvaralaštva stoji mit i obred uz antropološki pogled na religiju, ali i ezoterna izučavanja, filozofija i tradicija. Pitanje Apokalipse okosnica je svih tih razmišljanja i učenja. Udaljenost od dogmatskih razrešenja vodila je kompleksnijim razmišljanjima, različitim vizijama i oblikovala nova umetnička dela koja su potvrđivala činjenicu da je u svetu sve povezano. Jedinstven u idejama uspeo je da rekonstruiše daleke obredne radnje, istorijske događaje i samim tim ih približio savremenom čitaocu. Uvodi drugačije značenje novog i modernog ukazujući da je svet samo jednom bio nov, a sve što se kasnije odvijalo samo je interpretiranje starog.

The aim of the doctoral dissertation is to present the work of a great living writer of Serbian modern literature - Miodrag Pavlovic. Having laid the foundations of modern literature writing, Miodrag Pavlovic has been imbued with modern critical consciousness for the last sixty years, engendering some of the most prominent literary achievements in Serbia. The literary work consists of more than 40 books of poetry, 20 books of essays as well as prose marked by romanesque tetralogy. The comprehensive knowledge of Miodrag Pavlovic and his ponderings upon literature, anthropology and philosophy make contributions to both the quality and literary culture of both our and the world’s heritage. Technically speaking, there are two parts of the thesis. The first part implies the individual works (the novels), and the second part reflects the views on apocalypse, anthropology, symbolics related to the general issues of poetics and spirituality as narration in Miodrag Pavlovic’s work. The myth and the rite make the essence of his work, coupled with an anthropological view of religion, esoteric studies, philosophy, and tradition. The apocalyptic issue presents the framework for all the ponderings and studies. The distance from dogmatic solutions led to even more complex ponderings, various visions and thus shaped new artistic works which confirmed the universal connection between all things in the world. Owing to his ideas, Miodrag Pavlovic has succeeded in reconstructing the ancient rituals, historical events bringing them closer to a contemporary reader. Pavlovic introduces a fairly different meaning of the new and the modern implying that the world was new only once, everything that was to occur afterwards has been a mere interpretation of the same.

Analiza regulacije proteinaznih gena prirodnog izolata: Lactococcus lactis subsp. lactis BGIS29

Analyses of prt genes regulation in the natural isolate Lactococcus lactis subsp. lactis BGIS29

Prirodni izolat Lactococcus lactis subsp. lactis BGIS29 produkuje kiselu proteinazu PI tipa i bakteriocin IS29. Prisustvo kazitona u minimalnom medijumu (MM) ima specifičan uticaj na regulaciju aktivnosti gena za proteinazu soja BGIS29. Transkripcione genske fuzijc sa E. coli ß-glukuronidaznim genom (gusA) su korišćene za izučavanje medijum zavisne ekspresije promotora prt gena (VpnP i PprtM). Merenjem aktivnosti enzima ß-glukuronidaze i kvantitativnom Northern blot analizom je pokazano da je aktivnost oba (VprtP i ?pnM) promotora kontrolisana kazitonom na transkripcionom nivou, ali je stepen njihove regulacije različit. Kada PpnF promotor kontroliše ekspresiju gusA gena, aktivnost enzima ß-glukuronidaze lineamo opada sa porastom koncentracije kazitona u medijumu za rast bakterija (0.1% do 2%). U slučaju PpnM promotora regulacija nije tako jaka kao u slučaju PpnP promotora. Visoka koncentracija kazitona prisutna u MM ima inhibitoran efekat na aktivnost P prtM promotora, ali nema lineame korelacije. Deleciona analiza regulatomog regiona prt gena je pokazala da je celokupna informacija neophodna za punu aktivnost i regulaciju P pnM promotora sadržana u sekvenci regiona dužine 180 bp koji sadrži oba prt promotora. Izucavan je uticaj nukleotidne sekvence na krivljenje DNK. Eksperimenti pokretljivosti DNK fragmenata u gelu su korišćeni u kombinaciji sa kompjuterskom predikcijom potencijalnih zakrivljenih regiona DNK. Kompjuterska predikcija krivljenja DNK je pokazala da region uzvodno od -35 i -10 regiona Pprtp promotora sadrži potencijalni centar krivljenja. Eksperimenti pokretljivosti DNK fragmenata u gelu su pokazali da je PCR fragment veličine 350 bp, koji sadrži kompletan regulatomi region, zakrivljen. Koeficijent retardacije (R) je 1.2 u odsustvu MgCl2 i 1.4 u prisustvu MgCl2. Kraći PCR fragment (270 bp) koji sadrži oba prt promotora je, takode, zakrivljen, R je 1.1 u odsustvu MgCl2 i 1.5 u prisustvu MgCl2. Inaktivacija (“knock-out”) potencijalnog regulatomog gena, prtR, je uradena insercijom plazmida pGGiostOzISN/ u hromozom soja Lactococcus lactis NZ9000. Merenjem aktivnosti enzima ß-glukuronidaze je pokazano da nema aktivnosti PpnP promotora u ”knock-out” soju. Rezultati ukazuju da je potencijalni regulatomi gen aktivator.

Natural isolate Lactococcus lactis subsp. lactis BGIS29 produces the Pi-type proteinase and bacteriocin IS29. The presence~of uasitone in chemically defined medium (CDM) has a specific influence on the regulation of the proteinase activity in the strain BGIS29. Transcriptional gene fusion’s with E. coli ß-glucuronidase gene (gusA) were used to study medium dependent expression of both ¥pnP and ¥prtM promoters. The ß-glucuronidase.assays and the quantitative Northern blot analysis showed that activities of PpnP and VpnM promoters are controlled by casitone at the transcriptional level but their regulation is occurring in different manner. When the ¥prtP promoter controlled the gusA gene expression, the ß-glucuronidase activity was gradually decreasing with increase of casitone in the growth medium (0.1% to 2%). In the case of the ¥ pnM promoter the regulation is not that strong as in the case of the ¥ prtP promoter. Higher concentrations of casitone present in CDM had an inhibitory effect on the ¥pnM promoter but there is no linear correlation. Deletion analysis of regulatory region of the prt genes revealed that all information needed for full activity and regulation of the ¥ prtM promoter is retained within 180-bp region that contains both the PortP and P^^promoters sequences. The influence of nucleotide sequence context on the DNA bending was investigated. Gel mobility experiments are used together with the computer-assisted prediction of a putative DNA bending. Computer-assisted prediction of the DNA bending showed that the region upstream of the —35 and -10 regions of the ¥prtP promoter contains a putative center of the bending. Gel mobility experiments revealed that the PCR fragment of 350 bp containing intact regulatory region is curved, the retardation value (R) was 1.2 in the absence of the MgCl2 and 1.4 in the presence of the MgCl2. Shorter PCR fragment (270 bp) containing both prt promoters is also curved, R were 1.1 and 1.5 in the absence and presence of MgCl2, respectively. A knockout of a putative regulatory gene, prtR, was done by insertion of the plasmid pG+host9::IS.S7 into chromosome of the strain Lactococcus lactis NZ9000. There was no the ß-glucuronidase activity controlled by ¥prtP promoter in the knock-out strain. The results indicates that a putative regulatory gene could be an activator.

Uticaj atrazina i bisfenola A na funkciju granuloza ćelija ovarijuma pacova

Effects of atrazine and biphenol A on the rat ovarian granulosa cells

Atrazin (ATR), jedan od najčešće korišćenih herbicida, i bisfenol A (BPA), hemikalija koja ulazi u sastav plastike, pripadaju grupi endokrinih ometača koji, između ostalog, ispoljavaju negativne efekte na ženski reproduktivni sistem, prvenstveno na nivou steroidogeneze ovarijumskih granuloza ćelija. Cilj ove doktorske disertacije je da proširi saznanja o uticaju ATR i BPA na steroidogenezu granuloza ćelija, ukaže na nova mesta delovanja ovih endokrinih ometača, kao i da definiše mehanizme preko kojih ATR i BPA utiču na steroidogeni i ovulacioni potencijal granuloza ćelija. In vitro tretman primarne kulture granuloza ćelija sa 20 µM ATR doveo je do smanjene produkcije estradiola i povećane produkcije progesterona, kao i promena na nivou ekspresije gena za protein uključene u steroidogenezu i diferencijaciju granuloza ćelija (aromataza, LHR, StAR, dezmolaza) i ovulaciju (AREG, EREG, PGR). Mehanizam delovanja ATR uključuje promene u aktivnosti cAMP/PKA, ERK1/2/CEBPβ i Akt signalnih puteva. In vivo tretman sa 50 mg/kg telesne mase ATR doveo je do smanjene ekspresije gena uključenih u kontrolu ovulacije (EREG, PGR) i izostanka ovulacije kod nezrelih ženki pacova tretiranih gonadotropinima. In vitro tretman nezrelih granuloza ćelija sa 100 µM BPA doveo je do smanjenog stvaranja progesterona putem narušavanja homeostaze holesterola, što se ogleda u sekvestraciji holesterola u perinukleusnom prostoru, smanjenoj ekspresiji gena za protein ABCA1, zaduženog za efluks holesterola, i povećanoj ekspresiji StAR proteina uključenog u transport holesterola i SREBP-1 proteina neophodnog za sintezu holesterola. Dobijeni rezultati ukazuju na nove mehanizme preko kojih ATR i BPA utiču na funkciju granuloza ćelija ovarijuma, kao i na potencijal za razvoj bolesti ovarijuma.

Atrazine (ATR), a widely used herbicide, and bisphenol A (BPA), a chemical used in manufacturing of the polycarbonate plastics, belong to the group of endocrine disrupting chemicals. ATR and BPA exert adverse effects on ovarian granulosa cells’ steroidogenesis. The aim of this study was to expand the knowledge on the effects of ATR and BPA, reveal new sites of ATR and BPA action in the ovary, and define the mechanisms through which ATR and BPA affect steroidogenic and ovulatory potential of granulosa cells. In vitro treatment of the primary culture of granulosa cells with 20 µM ATR caused a decrease in estradiol production, an increase in progesterone production, as well as changes in the expression level of the genes involved in steroidogenesis and differentiation of granulosa cells (Cyp19a1, Lhr, Star, Cyp11a1) and ovulation (Areg, Ereg, Pgr). The mechanism of ATR action involved changes in the activity of cAMP/PKA, ERK1/2/CEBPβ and Akt signaling pathways. In vivo treatment with 50 mg/kg body weight ATR reduced the expression level of the ovulatory genes (Ereg, Pgr) and blocked ovulation in the gonadotropin-treated immature female rats. In vitro treatment of immature granulosa cells with 100 µM BPA decreased progesterone production and altered cholesterol homeostasis, as evident by cholesterol sequestration to the perinuclear area, decrease in the expression level of the gene involved in cholesterol efflux (Abca1), and increase in the level of proteins involved in cholesterol transport (StAR) and biosynthesis (SREBP-1). Results reveal the novel mechanisms through which ATR and BPA impair function of ovarian granulosa cells, as well as the potential relationship between exposure to these endocrine disruptors and development of ovarian diseases.

Promjenljivost srčanog ritma i oštećenje DNK u hemocitama kod mediteranske dagnje Mytilus galloprovincialis L. kao biomarkeri zagađenja mora

Heart rate variation and DNA damage in haemocytes of the mediterranean mussel Mytilus galloprovincialis L. as biomarkers of marine pollution

Integrisani pristup biomonitoringa koji uključuje bateriju biotestova, omogućava pouzdaniju detekciju različith vrsta organskog i neorganskog zagađenja u cilju procjene stanja u morskom ekosistemu. U ovom radu je praćen uticaj hemijskih stresora na srčani ritam mediteranske dagnje pri kratkotrajnim tretmanima ex situ. Pošto biomarkeri na ćelijskom i molekularnom nivou imaju veću specifičnost, u cilju unaprijeđenja pouzdanosti i praktične primjenljivosti srčanog ritma kao fiziološkog biomarkera, uradjeni su i produženi tretmani dagnji, takođe ex situ, u kojima su uporedno analizirani srčani ritam i oštećenja DNK u hemocitama mediteranske dagnje. Pri kratkotrajnim tretmanima, ispitivan je uticaj promjene u salinitetu morske vode na srčani ritam dagnji koji je praćen fotopletizmografski, fiber-optic metodom. Takođe, ispitivan je odgovor srčanog ritma dagnje pri izlaganju kadmijum hloridu, dizel gorivu, naftnom disperzantu Superdispersant-25 i dispergovanom dizelu, pri čemu je potvrđena sposobnost dagnji da detektuju stres u ranoj fazi. Prilikom produženih tretmana dagnji, uporedno sa praćenjem srčanog ritma, procijenjen je genotoksični potencijal kadmijum hlorida kao model mutagena, dizel goriva, Superdispersant-25, benzo(a)pirena i tributil kalaj hlorida kao zagađivača čija kontrola je od velikog značaja. Uz komet test koji je služio za procjenu oštećenja DNK, genotoksični potencijal tributil kalaj hlorida, analiziran je mikronukleus testom i prokariotskim SOS/umuC testu in vitro. Dobijeni rezultati su pokazali da je dispergovani dizel mnogo opasniji po školjke usljed veće dostupnosti njegovih sastojaka. Rezultati praćenja srčanog ritma pri tretmanima tributil kalaj hloridom mogu se povezati sa podacima dobijenim u genotoksikološkim testovima što ukazuje da bi se srčani ritam dagnji mogao koristiti kao pouzdan biomarker u detekciji zagađenja porijeklom od organokalajnih jedinjenja. Na osnovu rezultata dobijenih pri kratkotrajnim i produženim tretmanima organskim i neorganskim zagađivačima, srčani ritam mediteranske dagnje se može preporučiti za korišćenje kao sastavni dio integrisanog pristupa u biomonitoringu zagađenja mora.

reliable detection of different types of organic and inorganic pollutants which would improve the evaluation of the state of the marine ecosystem. In this study, the influence of short-term treatments by chemical stressors on the heart rate of the Mediterranean mussel was monitored ex situ. Since cellular and molecular biomarkers are characterized by higher specificity, extended treatments for comparative analyses of the heart rate and DNA damage in haemocytes were performed to improve reliability and applicability of the Mediterranean mussel’s heart rate as a physiological biomarker. Fiber-optic method, based on photoplethysmography, was applied for studying the influence of variations in seawater salinity on the heart rate of mussels. The influence of cadmium chloride, diesel oil, oil dispersant Superdispersant-25 and dispersed diesel oil was studied as well, whereby the ability of mussel’s heart rate to detect stress at an early stage was confirmed. Comparative analyses of mussel’s heart rate and genotoxic potential of cadmium chloride as a model mutagen, diesel oil, Superdispersant-25, benzo(a)pyrene and tributyltin as pollutants of importance to control, in extended treatments were assessed. Comet assay was used for the assessment of DNA damage. Moreover, genotoxic potential of tributyltin was assessed in vitro by micronucleus test and prokaryotic SOS/umuC assay. The results indicated that dispersed diesel oil was more harmful for mussels due to enhanced availability of its compounds. Heart rate results obtained in tributyltin chloride treatments can be linked to the data obtained in genotoxicological assays indicating that musels‘s heart rate could be used as a reliable biomarker for detection of organotin pollution. The results obtained in short-term and extended treatments by organic and inorganic pollutants indicated that heart rate of the Mediterranean mussel can be included as a part of the integrated approach of marine pollution biomonitoring.

Sezonske promene u sadržaju toksičnih metala u zemljištu i listovima zimzelenih vrsta Prunus laurocerasus L., Buxus sempervirens L. i Mahonia aquifolium (Purch) Nutt. na području grada Beograda

Seasonal changes in the content of toxic metals in the soil and the leaves of evergreen species Prunus laurocerasus L. , Bruxus sempervirens L. and Mahonia aquifolium (Purch) Nutt. in the city of Belgrade

Urbana sredina se od okolnih ruralnih zona razlikuje u pogledu klimatskih uslova, kao i po kvalitetu vazduha. Pored toga, zemljišta podležu uticaju antropogenog faktora, pa se u zemljištima urbano-industrijskih zona u različitim koncentracijama akumuliraju toksični teški metali, nitrati, pesticidi i organske zagađujuće supstance. Različite vrste biljaka ispoljavaju različit stepen osetljivosti na prisustvo određenih vrsta zagađujućih supstanci, kao i na njihovu povećanu koncentraciju. Iz tog razloga sprovodi se biološki monitoring koji obuhvata određivanje reakcija biljaka na promene u koncentraciji zagađujućih materija u spoljašnjoj sredini koje je zasnovano na praćenju vidljivih promena na biljkama kao i detekciji i utvrđivanju njihove količine u biljnim organima analitičkim metodama u laboratorijskim uslovima. Zimzelene, širokolisne vrste biljaka, Prunus laurocerasus L., Buxus sempervirens L. i Mahonia aquifolium (Pursh) Nutt. se već duži niz godina uspešno gaje u urbanim uslovima. I pored toga što veliki broj jedinki ovih vrsta raste u neposrednoj blizini visokofrekventnih saobraćajnica i izvora aerozagađenja, višegodišnji listovi ovih biljaka su bez vidljivih znakova oštećenja. Na osnovu tih činjenica definisani su i ciljevi ove disertacije: 1) Utvrđivanje osnovnih karakteristika staništa ispitivanih vrsta biljaka na području grada Beograda, uključujući podatke o tipu urbanog biotopa kojeg naseljavaju, pedološkoj podlozi i klimi; 2) Određivanje osnovnih hemijskih osobina (pHH2O, pHKCl, procenat organske materije) zemljišta na urbanim lokalitetima na području grada Beograda, kao i prirodnim staništima ispitivanih vrsta biljaka; 3) Određivanje koncentracije Mg, Ca, Na, Fe, Mn, Ni, Zn, Cr, Co, Cu, Cd, Pb, V u zemljištu na urbanim lokalitetima na području grada Beograda i prirodnim staništima ispitivanih vrsta biljaka; 4) Ispitivanje koncentracije navedenih makro elemenata i toksičnih (teških) metala u listovima biljaka P. laurocerasus L., B. sempervirens L. i M. aquifolium (Pursh) Nutt. sa urbanih lokaliteta u Beogradu i sa prirodnih staništa; 5) Određivanje bioakumulacionog faktora (BF) u cilju određivanja efikasnosti biljaka da usvajaju i akumuliraju hemijske elemente u listovima; 6) Procena kapaciteta ispitivanih zimzelenih biljnih vrsta za toleranciju ili bioakumulaciju određenih toksičnih metala u cilju utvrđivanja i otkrivanja novih indikatorskih, tolerantnih i/ili potencijalnih akumulatorskih zimzelenih biljnih vrsta. Za sakupljanje uzoraka biljaka i zemljišta, odabrano je šest širih lokaliteta (uključujući 18 užih lokaliteta) u užoj gradskoj zoni grada Beograda, kao i tri lokaliteta sa prirodnih staništa ispitivanih vrsta biljaka. Uzorci listova ispitivanih vrsta biljaka i zemljišta sakupljani su sa svakog lokaliteta četiri puta tokom 2012. godine, odnosno po jednom u svakoj sezoni. Ispitivanja biljnog materijala i zemljišta su obuhvatila sledeće: 1) određivanje pH zemljišnog rastvora; 2) određivanje količine organskog ugljenika i humusa u zemljištu; 3) određivanje ukupnog sadržaja ispitivanih elemenata u zemljištu i biljnom materijalu. Određivanje stvarne i izmenljive pH vrednosti zemljišnog rastvora, količine organske materije, izmenljivog Ca, Mg, Na, kao i svih ispitivanih makro i toksičnih elemenata u uzorcima zemljišta i biljaka urađene su prema standardnim procedurama. Analize svih uzoraka zemljišta i biljaka su urađene u tri ponavljanja, a dobijeni podaci su statistički obrađeni. Numeričke deskriptivne mere uključile su srednju vrednost (M) i standardnu devijaciju (SD). Rezultati su prikazani tabelarno i grafički. Za analizu međusobnih odnosa elemenata u zemljištu i biljkama korišten je Spirman-ov koeficijent korelacije (rs). Da bi se utvrdio odnos između teških metala u zemljištu, kao i njihovi mogući izvori, korišćen je Pirson-ov koeficijent korelacije i analiza glavnih komponenti (PCA - Principal Component Analysis). Na osnovu podataka dobijenih hemijskim analizama 72 uzorka zemljišta sakupljena tokom četiri uzastopne sezone 2012. godine u užoj gradskoj zoni Beograda kao i na prirodnim, nezagađenim lokalitetima ispitivanih vrsta biljaka, utvrđeno je da se izmerena pH vrednost zemljišta u gradu kreće od blago kisele do blago bazne reakcije (pHH2O 6,11 − 8,16; pHKCl 5,78 − 7,48), a sadržaj organskih jedinjenja oko 5% što se smatra umerenim i karakterističnim za zemljišta koja se odlikuju relativno brzom razgradnjom organskih ostataka. Utvrđeno je da je značajno viši sadržaj Mg u zemljištu bio prisutan u proleće i leto, a Ca tokom zimskog perioda. Prosečne koncentracije mangana, nikla, cinka, kobalta, kadmijuma i olova u ispitivanim uzorcima zemljišta iz Beograda su bile u nivou vrednosti koje su konstatovane za druge gradove u Evropi i svetu, izuzev vanadijuma, čija je koncentracija u ispitivanim uzorcima zemljišta ispod prosečnih vrednosti u svetu. U poređenju sa drugim gradovima u Srbiji (Niš, Smederevo, Požarevac, Subotica i Novi Sad) koncentracije većine ispitivanih elemenata u zemljištu Beograda su nešto više. Sadržaj toksičnih ili teških metala i vanadijuma u uzorcima zemljišta je u različitoj meri varirao u zavisnosti od lokaliteta i sezone. Na osnovu statističkih analiza dobijenih podataka utvrđeno je da se u površinskom sloju zemljišta u Beogradu, s jedne strane nalazi veća koncentracija toksičnih ili teških metala koji se u literaturi označavaju kao „urbani metali“ (Pb, Zn, Cu, Cd), a sa druge strane se nalaze metali koji vode poreklo od matične stene (Ni, Cr, Mn). Analizom biljnog materijala ispitivanih vrsta B. sempervirens, M. aquifolium i P. laurocerasus utvrđeno je da se koncentracija pojedinih elemenata razlikovala u odnosu na biljnu vrstu, kao i u odnosu na različite urbane i prirodne lokalitete. Najveće koncentracije Mn, Ni, Zn, Cr i Cu, zabeležene su u listovima vrste M. aquifolium ali je uočeno i najveće variranje ovih elemenata tokom sezona. Koncentracije Co, Cd, Pb i V su bile prilično ujednačene u listovima kod sve tri ispitivane vrste. Utvrđeno je, takođe, da su koncentracije Fe i Mn uglavnom bile u granicama normalnih vrednosti prikazanih u literaturi, pri čemu su veće koncentracije u listovima bile izmerene u jesen i zimu. U listovima ispitivanih biljnih vrsta zabeležene su visoke koncentracije Ni tokom proleća, a posebno visoke u uzorcima sakupljenim u letnjem periodu, a koje su i po nekoliko puta veće od normalnih vrednosti. Koncentracije Pb, Cd i Cr u listovima sve tri ispitivane vrste biljaka bile su iznad vrednosti koje se smatraju normalnim, a najveće koncentracije u listovima su bile tokom proleća i leta. U listovima sve tri ispitivane vrste Cu ima najviše tokom jeseni i zime i to iznad normalnih vrednosti za biljke, a u nekim slučajevima su čak na granici toksičnih vrednosti. Koncentracija Zn u listovima ispitivanih vrsta biljaka se kretala u okviru normalnih vrednosti prikazanih u literaturi i za ovaj elemenat nije ustanovljen jasan trend promene tokom sezona. Kobalt nije identifikovan u svim uzorcima listova ispitivanih vrsta biljaka, ali u uzorcima u kojima je pronađen, je iznad normalnih vrednosti i veoma blizu količinama koje se smatraju toksičnim za većinu biljaka. Sadržaj V u listovima ispitivanih vrsta biljaka identifikovan je samo u tri uzorka listova i to sva tri u proleće, u granicama normalnih vrednosti. Za ispitivanu vrstu B. sempervirens izračunate vrednosti indeksa bioakumulacije bile su veće od jedan IBA>1 za Cd, Cu, Pb i Co, na nekoliko urbanih lokaliteta. Za ispitivanu vrstu M. aquifolium IBA>1 za Cd i ispitivanu vrstu P. laurocerasus, IBA>1 za Cd i Cu, takođe na nekoliko urbanih lokaliteta. Na osnovu dobijenih rezultata vrste B. sempervirens, M. aquifolium i P. laurocerasus mogu se smatrati za neke od ispitivanih elementa indikatorima, a za većinu ekskluderima zbog čega su veoma pogodne za uspešno gajenje na metalima opterećenim zemljištima urbanih ekosistema.

Urban environment of the surrounding rural areas differ in terms of climate conditions, as well as air quality. In addition, the land is subjected to the influence of anthropogenic factors, resulting with accumulations of toxic heavy metals, nitrates, pesticides and organic pollutants in different concentrations in the soils of urban- industrial zones. Different types of plants exhibit varying degrees of sensitivity to the presence of certain types of pollutants, as well as their increased concentration. Biological monitoring is established to determine the reaction of plants to changes in the concentration of pollutants in the environment, based on monitoring of visible changes to the plant as well as the detection and determination of the amounts of the plant organs using analytical methods in laboratory conditions. Evergreen, broadleaf plant species, Prunus laurocerasus L., Buxus sempervirens L. and Mahonia aquifolium (Pursh) Nutt. have for many years successfully grown in urban conditions. Despite the fact that a large number of units of these species is growing near high-traffic routes and sources of air pollution, perennial leaves of these plants have no visible signs of damage. The objectives of this thesis are based on these facts and they include: 1) Identification of the main characteristics of the habitats of investigated species of plants in the city of Belgrade, including data on the type of urban biotope they inhabit, and pedological substrate and climate: 2) Determination of basic chemical properties (pHH2O, pHKCl, the concentration of organic matter) of land in urban localities in the city of Belgrade, as well as the natural habitats of the studied species of plants; 3) Determination of Mg, Ca, Na, Fe, Mn, Ni, Zn, Cr, Co, Cu, Cd, Pb, V in the soil in urban localities in the city of Belgrade and the natural habitats of the studied species of plants; 4) Testing concentrations of the major elements and toxic (heavy) metals in the leaves of plants P. laurocerasus L., B. sempervirens L. and M. aquifolium (Pursh) Nutt. from urban localities in Belgrade and natural habitats; 5) Determination of the Bioaccumulation factor (BF) in order to determine the efficiency of plants to acquire and accumulate chemical elements in the leaves; 6) Assessment of capacity of examined evergreen plants for tolerance or bioaccumulation of certain toxic metals in order to determine and discover new indicating, tolerant and/or potentially accumulating evergreen plant species. Six wider localities, including 18 immediate localities, in the inner city zone of Belgrade, were selected for the collecting of samples of plants and soil. Three sites natural habitats of the studied plant species were also selected. Leaf samples of the tested species of plants and soil were collected from each site four times during 2012, once in each season. Tests of plant material and soil included the following: 1) determination of the pH of the soil solution; 2) determination of the amount of organic carbon and humus in the soil; 3) determination of the total content of analyzed elements in soil and plant material. Determination of real and exchangeable pH value of the soil solution, the amount of organic matter, exchangeable Ca, Mg, Na, as well as all investigated macro and toxic elements in soil and plants were conducted according to standard procedures. Analyses of samples of soil and plants have been done in triplicate, and data were statistically analyzed. Numerical descriptive measures included the average (M) and standard deviation (SD). The results are presented in tables and graphs. Spearman's correlation coefficient (rs) was used for the analysis of mutual relations of elements in soil and plants. To determine the relationship between the heavy metals in the soil, and their possible sources, we used the Pearson's correlation coefficient and principal components analysis (PCA - Principal Component Analysis). Based on the data obtained by the chemical analysis of 72 soil samples collected in four consecutive seasons in 2012 in the inner zone of Belgrade, as well as on natural, unpolluted sites investigated species of plants, it was found that the measured pH value of land in the city ranges from slightly acidic to slightly basic reactions (pHH2O 6.11 to 8.16; pHKCl 5.78 to 7.48), and the content of organic compounds, about 5%, which is considered moderate and characteristic of land, which is characterized by a relatively rapid decomposition of organic residues. It was found that a significantly higher content of Mg was present in the soil in the spring and summer, and Ca during the winter period. Average concentrations of manganese, nickel, zinc, cobalt, cadmium and lead in soil samples from Belgrade were at the level of values that are found for other cities in Europe and the world, with the exception of vanadium, whose concentration in soil samples is below the average values in the world. Compared with other cities in Serbia (Niš, Smederevo, Požarevac, Subotica and Novi Sad), the concentration of the majority of tested elements in the soil of Belgrade are slightly higher. The content of toxic or heavy metals and vanadium in soil samples in varying degrees varied depending on location and season. The statistical analysis of the data revealed that the subsurface layers of soil in Belgrade, on one side have a higher concentration of toxic or heavy metals in the literature as "urban metals" (Pb, Zn, Cu, Cd), and on the other side are metals that originate from bedrock (Ni, Cr, Mn). The analysis of plant materials of studied species B. sempervirens, M. aquifolium and P. laurocerasus has shown that the concentration of individual elements differ in relation to plant species, as well as in relation to various urban and natural sites. The highest concentrations of Mn, Ni, Zn, Cr and Cu have been reported in leaves of M. aquifolium, however the greatest variation of these elements during the season has also been noted. The concentrations of Co, Cd, Pb, and V were quite uniform in the leaves of all three species. It was also found that the concentrations of Fe and Mn were mainly within normal limits shown in the literature, with higher concentrations in the leaves were measured in autumn and winter. The leaves of tested species have been recorded high concentrations of Ni in the spring, a particularly high in samples collected in the summer, which are several times higher than the normal value. Concentrations of Pb, Cd and Cr in the leaves all three types of plants were above levels that are considered normal, and the largest concentration in leaves were during spring and summer. Regarding copper, the leaves of all three types have most Cu in autumn and winter and above the normal range for the plants, and in some cases are even on the verge of toxic values. The concentration of Zn in the leaves of tested species of plants ranged within the normal range shown in the literature for this element and a clear trend of changes has not been established during the season. The cobalt has not been identified in most leaf samples of examined species of plants, but where found it is above normal values and very close to the amounts which are considered to be toxic to most plants. The content of the leaves of tested species of plants were identified only in three samples of leaves and all three in the spring, within the limits of normal. For the test type B. sempervirens, the calculated index values of bioaccumulation were greater than one IBA> 1 for Cd, Cu, Pb and Co, at several urban localities. For the test of the M. aquifolium IBA> 1 for Cd and tested type of P. laurocerasus, IBA> 1 for Cd and Cu, also at a few urban localities. Based on results, B. sempervirens, M. aquifolium and P. laurocerasus may be considered for some of the tested component as indicators, and for most as excluders which makes them very suitable for the successful cultivation at metal laden soils of urban ecosystems.

Prostorni kontekst sinematičke premise arhitekture

Spatial context of the cinematic aspect of architecture

Ovo istraživanje je objedinilo nekoliko sinematičkih aspekata i ponudilo jedan mogući način posmatranja fenomena urbanog prostora kroz sinematički objektiv. Rezultat je testiranja i verifikacije sinematičkog aspekta arhitekture u konekciji sa istraživanjem arhitektonskog projektovanja. Osim što obezbeđuje suštinski doprinos savremenoj teoriji arhitekture i urbanih praksi kroz postavku okvira budućih istraživanja, ova disertacija predstavlja kritiku tradicionalnih teorija u nastojanju da odgovori na urgenatna pitanja istraživača o novim načinima povezivanja teorijskih i praktičnih stanovišta. Uspostavljeni istraživački okvir, pre svega, obezbeđuje fleksibilnost u radu na operativnoj funkciji sinematičkog aspekta arhitekture u teorijskom okviru savremenog koncepta metodologije arhitektonskog projektovanja. U radu sa vizuelnim komponentama urbanog okruženja, koje kasnije prerasta u nekoliko aspekata dejstva vizuelno-imaginativnih otkrića filmskih režisera na promenu prostornih koncepcija, ovo istraživanje prevazilazi tradicionalni pogled na arhitekturu imenujući je umetnošću – umetnošću oblikovanja prostora. U ovom kontekstu i uspostavljenoj relaciji sa filmom već od prvih koraka tehnologije pokretne slike sa početka XX veka, pokrenuto je preispitivanje perceptivnih načela, iskustva arhitektonskog prostora, i otvorena relacija koncepta kretanja prema arhitektonskim i filmskim elementima i principima projektovanja. U tom okviru kroz koji arhitektura, poput svake druge umetnosti, usvaja sopstvenu vrednost i odnos prema svetu vizuelno– imaginativnom manipulacijom prostora, percepcije, programa i vizuelnosti, pokazuje na koji način dejstvo filma utiče na transformaciju arhitektonskih koncepcija prostora u sinematički događaj. Bavljenje vizuelno-imaginativnim otkrićima filmskih režisera i arhitekata i istraživanje percepcije prostora je u žiži interesovanja istraživača i čini didaktičko–edukativni motivator i inicijator preosmišljavanja IV arhitektonske prakse koji se predlaže u radu. Prostorni kontekst sinematičke premise arhitekture istražen je u uslovima u kojima aktuelne promene u savremenom arhitektonskom diskursu možemo pratiti poravnanjem odnosa arhitekture i filma sa njihovim metodološkim i didaktičkim pristupima...

This doctoral dissertation offers an observation of the urban space phenomena through the cinematic lens as a trial to testing and verification of the cinematic aspect of architecture. Beside the essential contribution to the contemporary theory of architecture, this research has attempted to discover the operational function of the cinematic aspect of architecture within the theoretical and methodological framework of the contemporary architectural design methodology. The research goes well beyond the traditional view of architecture to appoint it art - the art of shaping space. It is this framework that enables architecture to propel a review of perceptual principles, the experience of architectural space, and to open up a relationship between the concept of movement and the design elements and principles of both film and architecture. By providing an imaginative manipulation of space, perception, program configurations and visuality, architecture, like any other art, adopts its own value and relationship to the world. The dissertation offers one possible view of the consequences of such manipulations arising after the cinematic apparatus exerted an influence on the transformation of the spatial conceptions into a cinematic event. Dealing with modifications of the perception of space, visual and imaginative discoveries of filmmakers and architects is a focal research interest in terms of the didactic and educational motivator and the reassessment of architectural practice. Spatial context of the cinematic aspect of architecture was dramatically challenged by ongoing changes in contemporary architectural discourse which can be traced and investigated by harmonization of the relationship between architecture and film with their methodological and didactic approaches. This dissertation indicates various effects of the relationship between architecture and film, demonstrates points of their connection and attaches them to the current state of contemporary architectural discourse which was labeled by concept of movement, transformability, as well as to the manifestation of these phenomena. Launching of an innovative approach to the contemporary architectural design strategy was conditioned by the interpretation of these phenomena through a concept of architecture. After verifying the cinematic aspect of architecture that provided general conclusions about the relationship between the two disciplines, this research continued by investigating into a potential innovation in the theoretical and methodological framework of architectural design. The research has shown significant results in the possibility to improve and upgrade the established values of urban practices, precisely, Cullen`s urban strategy...

Definisanje prediktivnih faktora biološkog ponašanja folikulskih tumora štitaste žlezde

Definition of biological behavior predictive factors for follicular thyreoid tumors

Prema raspoloţivim dijagnostičkim metodama nije moguće preoperativno razlikovati benigne od malignih folikulskih tumora štitaste ţlezde, a najčešće ni intraoperativno zbog veoma slične ćelijske morfologije folikulskih adenoma i folikulskih karcinoma i nemogućnosti citološkog dokaza invazije kapsule ili krvnih sudova karakteristične za folikulske karcinome. U ovoj studiji, traţili smo moguće prediktivne faktore biološkog ponašanja odnosno maligniteta kod pacijenata s folikulskim karcinomom štitaste ţlezde koji bi omogućili ispravnu selekciju pacijenata za hirurško lečenje, a potom i izvoĎenje adekvatnog tipa operacije kod bolesnika s folikulskom tireoidnom neoplazmom. Metod: Ovom retrospektivnom studijom su bili obuhvaćeni svi pacijenti operisani zbog postojanja folikulskog tumora štitaste ţlezde u Centru za endokrinu hirurgiju Kliničkog Centra Srbije u Beogradu tokom petogodišnjeg perioda (2008-2012). U istraţivanje su uključena 263 operisana pacijenta. Na osnovu definitivnog histopatološkog nalaza ispitanici su podeljeni u dve grupe; folikulske adenome (97) i folikulske karcinome (166). Najvaţnije demografske i kliničke karakteristike analizirane su univarijantnom i multivarijantnom logističkom regresionom analizom. Potom je formiran nomogram tj. dvodimenzionalni dijagram koji omogućava pribliţno grafičko izračunavanje verovatnoće postojanja maligniteta kod pacijenata s dijagnozom folikulske lezije štitaste ţlezde odnosno primenu rezultata ovog rada u praksi. Rezultati: U grupi pacijenata operisanih zbog folikulskog adenoma (19 osoba muškog i 78 ţenskog pola) starosna dob je iznosila 19-79 godina s prosečnom starošću od 50 godina. U grupi pacijenata operisanih zbog folikulskog karcinoma (35 muških, 131 ţenska osoba) starosna dob je bila 15-78 godina, a prosečna starost 48 godina. Univarijantnom analizom pokazano je da se koncentracija tireoglobulina≥500 ng/ml, promer tumora<30 mm, prisustvo više od jednog tireoidnog čvora i nalaz afunkcijskog/hipofunkcijskog čvora značajno češće nalaze kod folikulskog karcinoma u odnosu na folikulski adenom. Nezavisni prediktivni faktori maligniteta bili su povišena preoperativna koncentracija tireoglobulina≥500 ng/ml i prisustvo više od jednog čvora. Ovi rezultati su, u cilju primene u praksi, prikazani i nomogramom, dvodimenzionalnim dijagramom dizajniranim da omogući pribliţno preoperativno grafičko izračunavanje verovatnoće postojanja maligniteta kod pacijenata s dijagnozom folikulske lezije nakon aspiracione biopsije tankom iglom. Zaključak: Povišena preoperativna koncentracija tireoglobulina≥500 ng/ml i prisustvo više od jednog čvora su nezavisni prediktori maligniteta folikulskih karcinoma štitaste ţlezde.

Establishing a preoperative diagnosis of thyroid follicular tumors is difficult due to the fact that the cell morphology of adenomas and carcinomas are similar and that capsular and vascular invasion cannot be determined by cytology. We analyzed predictive factors of biological behavior i.e. follicular carcinoma in order to enable a surgeon to indicate operative treatment and to perform an adequate operation for each patient with a follicular neoplasm. Method: In this retrospective study, we analyzed medical records of all patients with follicular thyroid tumors operated at Center for Endocrine Surgery, Clinical Center of Serbia, Belgrade between 2008 and 2012. A total of 263 operated patients were included and divided into follicular adenomas (97) and follicular carcinomas (166) based on the histopathology results. The most important demographic and clinical characteristics were analyzed by univariate and multivariate logistic regression analysis. Than, based on our results we formed a nomogram, a two-dimensional diagram designed to enable estimation of preoperative probability of malignancy in patients with diagnosis of the thyroid follicular lesion. Results: In adenoma group (19 males, 78 females) age range was 19-79, mean age 50. In carcinoma group (35 males, 131 females) age range was 15-78, mean age 48. Univariate analysis showed that thyroglobulin concentration≥500 ng/ml, tumor diameter<30 mm, presence of more than one thyroid nodule and an afunctional/hypofunctional nodule were significantly more frequent in follicular carcinoma than in follicular adenoma. Independent predictive factors of malignancy were elevated preoperative thyroglobulin concentration≥500 ng/ml and presence of more than one nodule. Based on our results we formed a nomogram, a two-dimensional diagram designed to enable estimation of preoperative probability of malignancy in patients with diagnosis of the thyroid follicular lesion by fine needle aspiration biopsy. Conclusion: Elevated preoperative thyroglobulin concentration≥500 ng/ml and the presence of more than one nodule are independent predictors of malignancy for follicular thyroid carcinomas.

Metodologija optimalnog projektovanja statorskog namotaja šestofaznih mašina uz uvažavanje frekvencijske zavisnosti parametara i magnetskog zasićenja

Optimal design of the stator winding in six-phase ac machines involving frequency dependent parameters and magnetic saturation

Upotreba električnih mašina u električnim vozilima i u elektromotornim pogonima velikih snaga često se bazira na konceptu višefaznih mašina i pretvarača. Predmet naučne rasprave u okviru disertacije je razmatranje, utvrđivanje i uvažavanje elektromagnetskih pojava koje se javljaju u toku rada ovih mašina, a koje mogu uticati na odabir optimalne topologije statorskog namotaja. Postoji potreba za analizom pojava koje se javljaju pri radu višefazne mašine, a koje utiču na promene njenih parametara i performansi, što je iskorišćeno kao temelj za formiranje metodologije projektovanja statorskog namotaja šestofazne mašine. Dok su adekvatni matematički modeli višefaznih mašina za linearne uslove rada dostupni, to nije slučaj sa modelima kod kojih je prisutno zasićenje magnetskog kola. U ovoj disertaciji obrađena je problematika projektovanja namotaja statora na primeru šestofazne mašine. Analizirana je pojava zasićenja magnetskog kola pri različitim uslovima napajanja, pri čemu se analizira potencijalna upotreba postojećih modela zasićenih trofaznih mašina za potrebe modelovanja zasićenih višefaznih mašina. Kod modela šestofazne mašine sa vektorski raspregnutim ravnima analizirano je postojanje međusobnog uticaja između različitih ravni pri pojavi zasićenja magnetskog kola, a što je od značaja za razvoj preciznijih modela i algoritama upravljanja. Disertacija se oslanja na analitička razmatranja, analizu primenom metode konačnih elemenata i eksperimentalne rezultate. Pokazano je da na zasićenje na putu glavnog fluksa utiču komponente struje koje ne doprinose stvaranju elektromehaničkog momenta, tj. ustanovljeno je postojanje međuravanskog unakrsnog zasićenja, što dovodi do potrebe za razvojem novih modela šestofaznih i ostalih višefaznih mašina, koji će uvažiti ovaj efekat. Pored zasićenja, analizirane su različite topologije statorskih namotaja radi ostvarivanja boljih performansi vi Doktorska disertacija | Miloš Ječmenica Univerzitet u Beogradu, Elektrotehnički fakultet šestofazne mašine sa stanovišta smanjenja površinskih gubitaka u rotoru. Ustanovljeno je da skraćenje navojnog koraka namotaja često ne dovodi do smanjenja ovih gubitaka. Kao rezultat analize formirana je metodologija optimalnog projektovanja statorskog namotaja šestofazne mašine koja se bazira na osnovnim naučnim doprinosima ove disertacije. Bližu oblast istraživanja predstavlja analiza pojava od kojih zavisi pristup projektovanju i optimizaciji statorskih namotaja naizmeničnih šestofaznih mašina. U disertaciji su opisane pojave koje se javljaju u karakterističnim radnim režimima, a od značaja su prilikom procesa projektovanja. Pri tome, najviše je pažnje posvećeno analizi efekta zasićenja magnetskog kola šestofazne mašine, kao i analizi gubitaka usled višeharmoničnih komponenti struja. Na osnovu sprovedene analize, u disertaciji je predložen analitički model koji uvažava efekte koji se javljaju pri zasićenju magnetskog kola šestofazne mašine. Predložena je nova ekvivalentna zamenska šema magnetskog kola statora, koja detaljnije predstavlja prirodu pojava u šestofaznoj mašini pri nastanku zasićenja. Ove pojave je potrebno uvažiti pri projektovanju same šestofazne mašine, kao i prilikom izrade upravljačkog algoritma. Pojava zasićenja magnetskog kola analizirana je analitičkim modelom, kao i metodom konačnih elemenata, nakon čega je na prototipu šestofazne mašine izvršena eksperimentalna verifikacija dobijenih rezultata. Mašina je ispitana pri napajanju iz šestofaznog pretvarača u režimima rada pri kojima nije izraženo zasićenje, kao i pri režimima rada sa pojavom zasićenja magnetskog kola. Eksperimentalni rezultati su pokazali dobro slaganje sa rezultatima analitičkog modela i rezultatima dobijenim metodom konačnih elemenata. Na ovaj način izvršena je verifikacija predloženog modela magnetskog kola šestofazne mašine, pri čemu je pokazano da prilikom zasićenja magnetskog kola dolazi do međusobnog uticaja komponenti struja koje ne utiču direktno u elektromehaničkom pretvaranju energije, sa onim komponentama koje pri tom slučaju prisutne. S tim u vezi, identifikovana je nova pojava koja se odnosi na spregu između međusobno ortogonalnih ravni ‒ pojava međuravanskog unakrsnog zasićenja. Postojanje ove pojave je potvrđeno analitički, metodom konačnih vii Doktorska disertacija | Miloš Ječmenica Univerzitet u Beogradu, Elektrotehnički fakultet elemenata i eksperimentalno. Iz dobijenih rezultata proizilazi potreba za uvažavanjem ove pojave prilikom projektovanja, naročito u pogledu optimizacije statorskog namotaja šestofaznih mašina, pri čemu je efekat sprege između ortogonalnih ravni značajan parametar koji utiče na izbor optimalne topologije namotaja. U nastavku istraživanja ispitan je uticaj frekvencijske zavisnosti parametara na projektovanje statorskog namotaja. Konstatovano je da način formiranja statorskog namotaja šestofazne mašine značajno utiče na vrednost njenih parametara, a, samim tim, i na njihovu frekvencijsku zavisnost. Analizirane su različite topologije namotaja statora šestofazne mašine, sa aspekta vrednosti indukovanih površinskih gubitaka u rotoru. Prikazana je razvijena metoda za proračun ovih gubitaka, kao i njene mogućnosti. Na osnovu dobijenih rezultata metodom konačnih elemenata, donete su preporuke od značaja za formiranje namotaja statora sa stanovišta redukovanja ovih gubitaka. Na osnovu prethodnog, definisana je metodologija projektovanja šestofaznih mašina, koja uvažava zaključke iz disertacije. Naučni doprinosi disertacije predstavljaju osnovu za razvoj novih i modifikaciju postojećih modela šestofaznih mašina. Ovi modeli bi u potpunosti uzeli u razmatranje pojavu zasićenja magnetskog kola i indukovane površinske gubitke u rotoru, prilikom projektovanja i optimizacije šestofaznih mašina i upravljanja njima. Prikazanim istraživanjima i dobijenim rezultatima u disertaciji otvoreno je nekoliko pravaca za dalje istraživanje u oblasti projektovanja, modelovanja i proračuna parametara šestofaznih mašina.

The use of electrical machines in electric vehicles and high-power drives frequently requires multiphase machines and multiphase inverters. The topic of the scientific discussion within the dissertation is the consideration, determination and appreciation of the electromagnetic phenomena that occur during the operation of multiphase machines, which can be related to the selection of the optimal topology of the stator winding. There is a need to analyze these phenomena, which influence the changes in parameters and performances. While appropriate mathematical models under the linear magnetic conditions are readily available for multiphase machines, the same cannot be said for the models of the saturated multiphase machines. This dissertation examines the saturation in an asymmetrical six-phase induction machine under different supply conditions and the possibility of extending the existing linear models to the saturated case. Specifically, in the vector space decomposed model, the mutual coupling between sequence planes is analyzed under saturated conditions, which is important for modeling and control purposes. The dissertation relies on analytical considerations, finite element analysis and experimental results. It is shown that the saturation of the main flux path is influenced by the current components in the orthogonal (non-fundamental) sequence plane. The existence of the cross-coupling effects between different subspaces is recognized, thus implies the need to develop new multiphase machine models which take this effect into account. In addition, various stator winding topologies have been analyzed in order to achieve better performance of the six-phase AC machine. An analysis of the different stator winding topologies and their influence on harmonic losses in the rotor cage, was presented. It has been found that the shortening of the winding pitch often has no effect on reducing these losses. The methodology of designing the stator winding of a six-phase machine is presented, which takes into account the contributions presented in the dissertation. ix Докторска дисертација | Милош Јечменица Универзитет у Београду, Електротехнички факултет A closer study of the dissertation research is the analysis of the phenomena which are significant in the design and optimization procedure when constructing a six- phase AC machine. The dissertation describes the phenomena that occur during characteristic operating regimes and are of importance during the design process. Therein, the magnetic circuit saturation effect was analyzed in greatest detail, while considering the existing methods for modeling in both three-phase and six-phase machines. It was concluded that the frequency dependence of parameters is present as in three-phase machines. An analytical model that describes the saturation effect in the six-phase machine is proposed. Also, effects caused by the frequency variation from the stator winding design standpoint are also analyzed. The effect of frequency dependency of parameters is present as in three-phase machines, but from the point of view of modeling and control, it can often be ignored. The proposed equivalent magnetic circuit model provides an insight into the saturation phenomenon in a six-phase machine. The saturation-related phenomena have to be considered in the machine design process, as well as for the control purposes. In addition to analytical considerations, the saturation phenomenon was analyzed in more detail using the finite element method, after which the experimental verification of the obtained results was carried out on a dedicated six- phase machine. The machine was tested with a six-phase converter supply in operating modes with low saturation, as well as in operating modes with pronounced saturation. The experimental results showed a good match with the results of the analytical model and the results obtained by the finite element method. In this way, the verification of the proposed saturated analytical model of the six-phase machine was performed, showing that the saturation of the main flux path is influenced not only by the torque-producing current components, but also by the orthogonal current components that are not related to electromechanical energy conversion. Consequently, a phenomenon that relates to the cross-coupling effects between different subspaces in the vector space decomposed model of the six-phase machines has been identified as interplane cross-saturation. The procedures carried out in this research have confirmed analytically, by means of finite element method and experimentally the need to acknowledge the discovered effect during the design process, most notably in the design process of the stator winding of x Докторска дисертација | Милош Јечменица Универзитет у Београду, Електротехнички факултет six-phase machines. Additionally, the coupling effect between orthogonal subspaces is significant from the control standpoint. In addition of the research, the influence of the frequency dependence of the parameters on the stator winding topology was examined. It was concluded that the design of the stator winding of the six-phase machine significantly influences the value of its parameters, and consequently also on their frequency dependence. Various topologies of the stator windings from the aspect of the induced harmonic losses in the rotor, was analyzed. The developed method for calculating these losses as well as its possibilities is presented. On the basis of the obtained results with the finite element method, recommendations were made to form optimal stator winding topology. Based on this, the methodology for designing six-phase machines has been defined, which introduces into consideration the impact of this newly identified phenomena and conclusions from this dissertation. This is intended to motivate the development of new and modification of existing models of six-phase machines so that the saturation effect and the aspect of the induction of harmonic losses in the rotor will be properly considered in the design, optimization and control of six-phase machines. With the presented research and obtained results in the dissertation, the entire research area in the field of design, modeling and calculation of parameters of six-phase machines has been opened.

Dekorativno oblikovanje tradicionalne narodne arhitekture u regionu Stare planine

Decorative forms of traditional vernacular architecture in Stara planina region.

regionu Stare planine, Republika Srbija, u 19. i početkom 20. veka stihijski se stvarao fond narodnog graditeljskog nasleđa, što danas predstavlja kulturno-umetničko bogatstvo naroda Srbije. Dekorativno oblikovanje, kao čovekova potreba da ulepša i na njemu prepoznatljiv način obeleži prostor u kome živi, iznedrilo je lepotu oblika i u ovim krajevima. U doktorskoj disertaciji predstavljeni su i analizirani karakteristični dekorativni oblici i elementi tradicionalne arhitekture u regionu Stare planine, posmatrano kroz njihove kulturne, istorijske, etnografske i umetničko-zanatske vrednosti. Disertacija je formirana na osnovu građe prikupljene na terenu u proteklih pet godina, obilaskom, snimanjem i analizom arhitekture 37 planinskih sela Pirotskog okruga, Pirota i Dimitrovgrada. Detaljnom analitičkom metodom sprovedena je analiza autentičnosti oblika, a tipološko-komparativnom metodom najpre su klasifikovani i definisani osnovni tipovi dekorativnih oblika prema formi, poreklu i nastanku, tehnici građenja i materijalu, pa zatim komparirani sa karakterističnim primerima dekoracija u širem regionu staroplanske oblasti (Srbije i Bugarske). Tradicionalnu arhitekturu čine: konstruktivni sistemi, ispune, krovne strehe, krovne konstrukcije, jastuci, stubovi, tremovi, doksati, način i oblik zidanja dimnjaka i ognjišta, ćenarke, čuvarkuće itd., tehnikama gradnje prilagođeni prirodnim materijalima, i oni predstavljaju autentično i neponovljivo nasleđe koje determinišu istorijska komponenta, kvalitet i duh vremena stroplaninskog naroda...

In the period of the 19th century and the beginning of the 20th century, a number of vernacular architectural structures, nowadays regarded as cultural and artistic heritage of Serbian people, were spontaneously built in the region of Stara Planina mountain. Owing to decoration, as an expression of people’s need to embellish and, in a unique manner, enrich the space they live in, the mentioned area possesses a wealth of beautiful forms and elements. This doctoral dissertation is aimed at presenting and analyzing the characteristic decorative forms and elements of the traditional architecture in the Stara Planina region, with an emphasis on their cultural, historical, ethnographical and arts-and-crafts values. The material for the dissertation has been collected in the past 5 years, by visiting the mentioned region and by surveying and analyzing the architecture in 37 mountain villages in the areas of Pirot District and the cities of Pirot and Dimitrovgrad. By the detailed analytical method, the analysis of the authentic forms has been carried out, whereas the application of the comparative method has enabled classification and definition of basic types of decorative forms according to their form, origin and formation, construction techniques and material, as well as their subsequent comparison with the characteristic decorative forms in the broader surroundings of the Stara Planina region in Serbia and Bulgaria. Authentic and unique tradition, which reflects the historical component of the life quality and the spirit of the past times, has been preserved in the elements of traditional architecture in the Stara Planina villages. This architecture is presented by: traditional construction systems, infills, roof eaves, roof structures, bolsters, pillars, porches, porches known as doksati, methods and types of the construction of chimneys and fireplaces, ćenarke, čuvarkuće etc., with the construction techniques adjusted to natural materials...

Uticaj citrusnih flavanona na tireoidnu homeostazu i lipidni status starih pacova

Impact of citrus flavanones on thyroid homeostasis and lipid status of old-aged rats

Citrusni flavanoni, naringenin (NAR) i hesperetin (HES), su polifenolne supstance koje zbog sterne sličnosti sa endogenim tiroksinom (T4) mogu da interferiraju sa aktivnošću tiroidne peroksidaze, ključnog enzima u biosintezi tiroidnih hormona. Sa starenjem se povećava osetljivost tiroidne osovine na efekte egzogenih supstanci i raste rizik od nastanka hroničnih bolesti usled opadanja detoksikacione sposobnosti i akumulacije lipida u jetri. Cilj ove doktorske disertacije bio je da se ispita uticaj NAR i HES na tiroidnu homeostazu i lipidni status 24 meseca starih mužjaka pacova Wistar soja. Supstance su aplikovane per os u dozi od 15 mg/kg t.m., tokom četiri nedelje. Rezultati ove disertacije su pokazali da NAR i HES uzrokuju strukturne i ultrastrukturne promene štitaste žlezde starih mužjaka pacova, povećavaju imunohistohemijsku ekspresiju tiroglobulina za koji je vezan T4 i vaskularnog endotelijalnog faktora rasta. Ekspresija trijodtironinom (T3) regulisanih gena u jetri je snižena nakon tretmana citrusnim flavanonima. U serumu, NAR je povisio koncentracije TSH i HDL, dok je koncentracija antioksidativnih enzima jetre snižena nakon oba tretmana. Uočava se desaturacija masnih kiselina u fosfolipidima membrane hepatocita, povećanje udela n-3, a sniženja zastupljenosti n-6 masnih kiselina nakon primene oba citrusna flavanona. Smanjuje se lipidna peroksidacija membrane hepatocita i poboljšava antioksidativna zaštita u jetri nakon NAR i HES tretmana. NAR je ispolji jači efekat, jer je uticao na aktivnost, gensku i proteinsku ekspresiju antioksidativnih enzima, i parametre redoks ravnoteže, dok je HES pojačao antioksidativnu zaštitu, ali u manjoj meri. Upotreba citrusnih flavanona uzrokuje neželjene efekte - subkličničku hipotireozu koju prate povišen nivo TSH i smanjena ekspresija T3 regulisanih gena u jetri. Međutim, sa stanovišta desaturacije membrane hepatocita i poboljšanja antioksidativne zaštite u jetri, upotreba NAR i HES se može smatrati korisnom, imajući u vidu uticaj ovih promena na produženje dužine životnog veka.

Citrus flavanones, naringenin (NAR) and hesperetin (HES), are polyphenolic substances, structurally similar to endogenous thyroxine (T4), that can interfere with the activity of thyroid peroxidase, the key enzyme in thyroid hormone biosynthesis. With aging, thyroid axis sensitivity to the effects of exogenously applied substances increases and the risk of chronic diseases raises due to reduced detoxification ability and the accumulation of lipids in the liver. The aim of this doctoral dissertation was to examine the impact of NAR and HES on thyroid homeostasis and the lipid status of 24 months- old Wistar male rats. The substances were administered per os at a dose of 15 mg/kg b.w., for four weeks. The results of this dissertation have shown that NAR and HES cause structural and ultrastructural changes in the thyroid gland of old male rats, increasing the immunohistochemical expression of T4 bounded to thyroglobulin as well as of vascular endothelial growth factor. Expression of triiodothyronine (T3) regulated genes in the liver were decreased after treatment with citrus flavanones. In the serum, the NAR increased concentrations of TSH and HDL, while the concentration of liver antioxidative enzymes was reduced after both treatments. The desaturation of fatty acids in the hepatocyte membrane phospholipids, the increase in the proportion of n-3, and the reduction in the presence of n-6 fatty acids after application of both citrus flavanones is observed. Lipid peroxidation of the hepatocyte membrane is reduced and the antioxidant protection is improved in the liver after NAR and HES treatments. The NAR has a stronger effect, as it has affected the activity, gene and protein expression of antioxidant enzymes, as well as the parameters of redox balance, while HES enhanced antioxidant protection, but to a less degree. The use of citrus flavanones causes unwanted effects - subclinical hypothyrosis followed by elevated TSH and reduced expression of T3 regulated genes in the liver. However, from the perspective of the hepatocyte membrane desaturation and the enhancement of antioxidant protection in the liver, the use of NAR and HES can be considered useful, given that the effects of these changes are associated with the extend life span.

Efekti interleukina-17 na funkcionalna svojstva humanih mezenhimskih matičnih ćelija periodoncijuma

Effects of interleukin-17 on functional properties of human periodontal ligament mesenchymal stem cells

Istraživanja regenerativnih i modulatornih uloga mezenhimskih matičnih ćelija (MMĆ) u poslednjih nekoliko decenija su zaokupirala pažnju naučnika s obzirom da ove ćelije ispoljavaju karakteristike koje ukazuju na mogućnosti njihove primene u regenerativnoj medicini, kao i na polju ćelijske terapije. Iako MMĆ izolovane iz različitih adultnih i neonatalnih izvora ispoljavaju potencijal samoobnove i multipotentne diferencijacije (u ćelije mezodermalnog, ali i ektodermalnog i endodermalnog porekla), mnoga istraživanja koja se tiču funkcionalnih karakteristika ovih ćelija tek predstoje kako bi njihova primena bila efikasna i bezbedna. Jedan od novije otkrivenih izvora MMĆ je i tkivo periodoncijuma koje predstavlja bogat i lako dostupan izvor MMĆ, s obzirom da se ovo tkivo dobija nakon ekstrakcije zuba iz ortodontskih razloga. Periodoncijum je vezivno tkivo čija je glavna uloga uspostavljanje čvrste veze između korena zuba i alveolarne kosti. Regenerativni potencijal MMĆ periodoncijuma (PD-MMĆ) ogleda se u njihovom potencijalu samoobnove i multipotentne diferencijacije, kao i sposobnosti formiranja tkiva sličnih potpornom tkivu zuba, uključujući periodoncijum i cement. Ove osobine čine PD-MMĆ dobrim kandidatima za primenu u regeneraciji potpornog tkiva zuba u parodontopatiji, hroničnom inflamatornom oboljenju uzrokovanom bakterijama zubnog plaka. Razvoj hronične parodontopatije podrazumeva procese remodelovanja koštanog tkiva pod uticajem brojnih faktora u koje su uključeni osteoblasti i osteoklasti koji ostvaruju recipročne interakcije sa ćelijama imunskog odgovora regulišući inflamaciju i degradaciju koštanog tkiva. Uloga lokalne mikrosredine u parodontopatiji predstavlja jedan od ključnih faktora u kontekstu ćelijskog odgovora tkiva periodoncijuma na perzistentnu infekciju. U okviru parodontalnih lezija potvrđeno je prisustvo Th17 (od engl. T helper cells17) ćelija, značajnih regulatora tkivnog razaranja u toku zapaljenskih procesa, koje produkuju proinflamatorni citokin interleukin (IL)-17. U kontekstu parodontopatije, do sada je pokazano da IL-17 ostvaruje svoju ulogu stimulišući privlačenje neutrofila na mesto inflamacije kao i direktnim stimulatornim delovanjem Th17 ćelija na diferencijaciju osteoklasta. S druge strane, poznato je da IL-17 reguliše funkcionalne osobine, poput proliferacije i diferencijacije, kako mišjih tako i humanih MMĆ pri čemu je pokazano da ispoljeni efekti IL-17 zavise od vrste davaoca i tkivnog porekla MMĆ. Kako u literaturi nema dovoljno podataka o biološkim efektima i mehanizmima delovanja IL-17 na funkcionalna svojstva PD-MMĆ, ciljevi ove disertacije su, pored izolacije i karakterizacije PD-MMĆ, bili usmereni na ispitivanje uticaja IL-17 na funkcionalna svojstva ovih ćelija kao i molekularnih mehanizama uključenih u delovanje IL-17 na PD- MMĆ. Kako bi se zadovoljili kriterijumi Komiteta za mezenhimske matične ćelije Međunarodnog društva za ćelijsku terapiju, karakterizacija PD-MMĆ izolovanih iz zdravih osoba je izvedena ispitivanjem sposobnosti adhezije izolovanih ćelija za plastiku, određivanjem ekspresije markera mezenhimskih, hematopoetskih, kao i embrionalnih matičnih ćelija kao i ispitivanjem sposobnosti PD-MMĆ da se diferenciraju u tri ćelijske loze (osteogenu, adipogenu i hondrogenu) nakon kultivacije u odgovarajućim diferencijacionim medijumima. U drugom delu istraživanja ispitivani su uticaji i mehanizmi delovanja IL-17 na funkcionalna svojstva PD-MMĆ, uključujući njihovu proliferaciju, klonogeni kapacitet, migraciju i osteogenu diferencijaciju. U kontekstu osteogene diferencijacije praćena je ekspresija gena za markere osteogeneze (alkalnu fosfatazu (ALP), Osteoklacin (Ocn) i RUNX2/Cbfa1 (od engl. Runt-related transcription factor 2/Core binding factor 1)) u PD- MMĆ kultivisanim u osteogenom diferencijacionom medijumu. Dodatno je ispitivano učešće MAPK (od engl. Mitogen Associated Protein Kinases) signalnih puteva u delovanju IL-17 na funkcionalne osobine PD-MMĆ. Trećim segmentom istraživanja obuhvaćena su ispitivanja uticaja i mehanizama delovanja IL-17 na ekspresiju i aktivnost enzima uključenih u razgradnju komponenti ekstraćelijskog matriksa, poput matriksnih metaloproteinaza (MMP) i urokinaze (uPA, od engl. urokinase type plasminogen activator) u PD-MMĆ. Uz to, ispitivano je i učešće MAPK signalnih puteva u delovanju IL-17 na ekspresiju i aktivnost ovih enzima u PD-MMĆ. U poslednjem segmentu istraživanja ispitivan je uticaj IL-17 na imunomodulatorne osobine PD-MMĆ određivanjem delovanja IL-17 na potencijal PD-MMĆ da modulišu spontanu i mitogenom-stimulisanu proliferaciju T limfocita periferne krvi, kao i analizom njegovih efekata na gensku ekspresiju molekula povezanih sa imunomodulatornim delovanjem PD-MMĆ, uključujući HLA (od engl. Human Leukocyte Antigen)-A, HLA-DR, HLA-G, IL-6 i indoleamindioksigenaza-1 (IDO-1). Rezultati dobijeni u sklopu ove disertacije su pokazali da su PD-MMĆ uspešno izolovane iz svih ispitivanih uzoraka pri čemu je utvrđeno da u skladu sa kriterijumima Međunarodnog društva za ćelijsku terapiju izolovane ćelije ispoljavaju osnovne karakteristike MMĆ uključujući adherentnost za plastiku, ekspresiju specifičnih markera MMĆ, uz odsustvo ekspresije markera hematopoetskih ćelija, kao i multipotentni potencijal diferencijacije u ćelije osteogene, hondrogene i adipogene loze. Uz to, naši rezultati su pokazali i da izolovane PD-MMĆ eksprimiraju markere embrionalnih matičnih ćelija, Nanog, Oct4, SOX-2 i SSEA4, kao i da ispoljavaju receptor za IL-17. Ispitivanja uticaja IL-17 na funkcionalna svojstva PD-MMĆ su pokazala da IL-17 dovodi do blagog smanjenja proliferacije ovih ćelija samo nakon 24h inkubacije, dok s druge strane stimuliše klonogeni kapacitet PD-MMĆ indukujući povećanje broja CFU-F (od engl. Colony Forming Unit Fibroblast) kolonija sličnih fibroblastima. Takođe je utvrđeno da IL-17 inhibira migraciju PD-MMĆ, kao i osteogenu diferencijaciju ovih ćelija indukujući smanjenje ekspresije gena za ALP, RUNX2/Cbfa1 i Ocn. Uz to je pokazano da IL-17 aktivira sve tri klase MAPK: p38, ERK1,2 i JNK u PD-MMĆ, kao i da se inhibitorni efekat IL-17 na osteogenu diferencijaciju PD-MMĆ ostvaruje preko aktivacije ERK1,2 i/ili JNK MAPK signalnih puteva. Rezultati dobijeni u sklopu narednog segmenta istraživanja su pokazali da IL-17 indukuje povećanu enzimsku aktivnost, proteinsku i gensku ekspresiju MMP-2 i uPA u PD-MMĆ, dok nema efekta na ekspresiju MMP-9 u ovim ćelijama. Dodatno su naši nalazi pokazali da je stimulatorni efekat IL-17 na ekspresiju MMP-2 i uPA posredovan aktivacijom ERK1,2 MAPK signalnog puta. U poslednjem segmentu naših ispitivanja uticaja IL-17 na imunomodulatorne osobine PD- MMĆ dobijeni rezultati su pokazali da IL-17 nema uticaja na sposobnost PD-MMĆ da suprimiraju proliferaciju mitogenom stimulisanih T limfocita. Analizom genske ekspresije molekula povezanih sa imunomodulatornim delovanjem PD-MMĆ utvrđeno je da IL-17 nema uticaja na ekspresiju HLA-A gena, dok dovodi do smanjenja genske ekspresije HLA- G5 u PD-MMĆ. Takođe je pokazano da IL-17 indukuje povećanje genske ekspresije HLA- DR u PD-MMĆ, ali da nema uticaja na površinsku ekspresiju ovog molekula. Uz to, pokazan je stimulatorni efekat IL-17 na ekspresiju gena za IDO-1 i IL-6 u PD-MMĆ. Imajući u vidu predstavljene rezultate, može se zaključiti da PD-MMĆ predstavljaju dobar izvor MMĆ koje zadovoljavaju sve kriterijume za karakterizaciju ovih ćelija. Ispitivanja uticaja IL-17 na funkcionalne osobine PD-MMĆ su pokazala da IL-17 inhibira migraciju i osteogenu diferencijaciju PD-MMĆ, pri čemu efekte na osteogenu diferencijaciju ostvaruje putem aktivacije ERK1,2 i JNK MAPK. U kontekstu ispitivanja uticaja IL-17 na produkciju enzima uključenih u razgradnju ekstraćelijskog matriksa, utvrđeno je da IL-17 indukuje ekspresiju enzimski aktivne MMP-2 i uPA putem aktivacije ERK1,2 MAPK. Dobijeni podaci ispitivanja uticaja IL-17 na fukcionalna svojstva PD-MMĆ ukazuju na potencijalne mehanizme delovanja IL-17 u razvoju paradontopatije i ističu kompleksnost biologije MMĆ i njihove interakcije sa faktorima mikrosredine postavljajući osnovu budućim istraživanjima u cilju pronalaženja novih modaliteta lečenja ovog široko rasprostranjenog inflamatornog oboljenja.

In the last few decades regenerative and modulatory properties of mesenchymal stem cells (MSCs) have gained the attention of scientists since their characteristics indicated the possibility of their use in regenerative medicine and cell therapy. Although MSCs isolated from various adult and neonatal tissues exhibit the potential of self-renewal and multipotent differentiation (towards cells of mesodermal, ectodermal and endodermal origin), many aspects concerning their functional characteristics are yet to be explored. One of the recently discovered sources of MSCs is easily accessible tissue of periodontal ligament, obtained after tooth extraction for orthodontic reasons. Periodontal ligament tissue strengthens the root of the tooth in the alveolar bone. The regenerative potential of periodontal ligament MSCs (PDLSCs) is reflected in self-renewal and multipotent differentiation ability, as well as potency for the formation of tissues which support the teeth, including the periodontal ligament and cement. These features make the PDLSCs good candidates for use in the regeneration/reconstruction of supporting tissue of teeth in periodontal disease, a chronic inflammatory disease caused by bacterial plaque. The development of chronic periodontal disease includes the process of bone tissue remodeling governed by a number of factors, including osteoblasts and osteoclasts, which reciprocally interact with cellular and humoral compartments of the immune system, thus regulating inflammation, degradation and renewal of bone tissue. The role of the local microenvironment in the periodontal disease is one of the key factors in the context of periodontal ligament tissue response to persistent infection. The presence of T helper cells17 (Th17) cells, important regulators of tissue destruction during inflammatory processes has been confirmed in periodontal lesions. Th17 cells produce proinflammatory cytokine interleukin (IL)-17. In the context of periodontal disease, as well as in other inflammatory diseases, IL-17 stimulates the activation and migration of neutrophils to the site of inflammation. Additionally, IL-17 directly stimulates differentiation of osteoclasts and contributes to the alveolar bone resorption. On the other hand, it is known that IL-17 regulates functional properties, such as proliferation and differentiation, of both murine and human MSCs, in a manner that depends on the type of host and tissue origin of MSCs. Since there is not enough data about biological effects of IL-17 on functional properties of PDLSCs and mechanisms involved, the objectives of this thesis include the isolation and characterization of PDLSCs, the analyses of IL-17 effects on functional properties of these cells, as well as investigation of IL-17-initiated underlying mechanisms involved in regulation of PDLSCs properties. In order to meet the criteria of the Mesenchymal and Tissue Stem Cell Committee of the International Society for Cellular Therapy, characterization of PDLSCs isolated from healthy individuals has been performed by testing the ability of isolated cells to adhere to plastic surface, determining the expression of mesenchymal, hematopoietic, and embryonic stem cells markers as well as the potential of PDLSCs to differentiate into three cell lineages (osteogenic, adipogenic and chondrogenic) after cultivation in appropriate differentiation medium. In the second part of the study, we have examined the effects of IL-17 on functional properties of PDLSCs, including their proliferation, clonogenic capacity, migration and osteogenic differentiation. In the context of osteogenic differentiation, we have examined gene expression of markers for osteogenesis (alkaline phosphatase (ALP), Osteocalcin (Ocn), Runt-related transcription factor 2 / Core binding factor 1 (RUNX2 / Cbfa1) in the PDLSCs cultured in osteogenic medium. In addition, we have investigated the participation of Mitogen Associated Protein Kinases (MAPK) signaling pathways in the IL-17-mediated effects on functional properties of PDLSCs. The third segment of the research has included the analyses of the IL-17 influence and its mechanisms of action on the expression and activity of enzymes involved in the degradation of extracellular matrix components, such are matrix metalloproteinases (MMP) and urokinase type plasminogen activator (uPA) in PDLSCs. In addition, we have investigated the involvement of MAPK signaling pathways in the IL-17-mediated expression of these enzymes in PDLSCs. In the last segment of our study, we have examined the effects of IL-17 on the immunomodulatory properties of PDLSCs by determining the potential of IL- 17 to modulate effects of PDLSCs on spontaneous and mitogen-stimulated proliferation of peripheral blood T lymphocytes. Moreover, the effects of IL-17 on gene expression of molecules associated with PDLSCs immunomodulatory properties, such as Human Leukocyte Antigen (HLA)-A, HLA-DR, HLA-G5, IL-6 and Indoleamine 2,3-dioxygenase-1 (IDO-1) have been analyzed. Results obtained in this thesis showed that PDLSCs have been successfully isolated from all human samples and that morphological and phenotypic characteristics of isolated cells are in agreement with the criteria of the International Society for Cellular Therapy. Isolated cells exhibited the basic characteristics of MSCs including adherence to plastic, the expression of specific MSCs markers (with absence of hematopoietic cells markers expression) and multipotent differentiation potential into cells of osteogenic, chondrogenic and adipogenic lineages. In addition, our results have shown that the isolated PDLSCs express markers of embryonic stem cells, Nanog, Oct4, SOX-2 and SSEA4, as well as the receptor for IL-17. Investigation of the effects of IL-17 on the functional properties of PDLSCs have shown that IL-17 leads to a slight decrease in the proliferation of these cells only after 24 hours of incubation, while it stimulates the clonogenic capacity of PDLSCs increasing the number of Colony Fibroblast forming Unit (CFU-F) colonies. Also, we have found that IL-17 inhibits the migration of PDLSCs and the osteogenic differentiation of these cells, reducing gene expression for ALP, RUNX2 / Cbfa1 and Ocn. In addition, we have shown that IL-17 activates all three classes of MAPK: p38, JNK and ERK1,2 in PDLSCs, and that the inhibitory effect of IL-17 on the osteogenic differentiation of PDLSCs is achieved through activation of ERK1,2 and /or JNK MAPK signaling pathways. The results obtained in the next segment of research have shown that IL-17 induces enzymatic activity, the protein and the gene expression of MMP-2 and uPA in the PDLSCs, while having no effect on the expression of MMP-9 in these cells. In addition, our findings have indicated that the stimulatory effect of IL-17 on the expression of MMP-2 and uPA is mediated via ERK1,2 MAPK signaling pathway activation. In the final segment of our investigation we have examined the effect of IL-17 on the immunomodulatory properties of PDLSCs. Obtained results have shown that IL-17 has no influence on the ability to PDLSCs to suppress the proliferation of mitogen-stimulated T lymphocytes. Analyses of gene expression of molecules associated with the immunomodulatory properties of the PDLSCs have shown that IL-17 has no effect on the expression of the HLA-Agene, while it decreases the gene expression of HLA-G in the PDLSCs. Also, it has been demonstrated that IL-17 induces an increase in gene expression of HLA-DR in the PDLSCs, while it has no impact on the surface expression of this molecule. In addition, our results have shown the stimulatory effect of IL-17 on the expression of IDO- 1 and IL-6 genes in the PDLSCs. Bearing in mind presented results, we can conclude that PDLSCs are a good source of MSCs that meet all the criteria for characterization of these cells. Analyses of the influence of IL-17 on the functional properties of PDLSCs have shown that IL-17 inhibits migration and osteogenic differentiation of PDLSCs, wherein the effects on osteogenic differentiation were achieved through activation of the ERK1,2 and JNK MAPKs. Investigations of the IL-17 effects on the production of enzymes involved in the degradation of extracellular matrix have demonstrated that IL-17 induces the expression of enzymatically active MMP-2 and uPA via ERK1,2 MAPK activation. Research data concerning the influence of IL-17 on the functional properties of PDLSCs point to the potential mechanisms of IL-17 action in the development of periodontal disease and highlight the complexity of MSCs biology and their interactions with the microenvironment factors laying the basis for future research aimed at finding the new modalities of treatment for this widespread inflammatory disease.

Ekspresija proteina održavanja minihromozoma u keratinocitnoj intraepidermalnoj neoplaziji i invazivnom planocelularnom karcinomu kože

Expression of minichromosome maintenance proteins in keratinocytic intraepidermal neoplasia and invasive squamous cell carcinoma of the skin

Aktinična keratoza (AK) predstavlja proliferaciju neoplastično transformisanih keratinocita ograničenih na deo epiderma i smatra se inicijalnom kliničkom manifestacijom bolesti koja može da progredira do planocelularnog tj. skvamocelularnog karcinoma (engl. squamous cell carcinoma, SCC), ranim stadijumom u biološkom spektru SCC. Iz tog raloga, a prema konceptu progresije u evoluciji AK, predložena je nova nomenklatura za AK, keratinocitna intraepidermalna neoplazija (engl. keratinocyte intraepidermal neoplasia, KIN), prema anlanogiji sa cervikalnom intraepidermalnom neoplazijom. Skvamocelularni karcinom kože in situ (engl. squamous cell carcinoma in situ - SCCIS) ili Bowenova bolest (engl. Bowen disease, BD) je maligna neoplazma ograničena na epiderm, kod koga se neoplastično tranformisani keratinociti nalaze čitavom debljinom epiderma, bez znakova dermalne invazije. Kutani SCC je drugi najčešći maligni tumor kože, koji se manifestuje različitim promenama od lako izlečivih, superficijalno invazivnih kancera do veoma infiltrovanih tumora koji metastaziraju dovodeći do smrtnog ishoda. Može nastati de novo, ali češće nastaje iz prekursorskih lezija, AK i SCCIS, koje, kao što je već pomenuto, prema pojedinim autorima predstavljaju SCC u ranijim evolutivnim fazama. Iako AK, SCCIS i SCC mozemo smatrati različitim stadijumima iste bolesti, ne progrediraju sve lezije u SCC. Molekularni mehanizmi ove progresije još uvek nisu u potpunosti razjašnjeni. Proteini održavanja minihromozoma (engl. minichromosome maintenance proteins, MCM) predstavljaju kompleks proteina koji ima ključnu ulogu u regulaciji replikacije DNK tokom ćelijskog ciklusa. MCM grupa jedarnih proteina učestvuje u započinjanju i održavanju replikacije DNK molekula. Usklađenost funkcionalne interakcije između MCM proteina i drugih komponenti, posredstvom protein kinaza koje učestvuju u regulaciji ćelijskog ciklusa, omogućava početak sinteze DNK molekula samo jednom tokom ćelijskog ciklusa, održavanjući, na taj način stabilnost genoma ćelije. MCM proteini eksprimovani su u ćelijama u kojima je ćelijski ciklus u toku, dok se u ćelijama koje su u stanju mirovanja i diferencijacije njihova ekspresija gubi, pa predstavljaju potencijalno korisne markere proliferacije ćelija, posebno u tumorima. Kancerogeneza planocelularnih (skvamocelularnih) neoplazmi kože još uvek nije u potpunosti razjašnjena i u fazi je ispitivanja. Jedan od poremećaja koji ima uticaja na neadekvatnu proliferaciju keratinocita svakako je i abnormalna ekspresija i funkcija MCM proteina. Cilj ove studije bio je ispitivanje ekspresije MCM 2, 5 i 7 u AK, BD i invazivnom SCC, analiza ispoljavanja ovih proteina u in situ lezijama klasifikovanim kao keratinocitna intraepidermalna neoplazija (KIN), poređenje ekspresije MCM 2, MCM5 i MCM7 i prognostičkih parametara SCC kao što su dimenzije, dubina invazije, debljina i gradus tumora, kao i procena različitih načina evaluacije imunohistohemijskog bojenja i ekspresije MCM proteina u odnosu na tip lezije i prognostičke paramete. Istraživanje je sprovedeno na 315 slučajeva, od čega 91 promena AK, 50 promena BD i 174 invazivni SCC, od toga podjednak broj dobro (G1), srednje (G2) i slabo (G3) diferentovanih SCC (po 58 slučajeva). Prema KIN klasifikaciji, od 141 preinvazivne lezije, 50 promena KIN 1, 40 KIN 2, a 51 KIN 3. Ekspresija MCM2, MCM5 i MCM7 proteina je ispitivana imunohistohemijski (IHH). Radi efikasnije, ekonomičnije, brže i lakše analize tokom IHH pocesa korišćena je tehnika tkivnog mikroniza, (engl. tissue microarray – TMA). Analizirani su distribucija imunopozitivnosti i broj pozitivnih ćelija tj. obim ekspresije (semikvantitativno). Ekspresija MCM2, MCM5 i MCM7 bila je značajnao različita kod svakog od pojedinačnih, ispitivanih histopatoloških entiteta. U analizi distribucije pozitivnih ćelija bazalna distribucija bila je dominatna kod AK, difuzna kod BD, dok su periferna i difuzna distrivucija bile gotovo podjednako zastupljene kod SCC. U slučaju in situ lezija, klasifikovanih kao KIN, difuzna distribucija bila je uočena ćešće kod KIN3 u odnosu na KIN1, KIN2 ili KIN1 i KIN2 zajedno. Takođe, difuzna distribucija bila je izraženija kod KIN2 lezija u odnsou na KIN1. U slučaju SCC, MCM2, MCM5 i MCM7 pokazala su dominantno perifernu distibuciju u najvećem broju slučajeva dobro diferentovanih SCC (G1). U slučaju srednje diferentovanih SCC (G2) periferna i difuzna distribucija sva tri proteina bile su uočene u gotovo podjednakom broju slučajeva, dok je kod najvećeg broja slabo diferentovanih SCC (G3) dominirala difuzna distribucija pozitivnih ćelija. Analizom distribucije MCM2, MCM5 i MCM7 u odnsou na prognostičke parametre SCC iako je kod većih i debljih tumora difuzna distribucija bila dominatna, značajna razlika uočena je u slučaju kada je granica debljine tumora iznosila 6mm. Semikavantitativnom procentom broja pozitivnih ćelija, sa granicom 50% utvrđena je razlika u obimu ekspresije u odnosu na histopatološki tip lezije. Procenat pozitivnih MCM2, MCM5 i MCM7 ćelija rastao je od AK, BD do SCC, ali je razlika u broju pozitivnih ćelija bila je značajna samo za MCM2 i MCM5. U slučaju MCM7, utvrđena je značajna razlika u ekspresiji između AK i SCC. I kod KIN lezija procenat pozitivnih ćelija rastao je od KIN1 do KIN3. Obim ekspresije MCM2, MCM5 i MCM7 bio je različit između KIN3, KIN1 i KIN2, kao i kada su KIN1 i KIN2 činile jednu grupu. Razlika u ekspresiji MCM2 i MCM5 bila je manja između KIN1 i KIN2, a u slučaju MCM7 i nije imala statističku značajnost. Ekspresija sva tri proteina takođe se razlikovala kod SCC različitog stepena diferentovanosti. Procenat MCM2, MCM5 i MCM7 pozitivnih ćelija povećavao se od dobro ka slabo diferentovanim tumorima i bio značajno različit između tumora različitih gradusa, SCC G1, SCC G2 i SCC G3. Pojedinačnim poređenjem, razlika u ekspresiji bila nešto manje izražena između SCC G2 i G3. Kada je analiziran obim ekspresije MCM2, MCM5 i MCM7 pozitivnih ćelija, sa granicom 50%, u odnosu na prognostičke parametre SCC, utvrđena je razlika u ekspresiji sva tri proteina samo u odnsou na debljinu tumora, kada je granica debljine iznosila 6mm. Ekspresija se nije značajno različikovala kod SCC različite veličine. Ovo je prva studija u kojoj je procenjivana distribucija i ekspresija MCM proteina u keratinocitnim neoplazmama kože. Prema rezulatima ove studije difuzna distribucija i veći obim ekspresije MCM2, MCM5 i MCM7 proteina ćelijskog cikusa u BD u odnosu na AK, KIN3 u odnosu na KIN1 i KIN2, kao i kod slabodiferentovanih SCC u odnosu na dobrodiferentovane SCC ukazuje na njihovu ulogu u progresiji in situ i invazivnih keratinocitnih neoplazmi. Utvrđena korelacija sa debljinom tumora 6mm, kao prognostičkim parametrom SCC, ukazuje na zaključak da je ekspresija MCM2, MCM5 i MCM7 proteina udružena sa faktorima rizika SCC. Takođe, činjenica da su slični rezultati dobijeni u slučaju svakog od proteina pojedinačno govori u prilog i dokazuje njihov značaj u evaluaciji progresije in situ lezija, kao i mogućih prognostičkih indikatora invazivnijih tumora sa večim rizikom za diseminaciju.

Actinic keratosis (AK) is proliferation of a neoplastic transformed keratinocytes confined to the epidermis and is considered as part of the initial clinical manifestation of the disease which can progress to squamous cell carcinoma (SCC), early stage in the biological spectrum SCC. The concept of progression with three possible steps in evolution from AK towards SCC is defined as a keratinocyte intraepidermal neoplasia (KIN), according to the level of epidermal involvement, analogous to cervical intraepithelial neoplasm (CIN). Squamous cell skin carcinoma in situ (SCCIS) or Bowen's disease (BD) is a malignant neoplasm confined to the epidermis, with full- thicknes keratinocyte atypia, but no signs of dermal invasion. Cutaneous SCC is the second most common malignant tumor of the skin, which is manifested as broad spectrum of lesions, from easily curable superficial to invasive and very infiltrated tumors that metastasize, leading to death. Although it can arise de novo, SCC, however, often arises from precursor lesions, AK and BD, which, as already mentioned, according to some authors actually represent SCC in earlier evolutionary stages. Although AK, SCCIS and SCC can be considered as different stages of the same disease not all the lesions progressed in SCC. Molecular mechanisms of progression have not yet been fully understood and explained. Minichromosome maintenance proteins (MCM) represent a complex of proteins that play a key role in the regulation of DNA replication during the cell cycle. MCM are group of the nuclear proteins involved in initiation and maintenance of DNA replication. Functional interactions between the MCM proteins and other components, by means of protein kinases which are involved in regulation of cell cycle, allows the DNA synthesis initiation, complete and accurate DNA replication to once per cell cycle, maintaining the genome stability. Expressed in cells in which the cell cycle is in progress, with lost of expression in quiescent and differentiated cells, MCM proteins represent potentially useful markers of cell proliferation, particularly in cancer. The aim of this study was to investigate MCM 2, 5 and 7 expression in AK, BD and invasive SCC, analysis of expression in situ lesions classified as keratinocyte intraepidermal neoplasia (KIN), comparison MCM 2, MCM5 and MCM7 expression and prognostic parameters of SCC such as size, depth of invasion, thickness and tumor grade, as well as the evaluation of different methods of evaluation and immunohistochemical staining in relation to the type of lesion and prognostic parametrs. The study was conducted on 315 cases, of which 91 were AK, 50 BD and 174 invasive SCC, of which an equal number of good (G1), medium (G2) and poorly (G3) differentiated SCC (58 cases). According KIN classification from 141 preinvasive lesions, 50 changes were KIN 1, 40 KIN 2 and 51 KIN 3. For MCM2, MCM5 and MCM7 expression immunohistochemiry (IHH) was done. Tissue microarray (TMA) was used for efficient, more economical, faster and easier analysis. The distributions of immunopositive cells were evaluated. Semiquantitative estimation of the number of positive cells was determined and expressed semiquantitatively. The MCM2, MCM5, and MCM7 immunostaining distribution was significantly different in AK, BD, and SCC . Basal expression of all three proteins was observed more frequently in AK, diffuse expression dominated in BD, and peripheral and diffuse distributions were observed in almost equal numbers in SCC. For in situ lesion classified as KIN, diffuse staining was more frequent in KIN3 than in KIN1, KIN2, or KIN1 and KIN2 combined and more frequent in KIN2 than in KIN1. All three proteins showed peripheral distribution in most SCC-WD cases and diffuse distribution in SCC-PD; peripheral and diffuse distributions were present in similar numbers of SCC-MD cases. Analysis of the immunostaining distribution in relation to SCC prognostic parameters showed more frequent diffuse staining in larger and thicker tumors, but the difference was statistically significant only for the 6-mm thickness threshold. Semiquantitative estimation of the number of positive cells showed the strongest association with lesion type using the 50% cut-off value. There was a statistically significant difference between AK, BD, and SCC, except for MCM7 between AK and SCC. There was an increase in the number of positive cells from KIN1 to KIN3 and from SCC-WD to SCC-PD. The most pronounced difference was between KIN 3 and KIN1, KIN2, and KIN1 and KIN2 combined, whereas the difference was less prominent when KIN1 and KIN2 were compared for MCM2 and MCM5, or did not reach statistical significance for MCM7. With respect to SCC grade, all MCM proteins showed highly significant differences between SCC-WD and both SCC-MD and SCC-PD and slightly less prominent differences between SCC-MD and SCC-PD. Analysis of the extent of immunostaining using the 50% threshold revealed statistically significant differences for all MCM proteins in relation to 6-mm thick SCC. No association between the size of the lesion and the level of MCM expression could be established. This is the first study in which distribution and expression of MCM proteins was evaluated in the keratinocyte skin neoplasms. Results from this analysis on MCM2, MCM5, and MCM7 expression in AK, BD, and SCC showed more prominent diffuse distribution and higher numbers of positive cells in BD than in AK, in KIN3 than in KIN1 and KIN2, and in SCC-PD than in SCC-WD, suggesting their role in the progression of both in situ and invasive cutaneous SCC. Comparison with SCC prognostic features revealed correlation only with 6 mm tumor thickness, which could imply association of MCM proteins with the high-risk factors in SCC. Finally, we observed similar results with all three MCM proteins, supporting them as suitable markers for the evaluation of progressing in situ lesions and possible indicators of invasive tumors with higher risk for metastatic dissemination.

Ispitivanje imunomodulatornog i citotoksičnog efekta vodenog ekstrakta bele imele (Viscum album L.) in vitro i u pacijentkinja na terapiji antraciklinskim hemioterapeuticima

Examination of immunomodulatory and cytotoxic effects of aqueous exstract of white mistletoe (Viscum album L.) in vitro and in patients treated with anthracycline chemotherapeutics

Uspešnost antitumorske terapije je ograničena pojavom tumorske rezistencije na lekove putem molekularnih mehanizama koji i dalje ostaju nerazjašnjeni. Kako bi se smanjili neželjeni efekti i poboljšala efikasnost standardnih hemioterapija i radioterapija, počeli su da se primenjuju mnogi komplementarni ili alternativni medikamenti. Bela imela (lat. Viscum album L) se tradicionalno koristi u lečenju pojedinih vrsta tumora. Terapijska korist od primene bele imele kada se koristi zajedno sa hemioterapijom ili radioterapijom, odnosno operacijom doprinosi ukupnom poboljšanju kvaliteta života kod pacijenata obolelih od tumora. Dosadašnja istraživanja na ćelijama u kulturi i na životinjskim modelima, su pokazala da potencijalni mehanizmi delovanja sastojaka bele imele uključuju antitumorsko, antiinflamatorno i imunomodulatorno dejstvo. Uprkos intezivnim ispitivanjima, mehanizam antitumorskog dejstva ekstrakta bele imele je još uvek u velikom obimu nepoznat. Takođe, malo je i naučnih podataka o mehanizmu interakcije bioaktivnih komponenti bele imele i drugih antitumorskih citotoksičnih lekova, koji bi pružili zasnovanost daljem kliničkom ispitivanju. Cilj ove disertacije je utvrđivanje imunomodulatornog dejstva ekstrakta bele imele na ćelije imunskog sistema u in vitro i in vivo uslovima, ispitivanje citotoksičnog efekta ekstrakta bele imele in vitro i molekularnih mehanizma interakcije bioaktivnih komponenti ekstrakta bele imele sa citotoksičnim agensom iz grupe antraciklina na modelu tumorskih ćelija u kulturi. Rezultati dobijeni u in vitro istraživanjima su pokazali citotoksični i antiproliferativni efekat ekstrakta bele imele, putem indukcije apoptoze tumorskih ćelija unutrašnjim mitohondrijskim putem i indukcijom zastoja u G0/G1 fazi ćelijskog ciklusa. Kombinovani tretman tumorskih ćelija subterapeutskom koncentracijom doksorubicina i ekstraktom bele imele dovodi do sinergističkog inhibitornog efekta. Apoptotska ćelijska smrti K562 i MCF 7 ćelija je istovremeno povezana sa smanjenim nakupljanjem ćelija u G2/M fazi ciklusa, koje se inače dešava pod delovanjem doksorubicina. Eliminacija G2/M zastoja, praćena povećanjem procenta ćelija u apoptozi, ima za posledicu aktivaciju kaspaze-3 kod K562 ćelija, prekomernu ekspresiju Bax, rani gubitak mitohondrijskog membranskog potencijala, prekomernu ekspresiju ciklina B1, sniženu ekspresiju ciklina D1, kao i sniženu gensku ekspresiju p21 i IGF-1R. Ekstrakt bele imele je pokazao i značajan inhibitorni efekat na markere aktivacije trombocita u in vitro uslovima, na agregaciju trombocita sa monocitima, kao i TNF-α posredovano antitumorsko dejstvo M1 makrofaga. Rezultati in vivo pilot studije nisu pokazali imunomodulatorno dejstvo ekstrakta bele imele na zastupljenost pojedinih subpopulacija limfocita u perifernoj krvi pacijentkinja obolelih od karcinoma dojke, ali je pokazano poboljšanje kvaliteta života datih pacijentkinja. Iako dizajn studije može da bude ograničenje, rezultati pokazuju izvodljivost studije koristeći opisane metode. U zaključku, naši rezultati ukazuju na to da ekstrakt bele imele dovodi do poboljšanja kvaliteta života pacijentkinja sa karcinomom dojke ako se primeni kao komplementarna terapija, kao i da ekstrakt bele imele može senzibilisati Bcr-Abl pozitivne K562 ćelije, kao i MCF7 ćelije na apoptotozu pri senescentnim, subapoptotskim koncentracijama doksorubicina. To nadalje znači da ukoliko se ekstrakt bele imele koristi kao komplementarna terapija, moguće je pri znatno nižim, manje toksičnim koncentracijama hemioterapeutika postići željeni efekat na tumorske ćelije, povećati antiproliferativnu aktivnost dokorubicina, suprimirajući terapijom indukovanu senesenciju.

Success of anticancer treatment has been impeded by resistance to anti-tumor therapy via molecular mechanisms that still remain elusive. To reduce side effects and improve the effectiveness of standard chemoradiation therapy, many complementary or alternative medicines (CAM) have been investigated. Mistletoe (Viscum album L) has been long used as CAM supporting cancer therapy. Therapeutic benefit of mistletoe preparations when utilized along with surgery, chemotherapy or radiotherapy contributes to the overall improvement in the quality of life in cancer patients. Several lines of evidence have revealed that mistletoe preparations in vitro and in vivo exert an anti-cancer, anti-inflamatory and immunomodulatory effects. Despite extensive experimental analyses of their biological properties, many questions regarding the precise mode of action of mistletoe still remain elusive. Also, there is only little information about interactions between mistletoe and anticancer drugs, which would provide validity for further clinical investigations. The aims of this study are investigation of immunomodulatory effects of mistletoe extracts on immune cells in vitro and in vivo, examination of cytotoxic effect in vitro, as well as molecular mechanisms underlying interactions between mistletoe and anticancer drug from the antraciklenes family on the tumor cells in culture. Our results from in vivo study study have shown cytotoxic and antiproliferative effect of mistletoe extract via induction of apoptosis of cancer cells through intrinsic mitochondrial pathway and induction of G0/G1 arrest. Also, combined treatment of cancer cells with sub-therapeutic concentration of doxorubicine and mistletoe extract led to synergistic inhibitory effect. Increased K562 and MCF7 cells apoptotic death during co- treatment was associated with reduced G2/M accumulation during doxorubicine treatment. The lack of G2/M arrest was accompanied by an increased percentage of apoptotic cells associated with cleavage of caspase-3 in the case of K562 cells, up-regulation of Bax expression level, early dissipation of mitochondrial membrane potential, up-regulation of cyclin B1, down-regulation of cyclin D1, as well as down-regulation of p21 and IGF-1R. Finally, in vitro mistletoe extract showed significant inhibitory effects on both platelet activation and aggregation with monocytes, as well as potential to activate M1 macrophage to produce TNF-α. In in vivo pilot study we reported no immunomodulatory effect of mistletoe extracts on immune cells of patients of breast cancer, but an improvement of quality of life of breast cancer patients was shown. Although the open design may be a limitation, the findings show the feasibility of a confirmatory study using the methods described here. In conclusion, our results suggest that mistletoe extract improves the quality of life of breast cancer patients if applied as CAM, as well as sensitizes Bcr - Abl positive K562 cells, and MCF7 cells on apoptotosis under senescent, subapoptotic concentrations of doxorubicin .This also means that if the white mistletoe extract is used as a complementary therapy, it is possible to by using much lower, less toxic concentrations of chemotherapeutics to achieve the desired effect on tumor cells via increment of the antiproliferative activity of doxorubicin, and suppressing therapy induced senescence.

Karakterizacija AggLb agregacionog faktora soja Lactobacillus paracasei subsp. paracasei BGNJ1-64

Characterization of AggLb aggregation factor from strain Lactobacillus paracasei subsp. paracasei BGNJ1-64

Odgovor bakterija na uticaj razliĉitih faktora sredine koja ih okruţuje, kao i njihova meĊusobna komunikacija, zavisi od procesa koji se odvijaju na površini bakterijskih ćelija. S obzirom na brojne mogućnosti primene bakterija roda Lactobacillus, kako u prehrambenoj industriji, tako i u medicini (u prevenciji i/ili leĉenju bolesti), potrebno je što bolje okarakterisati komponente koje ĉine površinski sloj ovih bakterija. Agregacija bakterija, determinisana faktorima lociranim na površini ćelija moţe se smatrati poţeljnom osobinom pri izboru probiotika, s obzirom da ima vaţnu ulogu u meĊubakterijskoj interakciji, u interakciji sa epitelijalnim ćelijama gastrointenstinalnog trakta i doprinosi sposobnosti bakterija da se adsorbuju na razliĉite površine, odnosno sposobnost agregacije omogućava uspešnu kolonizaciju staništa. Kolekcija laktobacila Laboratorije za molekularnu mikrobiologiju je pretraţena na prisustvo sojeva koji poseduju sposobnost agregacije (Agg+), nakon ĉega je za dalji rad odabrano jedanaest sojeva: BGSJ2-8, BGGR2-68, BGGR2-82, BGDP1-84, BGDP9-38, BGNJ1-3, BGNJ1-61, BGNJ1-64, BGNJ1-70, BGZLS30-6 i BGAR75. Odabrani Agg+ sojevi se na osnovu stepena agregacije i tipa formiranih agregata mogu podeliti u tri grupe: oni koji brzo, srednje i sporo agregiraju ili koji formiraju krupne, srednje i sitne agregate, pri ĉemu postoji direktna korelacija izmeĊu brzine agregacije i tipa agregata. Analizom sposobnosti vezivanja odabranih Agg+ sojeva laktobacila za komponente ekstracelularnog matriksa (ECM) zapaţen je širok dijapazon sposobnosti vezivanja za kolagen - od intenzivnog afiniteta vezivanja do odsustva sposobnosti vezivanja za matriks. Pritom su pojedini sojevi, BGGR2- 68 i BGGR2-82, ispoljili sposobnost vezivanja za plastiku (formiranje biofilma). Analizom plazmidnog sastava Agg+ sojeva konstatovano je prisustvo velikih plazmida u svim sojevima, a ĉišćenjem plazmida iz sojeva BGNJ1-64 i BGSJ2-8 potvrĊena je plazmidna lokacija gena odgovornih za ekspresiju agregacionog fenotipa. Nakon konstrukcije i analize plazmidnih biblioteka sojeva BGNJ1-64 i BGSJ2-8, izolovan je konstrukt pALb35 (koji nosi SacI fragment veliĉine 11,4 kb poreklom iz plazmida pNJ1 soja BGNJ1-64) koji dovodi do ponovne pojave autoagregacionog fenotipa transformisanog heterologog domaćina BGKP1-20 (Agg-). Sposobnost autoagregacije derivata BGKP1-20/pALb35 je posledica ekspresije aggLb gena koji se nalazi u okviru SacI fragmenta, konstrukta pALb35 korišćenog za transformaciju soja BGKP1-20. AggLb protein je najveći agregacioni faktor laktobacila do sada opisan u literaturi, veliĉine 318,6 kDa. Pored domena neophodnih za transport kroz membranu i ukotvljavanje, AggLb protein sadrţi veći broj ponovaka dva razliĉita tipa domena koji su veoma znaĉajni za sposobnost adhezije: kolagen vezujući domeni na N- terminusu proteina (6 domena koji meĊusobno pokazuju veliku heterogenost; sa stepenom homologije manjim od 26%) i CnaB-like domeni (kolagen vezujući domeni B) na C- terminusu proteina (20 puta ponovljeni, skoro 100% identiĉni, aminokiselinski ponovci). U cilju razumevanja uloge, funkcije i stepena uticaja agregacionog faktora na probiotiĉki potencijal soja BGNJ1-64 uraĊeni su eksperimenti funkcionalne karakterizacije AggLb proteina. Pokazano je da AggLb protein eksprimiran na površini heterologog domaćina BGKP1-20, ima ulogu u autoagregaciji i interakciji sa komponentama ECM-a, kao i u poslediĉnoj inhibiciji vezivanja patogenih sojeva za kolagen, dok njegov doprinos u koagregaciji patogenih sojeva Staphylococcus aureus, Escherichia coli O157:H7 i Listeria innocua ATCC33090 nije detektovan. U in vitro eksperimentu je pokazano da prisustvo AggLb proteina na površini ćelija doprinosi u velikom stepenu ekskluziji patogena: S. aureus (62,87%) i E. coli O157:H7 (57,01%). Konstrukcijom varijanti AggLb proteina koji u razliĉitim kombinacijama sadrţe razliĉit broj kolagen vezujućih domena (6, 2 (hibridi I-V i VI domen), 1+1/2 (1/2 V i VI domen) ili 0 (bez domena)) i CnaB-like domena (20, 10, 7, 5 ili 2), omogućeno je sagledavanje doprinosa pojedinaĉnih domena u okviru AggLb proteina u ostvarivanju njegovih pozitivnih probiotiĉkih efekata. Konstrukcijom hibridnog proteina koji sadrţi prvi deo kao kod laktokokalnog agregacionog faktora i drugi deo kao laktobacilusni faktor agregacije, omogućeno je praćenje doprinosa razliĉitih tipova kolagen vezujućih motiva u interakciji sa receptornim molekulima. Pokazano je da od analiziranih derivata jedino varijanta AggLb proteina, eksprimirana na površini bakterijske ćelije koja sadrţi kolagen vezujuće domene II, III i IV, je bila sposobna da indukuje autoagregaciju, vezivanje za kolagen i fibronektin ukazujući da su ova tri domena najodgovornija za navedene funkcije. CnaB-like domeni najverovatnije nemaju direktnu ulogu u agregaciji i vezivanju za kolagen i fibronektin, već se njihov doprinos ogleda u pozicioniranju kolagen vezujućih domena što dalje od površine ćelije kako bi se što efikasnije omogućila interakcija ovih domena sa receptornim molekulima koji mogu biti na razliĉitoj udaljenosti od bakterijskih ćelija unutar ECM-a. TakoĊe, pokazano je da je tip kolagen vezujućih domena prisutnih u agregacionom proteinu vaţniji od broja njihovih ponovaka. Primećeno je da uklanjanje odreĊenih domena u AggLb proteinu ne samo da onemogućava njegove funkcije autoagregacije i vezivanja za komponente ECM-a kod pojedinih derivata koji na površini eksprimiraju odreĊene varijante AggLb proteina, već istovremeno rezultira mogućnošću pojedinih domena AggLb proteina da doĊu do izraţaja (bivaju demaskirani otklanjanjem drugih domena) i preuzmu funkciju formiranja biofilma. Delecija pojedinih domena prevodi AggLb protein iz jednog kompleksa funkcija koje su direktno povezane sa prisustvom odreĊenjih domena u drugu funkciju koja nije moţda tako intenzivna kao prethodne ali je zasigurno znaĉajna ukazujući da proteini mogu imati širi potencijal funkcija nego što mi na prvi pogled vidimo.

Response of bacteria to different environmental factors as well as their communication primarily depends on the processes that occur on their cell surface. Due to the continuously increasing number of application possibilities of lactobacilli, both in the food industry and in medicine (in the prevention and/or treatment of the disease), there is a need for a more comprehensive characterization of the components that form the surface layer of these bacteria. Aggregation ability of bacteria, determined by factors located on the cell surface could be considered as a desirable trait in the selection of probiotics. Since it plays an important role in the interaction between bacteria, in interaction with the epithelial cells of the gastrointestinal tract plus contributing to the bacterial capability to adhere to different surfaces, which is an important feature enabling them to successfully colonize diverse habitats. Lactobacilli collection of Laboratory for molecular microbiology was screened for the presence of strains possessing the aggregation ability (Agg+) and for the further work eleven strains have been selected: BGSJ2-8, BGGR2-68, BGGR2-82, BGDP1-84, BGDP9-38, BGNJ1-3, BGNJ1-61, BGNJ1-64, BGNJ1-70, BGZLS30-6 and BGAR75 which, according to the level of aggregation and the appearance of the aggregates formed, could be clasified into three groups: the fast, the medium and the slow aggregating ones or into the groups of those forming large, medium or small aggregates wherein a direct correlation between the aggregation kinetics and the type of the formed aggregates was found. By analyzing the binding ability of the selected Agg+ strains of lactobacilli to the components of extracellular matrix (ECM) a wide range of binding capacity to collagen was observed, varying from the intensive to the complete lack of the binding capacity to the matrix. Two strains (BGGR2-68 and BGGR2-82) showed the ability to bind even to plastics (form biofilms). By analyzing the plasmid composition of Agg+ strains, the presence of large plasmids was detected in all strains. The plasmid curing experiments confirmed the plasmid location of the gene(s) responsible for the expression of aggregation phenotype in strains BGNJ1-64 and BGSJ2-8. After the construction and analysis of BGNJ1-64 and BGSJ2-8 strains plasmid libraries, selected construct pALb35 (which carries the SacI fragment of 11.4 kb. derived from plasmid pNJ1 from strain BGNJ1-64) was able to reconstitute the autoagreggation ability of the transformed heterologous host BGKP1-20 (Agg-). The autoaggregation ability of the derivative BGKP1-20/pALb35 was the result of expression of aggLb gene located within the SacI fragment in the construct pALb35 used for transformation of BGKP1-20 strain. AggLb protein is the largest aggregation factor in lactobacilli so far described in the literature, with the molecular mass of 318.6 kDa. In addition to all sequences necessary for the export and anchoring, AggLb protein contains a number of repeats of two different types of domains, which are important for the adhesion, namely the collagen-binding domains at the N-terminus of the protein (6 domains, with great heterogeneity, with less than 26% of homology) and the CnaB-like domains (collagen binding domain B) at the C-terminus of the protein (repeated 20 times, with an almost 100% identical amino acid repeats). In order to understand the aggregation factor role, function and level of contribution to the probiotic potential of strain BGNJ1-64, experiments of functional analysis were done. It was shown that AggLb protein expressed on the surface of the heterologous host BGKP1- 20, plays a role in autoagreggation and interaction with the components of the ECM, consequently inhibiting binding of pathogenic strains to the collagen, while its impact on the coaggregation with pathogenic strains Staphylococcus aureus, Escherichia coli O157:H7 and Listeria innocua ATCC33090 was not detected. In vitro experiments demonstrated that the presence of the AggLb protein on the cell surface contributes greatly to exclusion of pathogens: S. aureus (62,87%) and E. coli O157:H7 (57,01%). Constructed variants of AggLb protein designed to have different combinations with different numbers of repeats of the collagen binding domains (6, 2 (hybrids domains IV and VI), 1 + 1/2 (2.1 V and VI domains) or 0 (no domains)) and CnaB- like domains (20, 10, 7, 5 and 2) enabled the assessment of the contribution of the individual domains within the AggLb protein in achieving its probiotic effect. By construction of a hybrid protein containing the proximal part of the lactococal aggregation factor (AggL) and the distal part consisted of lactobacilli aggregation factor (AggLb) it was possible to monitor the contribution of the different types of collagen binding motifs in the interaction with the receptor molecules. It is shown that among the analyzed derivatives only a variant of the AggLb protein expressed on the surface of bacterial cells containing the collagen binding domains II, III and IV was able to induce autoaggregation, binding to collagen and fibronectin, indicating that these three domains are those the most responsible for the functions. CnaB-like domains likely has no direct role in the aggregation and binding to collagen and fibronectin, but their contribution is reflected in pulling forward collagen binding domains away from the cell surface in order to allow more efficient interaction of the domains with the receptor molecules so that they could be positioned at different distances from the bacterial cells within EMC. Also, it was shown that the type of collagen binding domains present in the aggregation protein is more important than the number of their copies. It was noted that the removal of certain domains in the AggLb protein not only prevents its autoagreggation and binding to components of ECM in certain derivatives expressing specific variants of AggLb protein on the surface, but also results in the ability of some protein domains of the AggLb to come to the fore (are unmasked by elimination of other domains) and assume the function of biofilm formation. Deletion of certain domains switches the AggLb protein from one set of functions (that are directly related to the presence certain domains) to an another function that is not as intense as the previous one but is certainly significant, indicating that the proteins can have a wider range of functions than we see at the first glance.

Vanćelijski metabolizam adeninskih nukleotida u sinaptozomima hipokampusa pacova - polne specifičnosti i uloga ženskih polnih hormona

Metabolism of extracellular adenine nucleotides in rat hippocampal synaptosomes - sex differences and the roll of female sex steroids

Adenozin-5-trifosfat (ATP) u centralnom nervnom sistemu (CNS) ima ulogu brzog ekscitatornog neurotransmitera i kotransmitera ali i trofičkog faktora, gliotransmitera i signalnog molekula koji učestvuje u komunikaciji između ćelija CNS. Kada se oslobodi u vanćelijski prostor, npr. sinaptičku pukotinu i aktivira odgovarajuće receptore (P1 i P2), ATP se brzo degradira, posredstvom enzima ektonukleotidaza. Primarna uloga ovih enzima je sekvencijalna hidroliza adeninskih nukleotida, kao što su ATP, adenozin-5-difosfat (ADP) i adenozin-5-monofosfat (AMP) čime nastaje nukleozid adenozin, potentni neuromodulator i homeostatski regulator u CNS. S obzirom na to da koncentracije vanćelijskog ATP i adenozina direktno zavise od stepena aktivnosti ektonukleotidaza, svaka promena ekspresije i aktivnosti ovih enzima može biti relevantna u kontekstu fiziologije i patologije. Oskudni literaturni podaci ukazuju da su ektonukleotidaze deo složene molekulske mreže koja je pod uticajem polnih hormona. Kako ženski polni hormoni, pogotovo 17β-estradiol (E2), utiču na gustinu sinapsi, složenost dendritskih grananja i broja trnolikih izraštaja u hipokampusu, ostvarajući tako snažan uticaj na sinaptičku plastičnost, učenje i pamćenje kod pacova oba pola, u ovoj tezi ispitan je uticaj hormonskog statusa i mehanizam regulacije/modulacije sinaptičkih ektonukleotidaza, ekto- nukleozid 5'-trifosfat difosfohidrolaze 1-3 (NTPD-aza 1-3) i ekto-5'-nukleotidaze (eN) 17β- estradiolom u hipokampusu ženki i mužjaka pacova. Takođe, ispitan je uticaj ponovljenog (sedmodnevnog) tretmana 17-estradiolom (E), E2 i progesteronom (P4) na aktivnost i ekspresiju eN u totalnoj membranskoj frakciji hipokampusa oba pola. Rezultati biohemijskih analiza ukazuju da aktivnost ispitanih ektonukleotidaza diskretno fluktuira tokom estrusnog ciklusa, kao i da uklanjanje jajnika (OVX), primarnog izvora ženskih polnih hormona, smanjuje nivo hidrolize ATP, ADP, AMP u sinaptozomima hipokampusa ženki pacova. Ovi nalazi jasno ukazuju na to da su ektonukleotidaze (NTPD- aza 1-3 i eN) pod regulatornom kontrolom endogenih hormona jajnika. Promena stepena hidrolize adeninskih nukleotida uočena je kod OVX ženki nakon jednokratnog tretmana E2, pri čemu je stepen hidrolize ATP i ADP povećan verovatno kao rezultat povećane ekspresije NTPD-aze 1 i 2, dok relativna zastupljenost NTPD-aze 3 ostaje nepromenjena. E2 takođe dovodi do porasta nivoa hidrolize AMP pri čemu reguliše/moduliše eN aktivacijom klasičnih unutarćelijskih estradiolskih receptora (ER i ERβ) koji različitim mehanizmima dovode do povećanja aktivnosti eN u sinaptozomima hipokampusa OVX ženki. E2 moduliše vanćelijski metabolizam adeninskih nukleotida i u sinaptozomima hipokampusa mužjaka pacova. E2 smanjuje proteinsku ekspresiju i/ili aktivnost NTPD-aza 1, 2 i eN, ali ne menja relativnu zastupljenost NTPD-aze 3 u sinaptozomima hipokampusa mužjaka. Uprkos smanjenju aktivnosti NTPD-aza, E2 ne izaziva štetnu akumulaciju ATP i ADP, jer povećana aktivnost TNAP, nakon tretmana E2, ukazuje na potencijalnu eliminaciju viška ATP iz sinaptičke pukotine koji nastaje smanjenjem aktivnosti NTPD-aza 1-2, odnosno višak ADP koji se akumulira usled nepromenjene aktivnosti NTPD-aze 3. E2 na nivou eN ostvaruje kompetitivni tip inhibicije što ukazuje na moguću interakciju E2/eN u membrani. Takođe, eN je modulisana aktivacijom unutarćelijskih ER i ERβ, ali i pokretanjem mehanizama nezavisnih od klasičnih ER. Skoro 2,5 puta veća aktivnost eN izmerena je u P2 frakciji izolovanoj iz hipokampusa mužjaka pacova u odnosu na ženke u disetrusnoj fazi ciklusa i OVX. Rezultati analiza ekspresije totalne iRNK, kao i proteina i aktivnosti eN u membranskoj frakciji ukazuju da sedmodnevni tretmani E2 i P4 ispoljavaju različite efekte na nivou genske ekspresije eN u hipokampusu OVX ženki pacova, dok iste efekte ispoljavaju na nivou relativne količine proteina i aktivnosti eN. Za razliku od ženki, u hipokampusu intaktnih mužjaka, nakon hroničnih tretmana steroidnim hormonima, nije zabeležena promena eN ni na jednom ispitivanom nivou. Može se zaključiti da su ektonukleotidaze u sinapsama hipokampusa oba pola regulisane/modulisane E2 ili samim procesima koje E2 inicira. Rezultati ove doktorske teze govore u prilog tome da poznati efekti E2 na nivou funkcionalnih karakteristika sinapsi u hipokampusu, mogu delom biti posledica zapaženih efekata na nivou ektonukleotidaza, enzima odgovornih za vanćelijsko formiranje adenozina u CNS.

Adenosine-5´-triphosphate (ATP) is an important extracellular signaling molecule. It acts as neurotransmitter, co-transmitter, gliotransmitter and trophic factor in the central nervous system (CNS). Upon the release, ATP modulates synaptic transmission by acting at either ionotropic (P2X) or metabotropic (P2Y) receptors, and is sequentially catabolized by the action of ectonucleotidases. The first step of ATP inactivation is mediated by the family of ecto-nucleoside triphosphate diphosphohydrolase (NTPDase), which are able to hydrolyze ATP and adenosine-5´-diphosphate (ADP) to adenosine-5´-monophosphate (AMP). The last and the rate-limiting step in of the ATP conversion is catalyzed by ecto-5´- nucleotidase (eN), which hydrolases AMP to adenosine, potent neuromodulator and homeostatic regulator in the CNS. Since ectonucleotidases is crucial for maintaining of physiological levels of extracellular adenine nucleotides in the CNS, any alteration in their activity and expression can be relevant in context of physiology and pathology. Literature data indicate that ectonucleotidases may be part of complex molecular network which may be modulated by ovarian steroids. Based on this findings, and the fact that female sex steroids, particularly 17β-estradiol (E2) play an essential role in the modulation of hippocampal synaptic plasticity therefore influencing hippocampal dependent learning and memory, we hypothesized that steroid hormones, in particular E2 has potential to modulate the activity and expression of ectonucleotidases (NTPDase 1-3 and eN) in hippocampal synaptosomes of male and female rats. Also, the aim of this doctoral dissertation was to determine the impact of steroid hormones on eN after repeated administration of 17α- estradiol, E2 and progesterone (P4) for seven consecutive days in hippocampus of both sexes. It is shown that the activity of the examined ectonucleotidases fluctuates across the estrous cycle in the hippocampal synaptosomes of female rats, while significant reduction level of ATP, ADP and AMP hydrolysis is observed in hippocampal synaptosomes after bilateral ovariectomy. These results clearly indicate that examined ectonucleotidases are regulated/modulated by endogenous female sex steroids. Biochemical analysis indicate that acute E2 treatment in the OVX rats significantly increase the level of ATP and ADP hydrolysis, probably as a result of upregulated NTPDase 1 and 2, while E2 had no effect on NTPDase 3. The level of AMP hydrolysis was also augmented in hippocampal synaptosomes of OVX rats after E2 treatment. Our initial evaluation imply that regulatory E2 action at the activity and protein abundance of eN is mediated by both intracellular estradiol receptors (ERα and ERβ) through different mechanisms in hippocampal synaptosomes of OVX rats. E2 also influence extracellular metabolism of adenine nucleotides in hippocampal synaptosomes of male rats. E2 downregulates expression and/or activity of NTPDase 1, NTPDase 2 and eN, and does not affect the NTPDase 3. Despite the reduction of NTPDase activity, E2 did not induce toxic accumulation of ATP and ADP in the extracellular space, since E2 treatment resulted in increased TNAP activity in male hippocampal synaptosomes, which indicates effective elimination of ATP and ADP in the synaptic cleft. In male hippocampal synaptosomes, E2 induce competitive inhibition of eN, which indicates possible E2/eN interaction in the cell membrane. eN activity might be modulated through activation of classic intracellular estradiol receptors and also through other mechanism which are independent of classical ER. In the hippocampus obtained from male rats, more than 2.5-fold higher AMP hydrolysis was observed compared to females in diestrus state and OVX. Results obtained on eN-mRNA, protein abundance and eN activity level indicate that repeated E2 and P4 induced different effects on eN gene expression in hippocampus of OVX rats, while similar effects were observed on the level of protein expression and activity. In male rats, administered sex steroids did not affect hippocampal eN at any analyzed level. In general, we can conclude that E2 differentially modulate/regulate NTPDase 1- 3 and eN in hippocampal synaptosomes of both sexes. These results may imply that known sex-specific effects of E2 on functional properties of hippocampal synapses may be in part mediated through observed action on ectonucleotidases, which participate in the formation of extracellular adenosine.

Biohemijski i ekofiziološki odgovori odabranih vrsta mahovina na kontrolisani stres izazvan solima u akseničnim uslovima

Biochemical and ecophysiological properties of selected moss species during salt stress in the axenical conditions

Stres soli je, pored suše, jedan od najznačajnijih abiotičkih faktora koji ograničava produktivnost biljaka, a utiče negativno i na čitave ekosisteme...

Salt stress is, among drought, one of the most important abiotic factors that limits plant productivity and has negative effect on the ecosystems...

Transformacija industrijskih prostora u kontekstu post-socijalističke tranzicije grada Skoplja

Transformation of industrial space in the context of post-socialist transition of the city of Skopje

Teza se bavi istraživanjem transformacije industrijskog urbanog prostora u Skoplju u post-socijalističkom periodu. U užem smislu, rad se bavi istraživanjem transformacije industrijskog prostora, odnosno intenzivnim restruktuisanjem prostora proizvodnih aktivnosti usled delovanja procesa deindustrijalizacije i reindustrijalizacije, pojavom novih prostornih formi proizvodnih aktivnosti i novim prostornim razmeštajem industrije. Istraživanje polazi od stava da je transformacija industrijskog prostora u Skoplju kao proces sličan onim kod gradova u Centralnoj i Istočnoj Evropi, uslovljenim procesima ekonomskog restruktuisanja, deindustrijalizacije, globalizacije i tercijarizacije ekonomije. Ipak, specifike koje proizlaze iz tranzicije, nivo ekonomske razvijenosti i mesto Skoplja u regionalnim i globalnim ekonomskim tokovima određuju precizne i jasne razlike kod procesa transformacije. Polazne hipoteze u ovom istražuvačkom radu su da se transformacija socijalističkog industrijskog prostora karakteriše kombinacijom kontinuiteta i promena, i da je lokacija nove industrijske aktivnosti rezultat delovanja procesa koncentracije i disperzije proizvodnih aktivnosti. Kroz istraživanje kontinuiteta i diskontinuiteta u razvoju i razmeštaju industrije i industrijskih područja u gradu Skoplju, tendencije koncentracije i disperzije industrijskih aktivnosti, pojava novih prostornih formi proizvodne i privredne aktivnosti, i promena vi prostornih obrazaca industrije potvrdiće se da su dominatni ishod transformacije industrijskog prostora lokacije napuštene industrije (promena) i da disperzni razmeštaj industrijske aktivnosti van urbanih granica grada ima veću ulogu u pogledu prostornog usmeravanja novog industrijskog razmeštaja. Doprinos istraživanja je dvojak. Prva grupa rezultata istraživanja potvrđuje se, na primeru grada Skoplja, povezanost između transformacije prostora i razvoja društva u okviru šireg teorijskiog diskursa restruktuisanja prostora proizvodnje kod socijalističkog grada sa tranzicijom ekonomije i promenom političkog i društvenog sistema. Druga grupa rezultata istraživanja upućuje na potencijal i značaj postojećih i novostvorenih industrijskih područja za budući prostorni i privredni razvoj grada. Kroz teorijsko i empirijsko znanje o lokalnim problemima mogu se ponuditi praktični odgovori u oblasti arhitektonskog projektovanja i urbanističkog planiranja jer planiranje i projektovanje industrije treba da se bazira na poimanju realnih urbanih procesa usled delovanja faktora: istorijski, ekonomski i geopolitički kontekst, kao i društveni i ekološki problemi. Odgovornost budućeg razvoja grada sa urbano-arhitektonskog aspekta je u pronalaženju mogućnosti za obnovu i izgradnju unutar postojećeg urbanog konteksta i kompleksnog ekološkog okruženja kako bi se postigao željeni održiv razvoj grada.

Thе thesis explores the transformation of industrial urban space in Skopje in the post-socialist period. More specifically, the paper studies the industrial space transformation, i.e. the intensive spatial restructuring of production activities due to the effect of the processes of deindustrialization and reindustrialization, the emergence of new special forms of production activities and the new spatial distribution of industry. The research builds upon the statement that as a process, the transformation of industrial space in Skopje is similar to the one in cities of Central and Eastern Europe, where it resulted from the processes of economic restructuring, deindustrialization, globalization and tertiarization of the economy. Nevertheless, specifics that derive from the transition process, the low level of economic development and the position of Skopje within the global economic flows create precise and clear distinctions in the transformation process. The basic research hypotheses refer to the transformation of socialistic industrial space as characterized by a combination of continuity and change, and the location of new industrial activity resulting from the effects of processes of concentration and dispersion of production activities. Through the research of continuity and discontinuity in the development and distribution of industry and industrial areas in the city of Skopje, the tendencies of concentration and dispersion of industrial activities, the emergence of new spatial forms of production and economic activities, and the change of the spatial pattern of distribution of industry indicates that the predominant outcomes of transformation of the industrial space are viii locations of abandoned industries (change) and that the dispersed distribution of industrial activities beyond the urban boundaries of the city play a greater role in directing the new distribution of industry. The contribution of the research is twofold. The first set of research results confirm, in the case of the city of Skopje, the correlation between the transformation of the space and the development of society within the framework of general theoretical discourse of restructuring the production space in the socialist city, with transition of the economy and changes in the political and social system. The second set of research results point to the potential and importance of existing and newly established industrial areas for future spatial and economic development of the city. Drawing on the theoretical and empirical knowledge of local issues, practical responses in the field of architectural design and urban planning can be offered. This is based on the premise that industrial planning and design should be based on a comprehensive real urban process, which will take into account the effect of the historical,economic and geopolitican context. As well as social and ecological issues. From an urban-architectural perspective, it should be paramount the future development of the city to offer opportunities for renewal and construction within the existing urban context and complex ecological environment in order to achieve the desired sustainable development of the city.

Selekcija linija kukuruza sa poboljšanim kvalitetom proteina i adaptiranih na umereno klimatsko područje upotrebom molekularnih markera

Marker assisted selection for quality protein maize inbred lines adapted to temperate regions

Hranljiva vrednost kukuruza je niska zbog niskog sadržaja dve esencijalne aminokiseline - lizina i triptofana. Poboljšanje hranljive vrednosti kukuruza predstavlja jedan od glavnih ciljeva savremenih programa oplemenjivanja zbog sve značajnijih klimatskih promena i rasta ljudske populacije, odnosno sve veće potrebe za hranom. Najveći uspeh postignut je 60-ih godina prošlog veka otkrićem opaque2 (o2) mutanta sa superiornim hranljivim svojstvima. Međutim, plejotropni efekti o2 mutacije čine endosperm zrna kukuruza mekim, a samo zrno osetljivim na pucanje, trulež i štetočine. Istraživači Međunarodnog centra za poboljšanje pšenice i kukuruza u Meksiku (eng. International Maize and Wheat Improvement Center - CIMMYT) su metodama konvencionalnog oplemenjivanja stvorili kukuruz visokog kvaliteta proteina (eng. Quality Protein Maize – QPM), koji istovremeno ima visoku hranljivu vrednost proteina endosperma i dobre agronomske performanse. Iako je QPM razvijen klasičnim metodama selekcije, poslednjih godina se sve više primenjuje selekcija upotrebom molekularnih markera (eng. Marker Assisted Selection - MAS) sa ciljem poboljšanja elitnih linija za kvalitet proteina. Unutar sekvence o2 gena identifikovana su tri različita SSR markera – phi057, phi112 i umc1066, koji se koriste za utvrđivanje polimorfizma inbred linija i kao selekcioni markeri za o2 gen (eng. foreground selection). Molekularni markeri se koriste i radi utvrđivanja procenta genoma rekurentnog roditelja u potomstvima povratnih ukrštanja (eng. background selection). Primena foreground i background selekcije smanjuje broj generacija potrebnih za stvaranje željenog genotipa klasičnim metodama selekcije koje se zasnivaju na fenotipskom odabiru biljaka. Osnovni cilj ovog rada bio je dobijanje linija kukuruza poboljšanog kvaliteta proteina adaptiranih na umereno klimatsko područje, korišćenjem specifičnih molekularnih markera za opaque2 gen, kao i da se održe dobre agronomske osobine i kombinaciona sposobnost poboljšanih linija. Za poboljšanje kvaliteta proteina korišćene su, kao rekurentni roditelji (recipijenti o2 recesivne mutacije), elitne linije standardnog kvaliteta zrna ZPL 3 i ZPL 5. Kao izvor povećanog sadržaja triptofana i lizina (donor o2 recesivne mutacije) korišćena je tropska QPM linija CML 144. Identifikacija i potvrda rezultata QPM selekcije podrazumevala je biohemijske analize (određivanje sadržaja proteina, triptofana i izračunavanje indeksa kvaliteta - procenta triptofana u proteinu), kao i evaluaciju agronomski važnih osobina u poljskim ogledima. Kao rezultat ovog rada dobijeno je trinaest linija povećanog sadržaja triptofana (od 37-50% u odnosu na početnu vrednost roditeljske standardne linije), povećanog indeksa kvaliteta (42-56% u odnosu na početnu vrednost roditeljske linije), očuvane genetičke sličnosti sa rekurentnom linijom (93% genoma rekurentnog roditelja), visokog stepena modifikacije endosperma zrna (100% zrna tvrdog endosperma), povećanog prinosa zrna (11-31% u odnosu na roditeljsku liniju) i očuvanih odličnih kombinacionih sposobnosti. Biohemijski parametri su pokazali pozitivne korelacije sa prinosom zrna, što ukazuje na mogućnost paralelne selekcije na povećani prinos i poboljšanje kvaliteta zrna u ispitivanom materijalu. Na osnovu dobijenih korelacija između ispitivanih biohemijskih i fenotipskih osobina, za dobijanje genotipova sa većim sadržajem triptofana i indeksa kvaliteta mogla bi se preporučiti selekcija na visinu biljke, dužinu klipa i broj redova zrna. Između hibrida sa roditeljskom linijom ZPL 5 i hibrida sa odabranim BC2F3 potomstvima, za većinu ispitivanih fenotipskih osobina nisu utvrđene statistički značajne razlike, što je ukazalo da su kod sub-linija poboljšanog kvaliteta očuvane dobre kombinacione sposobnosti roditeljske linije ZPL 5. Mali procenat zrna sa mekim endospermom kod ispitivanih hibrida potvrdio je prisustvo gena modifikatora za tvrdoću endosperma. Kombinacijom konvencionalnog i molekularnog oplemenjivanja dobijene su linije poboljšanog kvaliteta proteina, koje su adaptirane na uslove umerenog klimatskog područja i mogu se preporučiti za programe stvaranja QPM hibrida.

Maize nutritional quality is poor due to the low levels of two essential aminoacids - lysine and tryptophan. Because of the climatic changes and increased food demands due to the human population growth, improvement of the maize nutritional quality is one of the main goals of many breeding programs. A major breakthrough has been the isolation of the opaque2 (o2) mutant with superior nutritional properties in the 1960s. However, pleiotropic effect of the o2 mutation makes the maize endosperm soft and susceptible to cracking, ear rots, and storage pests. Using conventional breeding methodologies, interdisciplinary research team in the International Maize and Wheat Improvement Center (CIMMYT), Mexico, created the new, agronomically acceptable and nutritionally improved opaque2 types by the name of Quality Protein Maize (QPM). Although QPM was created through conventional breeding, marker assisted selection (MAS) is becoming increasingly used approach for improvement of protein quality in maize. Three simple sequence repeats (phi057, phi112 and umc1066), located as internal repetitive sequences within the o2 gene, are being utilized as foreground selection markers for the o2 gene. Molecular markers are also effectively employed for identification of the genotypes with the highest proportion of recurrent parent genome (background selection). Both foreground and background selection decrease the number of the generations required to create desirable genotype through conventional breeding based on the fenotypic selection. The main objective of this research was to create high quality protein maize lines adapted to temperate regions using opaque2 specific molecular markers, while maintaining their good agronomic performances and combining abilities. Two elite inbred lines - ZPL 3 and ZPL 5 were selected as the recurrent parents, to be the recipients of the o2 allele and to improve the tryptophan content. CML 144, a QPM tropical inbred line, was selected as the donor line of o2. Identification and confirmation of the QPM selection results involved biochemical analysis (determination of protein content, tryptophan content and quality index - tryptophan to protein ratio) and evaluation of the agronomicaly important traits in the field trials. As the result of this research, 13 lines were created that have 37-50% increased tryptophan content compared to their normal version, 42-56% increased quality index, 93% recovery of the recurrent parent genome, 100% kernels with hard endosperm, 11- 31% increase in grain yield and preserved combining abilities. Positive correlations were found between grain yield and biochemical parameters, indicating potential simultaneous selection and improvement for these traits. Based on the correlations found between biochemical and phenotypic parameters, selection for the plant height, ear length and kernel row number could be recomended for development of high quality maize genotypes. Significant differences were not found for the majority of the agronomic traits between hybrids created with ZPL 5 and hybrids with selected BC2F3 progenies, what has confirmed that combining abilities of the parental line ZPL 5 are preserved. Also, small percentage of the soft endosperm kernels of the analyzed hybrids confirmed the presence of the modifier genes that cause the formation of a hard, vitreous endosperm. Using the integrated conventional and molecular breeding approach, we created high protein quality maize lines adapted to temperate regions, that can be considered as candidate parents for developing QPM hybrids.

Procena ekološkog statusa reke Zasavice na osnovu algoloških parametara

Ecological status assessment of the Zasavica river based on algological parameters

Jedinstven vodeni ekosistem Zasavica, koji čini kičmu istoimenog Specijalnog rezervata prirode, u svojoj osnovi podrazumeva rečni ekosistem, a danas predstavlja napušteno rečno korito, odnosno vlažno stanište, uporedivo u ekološkom smislu sa plitkim jezerima, barskim i močvarnim ekosistemima. Postizanje i održavanje dobrog ekološkog statusa vlažnih staništa, gde je ekološki status odraz kvaliteta strukture i funcionalnosti tih ekosistema, predstavlja glavni cilj Direktive o vodama. U Srbiji procenu ekološkog statusa površinskih voda reguliše Pravilnik o parametrima ekološkog i hemijskog statusa površinskih voda i parametrima hemijskog i kvantitativnog statusa podzemnih voda. Predmet ove doktorske disertacije je detaljna analiza cijanobakterija i algi sa ciljem procene ekološkog statusa Zasavice na osnovu parametara fitoplanktona i fitobentosa kao relevantnih bioloških grupa. Dodatno je analizirana i zajednica metafitona, da bi se dobili dopunski podaci o stabilnosti ovog osetljivog ekosistema. Uzorci za analizu fizičkih, hemijskih i biohemijskih (koncentracija hlorofila a) parametara kvaliteta vode, kao i uzorci za kvalitativnu i kvantitativnu analizu fitoplanktona, fitobentosa (epilitskih silikatnih algi) i metafitona, prikupljani su jednom mesečno od decembra 2012. godine do novembra 2013. godine, na dva lokaliteta. Nakon prikupljanja uzoraka, usledila je laboratorijska analiza uzoraka standardnim metodama i izračunavanje potrebnih indeksa. Kvalitativnom analizom cijanobakterija i algi utvrđeno je prisustvo 503 taksona iz 8 razdela (Cyanobacteria, Bacillariophyta, Chlorophyta, Chrysophyta, Cryptophyta, Dinophyta, Euglenophyta i Xanthophyta). Razdeo Bacillariophyta se izdvaja sa 170 identifikovanih taksona. Od 50 prisutnih taksona cijanobakterija, 12 je novo za floru Srbije. Pored toga, 13 taksona cijanobakterija je potencijalno toksično, a među njima tri vrste su i invazivne na području Evrope (Cylindrospermopsis raciborskii, Raphidiopsis mediteranea i Sphaerospermopsis aphanizomenoides). Od ukupno 89 zabeleženih taksona iz razdela Euglenophyta, njih 9 predstavlja nove taksone za floru Srbije. Imajući u vidu da je 28 identifikovanih taksona razdela Chrysophyta više od polovine do sada poznatih taksona zlatnih algi Srbije, Zasavica se može okarakterisati kao stanište sa izuzetnim bogatstvom ove grupe algi, a čak 16 taksona zabeleženo je po prvi put u Srbiji. Pored velikog specijskog, primećen je i veliki funkcionalni diverzitet fitoplanktona, koji je sagledan kroz tri različita klasifikaciona sistema. Upotrebom funkcionalnih klasifikacija fitoplanktona dobijaju se podaci uporedivi na globalnom nivou i olakšava se interpretacija rezultata ekoloških istraživanja akvatičnih ekosistema. Dovoljna količina nutrijenata, same geomorfološke i hidrološke osobine ovog plitkog ekosistema i bujan razvoj makrofitske vegetacije od aprila do oktobra, uslovljavaju relativno malu brojnost i biomasu, ali veliki specijski diverzitet fitoplanktona, gde pored silikatnih algi, drugu značajnu komponentu fitoplanktona čine miksotrofne flagelate. Živi svet Zasavice je pod snažnim uticajem sezonskih hidro- meteoroloških prilika u njenom slivu, a biološke interakcije igraju važnu ulogu u dinamici i oblikovanju strukture fitoplanktona. Presudan uticaj temperature na sezonsku dinamiku biomase i strukturu fitoplanktona potvrđuju i analize redundantnosti. Prema parametrima fitoplanktona po Pravilniku Republike Srbije, vrednostima mađarskog Indeksa fitoplanktonskih grupa - Q indeksa i Nemačkog multiparametarskog indeksa fitoplanktona - PSI indeksa, ekološki status Zasavice se može okarakterisati kao dobar. Dodatno, prosečne godišnje vrednosti koncentracije hlorofila a (13,8 µg/L na Lokalitetu 1 i 10,6 µg/L na Lokalitetu 2), ukazuju da je Zasavica eutrofan ekosistem, ali i ekosistem odličnog ekološkog statusa po ovom parametru, što upućuje da trofički status vodenog ekosistema nije izjednačen sa njegovim ekološkim statusom koji zavisi, između ostalog, i od osobina referentnog ekosistema. U epilitskoj zajednici Zasavice identifikovano je ukupno 134 taksona silikatnih algi. U najvećem broju uzoraka Amphora pediculus i Planothidium frequentissimum se pojavljuju kao dominantni ili subdominantni taksoni. Među pripadnicima ove zajednice uočeno je i prisustvo invazivne vrste Diadesmis confervacea. Na osnovu vrednosti IPS dijatomnog indeksa, jedinog relevantnog parametra za procenu ekološkog statusa Zasavice na osnovu bentosnih silikatnih algi po pomenutom Pravilniku Republike Srbije, ekološki status ocenjen je kao dobar. Većina ostalih dijatomnih indeksa pokazuje relativno visok stepen korelacije sa IPS indeksom, pa i njihove vrednosti ukazuju na dobar ekološki status Zasavice. Međutim, izuzetak čine TDI i TID dijatomni indeksi, a njihove vrednosti skreću pažnju na povećanu koncentraciju nutrijenata. Iako algološki parametri upućuju na dobar ekološki status, hemijski parametri ukazuju na loš kvalitet vode, pri čemu je najzaslužniji parametar povećana koncentracija nitratnog jona (prosečna vrednost 11 mg/L na Lokalitetu 1 i 13 mg/L na Lokalitetu 2). Pored toga, koncentracija rastvorenog kiseonika tokom letnjih meseci ukazuje na skoro anoksične uslove (samo 0,24 mg/L u avgustu na Lokalitetu 1), iako prosečne vrednosti od 8,83 mg/L na Lokalitetu 1 i 8,11 mg/L na Lokalitetu 2 upućuju na dobar kvalitet vode. Generalno, algološki parametri Zasavice upućuju na njen dobar ekološki status, ali i na činjenicu da je koncentracija nutrijenata u vodi povišena. Stoga se posebna pažnja mora usmeriti ka smanjenju unosa nutrijenata u ovaj osetljiv ekosistem, ali i održavanju dovoljne količine vode tokom letnje sezone kako bi se zadržao dobar ekološki status Zasavice i trenutno stabilno stanje kojeg karakterišu gust razvoj makrofitske vegetacije i mala gustina fitoplanktona.

A unique ecosystem of Zasavica, the basis of the eponymous Special nature reserve, originated as river ecosystem. Nowadays, however, it represents an oxbow lake or wetland, in ecological terms comparable with shallow lakes, ponds and swamps. To achieve and maintain good ecological status of the wetlands is one of the main aims of Water Framework Directive, in which ecological status is defined as an expression of the quality of the structure and functioning of ecosystem. The ecological status assessment of Serbian waters is regulated through the Rulebook on parameters of ecological and chemical status of surface waters, and quantitative and chemical status of groundwaters. The aim of this PhD thesis was to conduct a comprehensive analysis of cyanobacteria and algae in order to assess the ecological status of Zasavica wetland, using phytoplankton and phytobenthos assemblages as relevant biological quality elements. In addition, metaphyton community was analyzed to support the data on stability of this vulnerable ecosystem. Samples for physical, chemical and biochemical (chlorophyll a concentration) analysis of water quality parameters, as well as samples for quantitative and qualitative analysis of phytoplankton, phytobenthos (epilithic diatoms) and metaphyton communities, were collected monthly from two localities, in the period from December 2012 to November 2013. Further, a standard laboratory methods and formulas for calculating selected indices were used to conduct the analysis. A total of 503 taxa belonging to 8 phyla (Cyanobacteria, Bacillariophyta, Chlorophyta, Chrysophyta, Cryptophyta, Dinophyta, Euglenophyta and Xanthophyta) were identified after the qualitative analysis. Bacillariophyta was the most diverse division, with 170 identified taxa. Regarding Cyanobacteria, out of 50 recorded taxa, 12 were new for the Serbian flora. However, there were 13 cyanobacterial species that are potentially toxic and, among them, three are regarded as invasive in Europe (Cylindrospermopsis raciborskii, Raphidiopsis mediteranea and Sphaerospermopsis aphanizomenoides). Out of 89 observed taxa of the division Euglenophyta, 9 were new records for the Serbian flora. Furthermore, considering that 28 identified taxa of Chrysophyta represent more than a half of the entire diversity of golden algae in Serbia, Zasavica can be characterized as a habitat of exquisite richness concerning the diversity of this division, where as many as 16 new species were recorded. Besides the great species diversity of phytoplankton, a considerable functional diversity, observed through three different classification systems, was also registered. The application of functional phytoplankton classification provides data comparable at a global scale, which facilitates the interpretation of the results obtained in ecological studies of aquatic ecosystems. Sufficient amount of nutrients, geomorphological and hydrological features of this shallow ecosystem, as well as an extensive development of macrophytes from April to October, resulted in a relatively low phytoplankton abundance and biomass, but with high species diversity. With regards to that, it is essential to note that (besides diatoms) mixotrophic flagellates represented the second most important component of the phytoplankton community. In general, life in Zasavica is strongly influenced by hydro- meteorological events occurring within its catchment, while biological interactions play a significant role in the dynamic and structuring of phytoplankton. Temperature is the crucial factor controlling the seasonal dynamic of phytoplankton biomass and structure, which was further confirmed by redundancy analysis. According to the phytoplankton parameters of the Rulebook of Republic of Serbia, values of the Hungarian phytoplankton assemblage index (Q) and German Phyto-See-Index (PSI), ecological status of Zasavica can be characterized as good. In addition, the mean annual values for chlorophyll a concentration (13,8 µg/L and 10,6 µg/L at Localities 1 and 2, respectively) indicate that Zasavica is an eutrophic ecosystem, yet with excellent ecological status. This situation illustrates that trophic state of the aquatic ecosystem is not the same term as ecological status, considering that the latter primarily depends on the characteristics of the reference ecosystem. While observing epilithic community, a total of 134 diatom taxa were recorded. Amphora pediculus and Planothidium frequentissimum appear as the dominant or subdominant taxa in the majority of samples. Among the representatives of this group, an invasive species Diadesmis confervacea was also detected. The values obtained using the IPS diatom index, the only relevant parameter for assessing ecological status of Zasavica based on benthic diatoms according to the above-mentioned Rulebook, indicated a good ecological status. The majority of other diatom indices showed a relatively high correlation with the IPS index, suggesting a good ecological status, as well. The exceptions were TDI and TID diatom indices, and their values implied a high nutrient concentration. Although the algological parameters have suggested a good ecological status of Zasavica, the chemical parameters have characterized its water quality as poor, especially regarding the concentration of nitrate ion, which was very high (mean values for Localities 1 and 2 were 11 mg/L and 13 mg/L, respectively). Furthermore, oxygen levels during the summer months indicated nearly anoxic conditions (only 0,24 mg/L in August at Locality 1), even though the mean values have pointed to a good ecological status (8,83 mg/L for Locality 1 and 8,11 mg/L for Locality 2). In general, the algological parameters have indicated a good ecological status of Zasavica, despite the fact that the level of nutrients in the water was increased. Therefore, it is necessary to act towards decreasing an inflow of nutrients into this vulnerable ecosystem, as well as to maintain optimal water level during the summer season, in order to preserve the good ecological status of Zasavica and its stable state, characterized by the dense development of macrophyte vegetation and low phytoplankton abundance.

Graditeljstvo banja Srbije - 19. i prva polovina 20. veka

Architecture in the spas of Serbia - the 19th and the first half of the 20th century

Polazni cilj ovog rada je istraživanje prirodnih, istorijskih i kulturnih preduslova za nastanak, razvoj i oblikovanje graditeljstva u banjama Srbije u periodu od 1830. do 1940. godine. Dobijeni rezultati omogućavaju određivanje uzajamnih veza i uticaja istorijskih, društvenih, socijalnih i kulturnih prilika i modernizacijskih procesa u državi na ovo graditeljstvo. Sagledavanje razvoja i izgradnje, rasvetljava uloge države i državnih ustanova, organizovanih grupa zainteresovanih građana i privatnih naručilaca u ovom procesu, kao i doprinose pojedinih graditelja na oblikovne karakteristike banjskih kompleksa i građevina. Banjska i klimatska lečilišta i mesta za odmor i oporavak jesu naselja nastajala na prostorima sa posebnim prirodnim uslovima i sa jasno koncipiranim sadržajima. Posle obimnih ispitivanja prirodnih resursa država je od tridesetih godina 19. veka počela da stvara pravni okvir od značaja za uređenje, izgradnju i unapređenje banja, sprovodi njegove odredbe i utiče na glavne smernice razvoja graditeljstva. U 19. i prvoj polovini 20. veka službenici ministarstva u oblasti graditeljstva bili su značajni i ugledni graditelji, talentovane arhitekte različitih generacija, posebnog stilskog i umetničkog izraza. Na osnovu dostupnih izvora, građe i materijala sa terenskih istraživanja rasvetljeni su radovi manje poznatih arhitekata, kao i do sada nepoznata ostvarenja značajnih srpskih arhitekata. U kreiranju stilskih tokova u banjskom graditeljstvu značajnu ulogu imali su zahtevi privatnih naručilaca i lekara, te su i oni predmet analiza. Pored toga, utvrđeni su i oblici prenošenja elemenata graditeljstva srednjoevropskih banja u Srbiju, tokom procesa evropeizacije i modernizacije, koji je pratila ubrzana industrijalizacija, urbanizacija i razvoj saobraćaja. To je donelo nove oblike i pravila društvenog života i nove modele ponašanja građanstva. Kako graditeljstvo banja Srbije do sada nije bilo predmet posebnog, sveobuhvatnijeg i detaljnijeg istraživanja, ovaj rad donosi naučno argumentovano tumačenje njegovog nastanka i razvoja, izdvaja najznačajnije V građevine i vrednosti, kako bi se ocenilo njihovo mesto u ukupnom graditeljstvu posmatranog perioda. Na osnovu sprovedenog istraživanja, proučavanja, valorizacije i istoriografske analize graditeljstva omogućava se utvrđivanje smernica daljeg razvoja i korišćenja banjskog graditeljstva sa stanovišta zaštite kulturnog i prirodnog nasleđa i održivog razvoja. Ovaj rad treba da bude podsticaj i polazište drugim naučnim oblastima za utvrđivanje pravaca razvoja i usmeravanje aktivnosti ka boljem korišćenju prirodnih posebnosti i graditeljstva banja kao ekonomskog potencijala.

The starting goal of the paper is the research on the natural, historical and cultural preconditions for the origins, development and shaping the Serbian spa architecture in the period from 1830 to 1940. The obtained results clear the path for determining the basic reciprocal connections and influences of historical, social and cultural circumstances and modernisation processes in the country on this type of architecture. An insight into its development and construction sheds light on the roles of the state and its agencies, organised groups of stakeholders and private clients in this process, as well as on the impact of certain architects on the spa complexes and buildings shape characteristics. The spa, climatic and health resorts are settlements that originated in the areas of special natural characteristics and with clearly designed facilities. In the 1830s, after conducting extensive research in natural resources, the state started shaping a legal framework for the spas structuring, construction and improvement, then enforcing its provisions and implementing the key guidelines on the architectural development. In the 19th and the first half of the 20th centuries, the Ministry of Building civil servants were important and eminent builders, talented architects of various generations, expressing their particular styles and artistic forms. The available sources and the material gathered during the field investigations brought to light the works of some little-known architects, along with some unknown works of the eminent ones. In addition, private clients’ and physicians’ requirements had their significant role in creating style trends in the spa architecture, so they also became a subject of the research. Furthermore, it has been ascertained how architectural elements from the Central European spas were brought to Serbia during the process of Europeanisation and modernisation, which was followed by a fast paced industrialisation, urbanisation and traffic development. All that brought new forms and rules of social life and new models of civil conduct. Since the architecture of the Serbian spas has not been a subject of any particular, extensive and detailed research so far, this paper now brings forth a scholarly well-documented interpretation of its origins and development and VII accentuates the most important buildings and values so as to facilitate the valorisation of its place in the overall architecture of the studied period. Therefore, the conducted investigations, research, valorisation and historiographic analysis in architecture are the grounds for establishing the guidelines for further development and utilisation of the spa architecture from the aspect of cultural and natural heritage conservation and sustainable development. This paper aims at being an incentive and a starting point to other academic fields for establishing directions of development and activities towards a better use of the spa natural resources and architectural properties as an economic potential.

Mikro RNK-21 kao indikator invazivnosti karcinoma dojke kod pacijenata sa različitim statusom estrogenskog i progesteronskog receptora

MicroRNA-21 as an indicator of breast carcinoma invasiveness in patients with different estrogen and progesterone receptor status.

Karcinomi dojke su izuzetno heterogeni tumori koji se na osnovu sposbonosti invazivnosti dele na neinvazivne, invazivne asocirane sa neinvazivnim i invazivne. Najznačajniji standardni dijagnostički i prognostički parametri karcinoma dojke su veličina tumora, starost pacijentkinje, status regionalnih limfnih čvorova, gradus tumora, status estrogenskog receptora (ER), progesteronskog receptora (PR), receptora za humani epidermalni faktor rasta 2 (Her-2) i status proliferativnog indeksa (Ki-67). Male nekodirajuće mikro RNK igraju važnu ulogu u regulaciji velikog broja bioloških procesa, a poremećaji u njihovoj ekspresiji detektovani su u procesima nastanka i progresije malignih tumora. Cilj ove disertacije je da se ispita nivo i značaj ekspresije onkogene mikro RNK-21 (miR-21) u tumorskom tkivu dojke. Imunohistohemijski je analiziran nivo ekspresije proteina ER, PR, Her-2 i Ki-67 u neizmenjenom i tumorskom tkivu dojke dok je nivo ekspresije miR-21 određivan lančanom rekacijom polimeraze kojoj prethodi reverzna transkripcija korišćenjem prajmera sa petljom. Razlike u nivoima ekspresije miR-21 u uzorcima neinvazivnih i invazivnih karcinoma dojke sa in situ koponentom ukazuju na moguću biološku ulogu miR-21 u procesima odgovornim za invazivnost karcinoma dojke, odnosno da nvo ekspresije molekula mikro RNK-21 predstavlja potencijani biološki marker invazivnosti karcinoma dojke. Rezultat da u analiziranoj grupi čistih invazivnih karcinoma dojke postoji visok stepen pozitivne korelacije između nivoa ekspresije molekula mikro RNK-21 sa nivoom ekspresije proteina ER i PR, ukazuje na to da nivo ekspresije molekula mikro RNK-21 može predstavljati novi nestandardni dijagnostički i prognostički parametar karcinoma dojke.

Breast cancer is a complex and heterogeneous group of tumors. According to their ability to invade, breast cancers are divided in three major types: non-invasive, invasive carcinoma of the breast associated with non-invasive and invasive breast carcinomas. The most significant standard diagnostic and prognostic parameters of the breast carcinomas are size of a tumor, age at diagnosis, regional lymph-node status, tumor grade, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor 2 receptor status and proliferative index Ki-67 status. Small non-coding microRNAs play very important part in the regulation of great number of biological processes. Deregulation in their expression are detected in the process of malignant tumor formation and progression. Main goal of this dissertation was to investigate expression levels of microRNA-21 (miR-21) in the tumorous breast tissue and its potential role in breast tumor formation and invassion. Expression levels of ER, PR, Her-2 and Ki-67 proteins were detected by immunohistochemistry in non-transformed and tumorous tissue of the breast. MicroRNA-21 expression levels were determined by the reaction of Reverse Transcription and Stem-Loop Polymerase Chain Reaction. The differences in expression levels of miR-21 among non-invasive, invasive breast carcinoma associated with non-invasive and invasive breast carcinomas emphasizes potential biological role of miR-21 in the proces of breast cancer invasiveness and indicates that expression levels of microRNA-21 might represent potential biomarker of invasiveness in the breast carcinomas. We have found that there was highly positive correlation in miR- 21 expression levels with ER and PR protein expression levels, which indicates that miR- 21 expression levels might be used as novel non-standard parameter for breast cancer diagnosis and prognosis.

Fitocenološko-ekološka studija močvarne vegetacije (Phragmitetea communis R. Th. et Preising 1942) centralnog Balkana

A phytosociological-ecological study of marshland vegetation (Phragmitetea communis R. TX. et Preising 1942) in the central Balkans

Prvi zapisi o florističkom sastavu, ekologiji i distribuciji močvarne vegetacije centralnog Balkana postali su dostupni naučnoj javnosti početkom XX veka. Višedecenijsko razdoblje, od prvih godina prošlog veka do danas, obeležila su sporadična i fragmentarna proučavanja ovog vegetacijskog tipa. Mnogobrojna staništa obrasla zajednicama trski i visokih oštrica, rasprostranjena u nizijskim, brdskim i planinskim predelima centralnog Balkana, ostala su neistražena kako sa fitocenološkog, tako i ekološkog aspekta. U cilju obogaćivanja saznanja o kvalitativnom i kvantitativnom sastavu vrsta, distribuciji, sezonskoj varijabilnosti i ekologiji močvarnih fitocenoza centralnog Balkana izvršeno je istraživanje, do sada, neproučavanih staništa smeštenih na teritoriji slivnog područja Južne Morave. Realizacijom pomenutog istraživanja stvorena je adekvatna osnova za postavljanje i realizaciju glavnih ciljeva ove studije, među kojima su: utvrđivanje sintaksonomskih odnosa u okviru vegetacijske klase Phragmitetea communis, razmatranje mogućnosti predlaganja uniformnog vremena uzorkovanja močvarnih fitocenoza, definisanje bioindikatorskog potencijala pojedinih makrofita (statistički značajnih indikatorskih vrsta proučavanih fitocenoza) na osnovu prikupljenih informacija o njihovim ekološkim afinitetima u pogledu fizičko-hemijskih osobina vode i supstrata. Primenom hijerarhijske klasifikacione analize na setu podataka koji objedinjuje, do sada, nepublikovane fitocenološke snimke zabeležene na proučavanom području i fitocenološke snimke preuzete iz literaturnih izvora ustanovljeno je da močvarnu vegetaciju centralnog Balkana odlikuje izuzetna raznovrsnost asocijacija. Na osnovu razlika u kvalitativnom i kvantitativnom sastavu taksona, prepoznato je i opisano 27 asocijacija, Typhetum angustifoliae, Typhetum latifoliae, Typhetum domingensis, Phragmitetum australis, Schoenoplectetum lacustris, Glycerietum maximae, Equisetetum limosi, Glycerietum fluitantis, Glycerietum notatae, Sparganietum erecti, Oenantho aquaticae-Rorippetum amphibiae, Phalaridetum arundinaceae, Eleocharietum palustris, Butometum umbellati, Bolboschoenetum glauci, Caricetum gracillis, Caricetum ripariae, Caricetum vulpinae, Caricetum rostrato-vesicariae, Cyperetum longi, Caricetum elatae, Cladietum marisci, Caricetum paniculatae, Caricetum acutiformis, Bolboschoenetum maritimi continentale, Schoenoplectetum tabernaemontani i Oenantho fistulosae-Beckmannietum eruciformis, u okviru 5 vegetacijskih redova i 7 sveza vegetacijske klase Phragmitetea communis. Detaljne informacije o florističkoj sličnosti sastojina, njihovoj ekologiji i distribuciji prikazane su za svaku asocijaciju ponaosob u glavnom delu doktorske disertacije. Permutacionim multivarijantnim analizama varijanse ustanovljeno je da od 12 proučavanih zajednica (Bolboschoenetum maritimi continentale, Caricetum ripariae, Eleocharietum palustris, Glycerietum maximae, Phalaridetum arundinaceae, Phragmitetum australis, Schoenoplectetum lacustris, Schoenoplectetum tabernaemontani, Sparganietum erecti, Typhetum angustifoliae, Typhetum domingensis i Typhetum latifoliae) samo dve fitocenoze, Bolboschoenetum maritimi continentale i Eleocharietum palustris, pokazuju statistički značajne sezonske promene kvalitativnog i kvantitativnog sastava vrsta i životnih formi. Neparametarskim Friedman-ovim testom utvrđeno je postojanje statistički značajnih sezonskih promena α-diverziteta u zajednicama Typhetum angustifoliae i Typhetum domingensis. Osim toga, istom statističkom analizom konstatovano je da se broj vrsta i vrednosti α-diverziteta zajednica Glycerietum maximae, Phragmitetum australis i Schoenoplectetum lacustris statistički značajno menjaju tokom vegetacione sezone. Sezonska varijabilnost močvarnih zajednica ne ogleda se samo u promenljivom kvalitativnom i kvantitativnom sastavu vrsta, životnih formi, broja vrsta i α-diverziteta, već i u promenljivom sastavu statistički značajnih indikatorskih vrsta, a navedeni oblik sezonske varijabilnosti karakteriše sledeće fitocenoze: Schoenoplectetum lacustris, Sparganietum erecti i Typhetum angustifoliae. U skladu sa rezultatima primenjenih statističkih testova zapaženo je da sezonsku dinamiku močvarnih zajednica iniciraju promene u vodnom režimu njihovih staništa. Nepredvidivost trajanja i smenjivanja pojedinih ekofaza tokom vegetacione sezone ograničavaju mogućnost predikcije sezonske promenljivosti močvarnih zajednica, a samim tim i predlaganja optimalnog vremena njihovog uzorkovanja. Shodno tome, u budućim fitocenološkim studijama bi trebalo utvrditi posledice uključivanja sezonski heterogenih fitocenoloških snimaka u iste statističke analize na definisanje vegetacijskih jedinica i donošenje sintaksonomskih zaključaka. Ekološki afiniteti i bioindikatorski potencijali statistički značajnih indikatorskih vrsta proučavanih fitocenoza procenjeneni su na osnovu oblika kriva odgovora i vrednosti njihovih kardinalnih tačaka. Vrste Bolboschoenus maritimus i Scirpus lacustris subsp. tabernaemontani prepoznate su kao validni bioindikatori blago zaslanjenih, alkalnih staništa na kojima je koncentracija SO4 2- u vodi visoka. Ekološke valence pomenutih vrsta samo su delimično preklopljene jer vrsti Bolboschoenus maritimus, u odnosu na vrstu Scirpus lacustris subsp. tabernaemontani, pogoduju slanija i alkalnija staništa koja su siromašnija sulfatima, a bogatija azotom, fosforom, kalijumom i hloridima. Inače, stepen saliniteta, kiselosti i sadržaj nutrijenata na močvarnim staništima ubrajaju se među ključne faktore ekološke diverzifikacije vrsta roda Bolboschoenus. Vrsta Bolboschoenus glaucus, za razliku od vrste Bolboschoenus maritimus, preferira nezaslanjena, blago alkalna staništa na kojima je sadržaj nutrijenata relativno nizak. Staništa na kojima je voda relativno duboka, blago kisela, siromašna SO4 2- (0.30 mg/l), a supstrat sadrži malu količinu K2O (6.80 mg/100g sedimenta) i ima nizak stepen elektroprovodljivosti pogoduju vrstama Typha angustifolia, Sparganium erectum i Typha latifolia. Primenom Huisman-Olff-Fresco (HOF) modela konstatovano je da su močvarna staništa okarakterisana niskim ili osrednjim sadržajem NO3 -, NH4 +, PO4 3- i SO4 2- u vodi pogodna za razvoj i rast vrsta Carex riparia, Eleocharis palustris, Phalaris arundinacea, Scirpus lacustris i Stachys palustris. Rezultatima ove studije potvrđeno je da vrsta Phragmites australis ima nisku indikativnu vrednost i regionalni bioindikatorski značaj. Višemesečnim monitoringom močvarnih ekosistema centralnog Balkana obogaćena su saznanja o ekološkim afinitetima pojedinih makrofita, a time je napravljen prvi korak u formiranju nacionalne baze podataka o univarijantim odgovorima hidrofita. Prikupljanjem informacija o univarijantnim odgovorima vrsta u odnosu na različite sredinske varijable postavlja se dobra osnova za korigovanje neadekvatno definisanih vrednosti ekoloških indeksa, definisanje indikatorskih vrednosti u odnosu na sredinske varijable koje u postojećim indikatorskim sistemima nisu razmatrane i realizaciju aktivnosti koje su usmerene ka obnovi, unapređivanju i zaštiti močvarnih ekosistema.

The first records of the floristic composition, ecology and distribution of marshland vegetation in the Central Balkans became available for the scientific community at the beginning of the 20th century. From then until the present day there has only been a sporadic and fragmentary study of this vegetation type. Numerous habitats covered by reed and tall sedge communities, spread on the plain, hilly and mountainous terrain of the Central Balkan Peninsula, remain unexplored from their phytocoenological and ecological aspects. In order to enrich the current knowledge of the qualitative and quantitative species composition, distribution, seasonal variation and ecology of the marshland communities of the Central Balkans, research was conducted on the unstudied habitats located in the South Morava river basin. To carry out the research, an adequate basis for setting up and implementing the main goals of this study was created, including: determining the syntaxonomical relationships within the vegetation class Phragmitetea communis; considering the possibility of proposing a uniform time for sampling marshland communities; and defining the bioindicator potential of some macrophytes (statistically significant indicator species in these communities) based on information collected about their ecological preferences in terms of the physicоchemical properties of the water and substrate. By applying hierarchical classification analysis to a data set that combines so far unpublished relevés recorded in the study area and relevés taken from literature sources, it was established that the marshland vegetation of the Central Balkans is characterized by an exceptional variety of associations. On the basis of differences in the qualitative and quantitative composition of the species, 27 associations were recognized and described, Typhetum angustifoliae, Typhetum latifoliae, Typhetum domingensis, Phragmitetum australis, Schoenoplectetum lacustris, Glycerietum maximae, Equisetetum limosi, Glycerietum fluitantis, Glycerietum notatae, Sparganietum erecti, Oenantho aquaticae-Rorippetum amphibiae, Phalaridetum arundinaceae, Eleocharietum palustris, Butometum umbellati, Bolboschoenetum glauci, Caricetum gracillis, Caricetum ripariae, Caricetum vulpinae, Caricetum rostrato-vesicariae, Cyperetum longi, Caricetum elatae, Cladietum marisci, Caricetum paniculatae, Caricetum acutiformis, Bolboschoenetum maritimi continentale, Schoenoplectetum tabernaemontani and Oenantho fistulosae-Beckmannietum eruciformis, within 5 orders and 7 alliances in the vegetation class Phragmitetea communis. Detailed information on the ecology, distribution and floristic similarities between the stands studied is given for each association separately in the main part of the doctoral dissertation. The results of the permutational multivariate analysis of variance show that of the twelve communities studied (Bolboschoenetum maritimi continentale, Caricetum ripariae, Eleocharietum palustris, Glycerietum maximae, Phalaridetum arundinaceae, Phragmitetum australis, Schoenoplectetum lacustris, Schoenoplectetum tabernaemontani, Sparganietum erecti, Typhetum angustifoliae, Typhetum domingensis and Typhetum latifoliae) only two communities, Bolboschoenetum maritimi continentale and Eleocharietum palustris, show statistically significant seasonal variability concerning the qualitative and quantitative composition of the species and life forms. Using the non-parametric Friedman test, it was established that the communities Typhetum angustifoliae and Typhetum domingensis show statistically significant seasonal changes of α-diversity. In addition, the same statistical analysis revealed that the number of species and the value of α-diversity for the communities Glycerietum maximae, Phragmitetum australis and Schoenoplectetum lacustris significantly change during the growing season. The seasonal variability of marshland communities is not only seen in the rapidly changing qualitative and quantitative composition of the species, life forms, number of species and α-diversity, but also in the variable composition of statistically significant indicator species. This seasonal variability was seen in the following communities: Schoenoplectetum lacustris, Sparganietum erecti and Typhetum angustifoliae. In accordance with the results of the statistical tests applied, it was noticed that the seasonal dynamics of the marshland communities depend on the annual changes in the water regime. The unpredictability of the duration and the alternation of some ecophases during the growing season limit the possibility of predicting the seasonal variability of marshland communities, and consequently proposing the optimal timing for their sampling. Accordingly, evaluating the effects of using seasonally heterogeneous relevés of marshland vegetation in the same statistical analyses in order to define the vegetation units and to bring syntaxonomical conclusions should be an aim for further phytosociological studies. The ecological affinities and bioindicator potential of statistically significant indicator species in the communities studied are based on shape of species response curves and the value of their cardinal points. Bolboschoenus maritimus and Scirpus lacustris subsp. tabernaemontani were recognized as valid biological indicators of mildly saline, alkaline habitats in which the concentration of SO4 2- in the water is high. Their ecological valences are only partially overlapped. Bolboschoenus maritimus, in relation to Scirpus lacustris subsp. tabernaemontani, prefers saltier, more alkaline habitats that are poorer in sulfates, and enriched with nitrogen, phosphorus, potassium and chloride. Otherwise, the degree of salinity, acidity and nutrient content of the marshland habitats are the key factors in the ecological diversification of the Bolboschoenus species. Bolboschoenus glaucus, unlike Bolboschoenus maritimus, prefers unsalted, slightly alkaline habitats where the nutrient content is relatively low. Habitats characterized with relatively deep, slightly acidic water which is poor in SO4 2- (0.30 mg/l) and substrate that contains a small amount of K2O (6.80 mg/100g sediment) and has a low level of conductivity are suitable for the optimal development of Typha angustifolia, Sparganium erectum and Typha latifolia. Using the Huisman-Olff-Fresco (HOF) models, it was established that marshland habitats with a low or mediocre content of NO3 -, NH4 +, PO4 3- and SO4 2- in the water are suitable for the development and growth of species Carex riparia, Eleocharis palustris, Phalaris arundinacea, Scirpus lacustris and Stachys palustris. Further, the results of this study confirmed that the species Phragmites australis has a low indicative value and regional bioindicator potential. By continuously monitoring the marshland ecosystems of the Central Balkan Peninsula the existing knowledge on the ecological preferences of some macrophytes has been enriched, and thus the first step has been made towards creating a national database on the univariate response of hydrophytes. Collecting information on the univariate responses of species in relation to different environmental variables sets up an appropriate basis for the following: correcting inadequately defined ecological index values; defining indicator values in regard to environmental variables that have not yet been considered in the current indicator systems; and implementing activities aimed at the reconstruction, promotion and protection of marshland ecosystems.

Uloga evolucionih interakcija između intracelularnog endosimbionta (Wolbachia) i fitoplazme (Flavescence dorée) u promenama komponenti adaptivne vrednosti i pravcima evolucije mitohondrijske DNK u prirodnim populacijama Dictyophara europaea

The role of evolutionary interactions between intracellular endosymbiont (Wolbachia) and phytoplasma (Flavescence dorée) on fitness components and evolution of mitochondrial DNA in natural populations Dictyophara europaea

Dictyophara europaea je široko rasprostranjena i ekonomski značajna cikada koja kao alternativni vektor Flavescence dorée (FD) fitoplazme, učestvuje u prenošenju bolesti u vinogradima Evrope. Sprovedena epidemiološka istraživanja su ukazala na postojanje populacija D. europaea sa veoma visokom stopom FD-infekcije, Wolbachia-inficiranih populacija sa niskom stopom FD-infekcije (DeWo+) i odsustvo Wolbachia u populacijama sa visokom stopom FD-infekcije (DeWo-). Ispitivano je nekoliko mogućih uzroka raličite stope infekcije vektora: i) populaciono-genetičke karakteristike D. europaea i korelacija sa Wolbachia infekcijom, ii) efekat Wolbachia na komponente adaptivne vrednosti laboratorijske kolonije DeWo+, i iii) stopa inficiranosti prirodnog biljnog rezervoara FD fitoplazme i razlike u genotipovima fitoplazme u nisko- i visoko- inficiranim populacijama. Genotipizacijom mitohondrijskog COI gena evidentiran je smanjen genetički diverzitet DeWo+ u poređenju sa DeWo- populacijama i različita evolucija fiksiranih haplotipova. Multilokusnom genotipizacijom Wolbachia izolata identifikovan je isti soj, wEur, u svim DeWo+ populacijama. Diverzitet FD fitoplazme nije bio u korelaciji sa različitom stopom inficiranosti vektora. Utvrđeno je da Wolbachia-infekcija nema direktnog efekta na adaptivnu vrednost D. europaea. Podaci iz prirodnih populacija D. europaea ukazuju na kompeticiju Wolbachia i FD fitoplazme: populacije koje su u niskoj stopi inficirane FD fitoplazmom su Wolbachia-inficirane i vice versa.

Dictyophara europaea is widespread planthopper of economic importance, acting as an alternative vector of the Flavescence dorée (FD) phytoplasma disease of grapevine in European vineyards. Epidemiological studies have revealed D. europaea populations that are heavily FD-infected, as well as populations with low FD-infection rates that are naturally infected with Wolbachia (DeWo+), and highly FD-infected populations in the absence of Wolbachia (DeWo-).Several possible causes of differences in vector infection rates were examined: i) population genetic characteristics of D. europaea and their correlation with Wolbachia infection, ii) Wolbachia effects on fitness components of DeWo+ laboratory colony, and iii) the rate of reservoir plant FD-infection and differences in FD genotypes harboured by low and high infected vector populations. Genotyping of mitochondrial COI loci showed lower genetic diversity level in DeWo+ than in DeWo- populations of and divergent evolution of fixed haplotypes. Multilocus genotyping of Wolbachia revealed that all DeWo+ populations were infected with the same strain (i.e. wEur). Genetically diversified FD phytoplasma had no direct influence on vector infection rates. No evidence of direct fitness effects on D. europaea was registered with regard to Wolbachia infection status. Field data and the observed negative correlation between FD-infection and Wolbachia-infection rates, indicate that Wolbachia compete with FD phytoplasma within the host insect D. europaea.

Ispitivanje uloge molekula vanćelijskog matriksa tenascina-C u regulaciji strukturne i funkcionalne plastičnosti malog mozga i oblikovanju ponašanja miša nakon izlaganja uslovima obogaćene sredine

Investigation of the role of the extracellular matrix molecule Tenascin-C in the regulation of structural and functional plasticity of the cerebellum and in shaping of the behavior of the mouse after exposure to enriched environment conditions

Tenascin-C (TnC) je glikoprotein prisutan u vanćelijskom matriksu (VĆM) različitih tkiva kičmenjaka tokom razvića, gde je uključen u regulaciju ćelijskog rasta, migracije i adhezije preko aktivacije različitih unutarćelijskih signalnih puteva. Ekspresija TnC je značajno smanjenja u adultnom organizmu, međutim ostaje prisutna samo u određenim perifernim tkivima, kao i u delovima centralnog nervnog sistema koji zadržavaju visok nivo plastičnosti, kao što je mali mozak, gde je uočena njegova uloga u modulaciji sinaptičkih funkcija. Obogaćena sredina (OS) oblikuje mnoge aspekte ponašanja i stoga može da posluži kao pristup za proučavanje neuronalne plastičnosti u adultnom organizmu. Izlaganje OS dovodi do brojnih promena na molekularnom i ćelijskom nivou u različitim regionima mozga uključujući mali mozak. Cilj ove studije je bio da se ispita uloga TnC u modulaciji strukturne plastičnosti malog mozga indukovane OS praćenjem distribucije perineuronalnih mreža (PNM), promene u veličini i gustini ekscitatornih i inhibitornih presinaptičkih završetaka, kao i aktivnosti glavnih enzima koji vrše razgradnju VĆM, MMP-2 i MMP-9. Takođe, ova studija je imala za cilj da ispita kako nedostatak TnC interaguje sa spoljašnjom sredinom u oblikovanju različitih domena ponašanja povezanih sa funkcijom malog mozga. U te svrhe, miševi sa nedostatkom TnC (TnC-/-) ili MMP-9 (MMP-9-/-) i odgovarajući kontrolni miševi starosti 3 nedelje su izlagani standardnim uslovima gajenja (SS) ili OS 4 ili 8 nedelja. Ova studija je pokazala da izlaganje OS u toku 8 nedelja dovodi do smanjenja intenziteta bojenja na PNM, kao i smanjenja u veličini GABAergičkih i povećanja broja i veličine glutamatergičkih sinaptičkih završetaka u kontrolnim miševima. Nasuprot tome, TnC-/- miševi su pokazali smanjen intenzitet bojenja na PNM u poređenju sa kontrolnim životinjama gajenim u SS, dok njihovo izlaganje OS nije dovelo da smanjenja, već do blagog povećanja intenziteta PNM. Pored toga, OS nije uticala na gustinu dva tipa sinaptičkih završetaka kod TnC-/- miševa, dok je veličina inhibitornih, ali ne i ekscitatornih, sinaptičkih završetaka bila povećana. Takođe, merenjem aktivnosti MMP-9, kao i praćenjem njene kolokalizacije sa PNM i presinaptičkim markerima, pokazano je da MMP-9, ali ne i MMP-2, utiče na remodelovanje PNM i sinaptičku plastičnost u malom mozgu u vremenskom periodu između 4. i 8. nedelje izlaganja OS. Ovi nalazi su dodatno potvrđeni rezultatima dobijenim na MMP-9-/- miševima. Za procenu ponašajnog fenotipa TnC-/- miševa korišćena je baterija testova nakon izlaganja ovih životinja SS i OS u trajanju od 8 nedelja. Ova studija pokazuje da TnC-/- miševi gajeni u SS ispoljavaju spontanu noćnu hiperaktivnost, kao i slabu senzomotornu koordinaciju i sporo plivanje u Morisovom vodenom lavirintu. Međutim, izlaganje ovih životinja OS je dovelo do ukidanja hiperlokomocije, brže habituacije ovih životinja na novu sredinu, poboljšanja snage prednjih ekstremiteta i delimičnog poboljšanja koordinacije pokreta, dok je sposobnost plivanja ostala poremećena. S druge strane, nedostatak TnC je umanjio pozitivne efekte OS na sposobnost učenja i pamćenja, kao i anksiolitičke efekte OS na smanjivanje nivoa akrofobije. Ova studija pokazuje da je TnC uključen u regulaciju procesa strukturne plastičnosti u malom mozgu, kao i da su interakcije između TnC i MMP-9 neophodne za odigravanje ovih procesa. Takođe, ova studija proširuje postojeće znanje o ponašajnom fenotipu TnC-/- miševa i otkriva da efekat genetičke osnove na ponašajne odgovore može biti izmenjen izlaganjem životinja stimulativnoj sredini.

Tenascin-C (TnC) is a glycoprotein present in the extracellular matrix (ECM) of a variety of vertebrate tissues during development, where it plays multiple roles in cell growth, migration and adhesion by activating diverse intracellular signaling pathways. Expression of TnC is markedly downregulated in the adulthood, persisting in a few peripheral structures and in central nervous system areas known to retain high degree of plasticity, such as the cerebellum, where it is involved in the modulation of synaptic functions. Enriched environment (EE) shapes many aspects of behavior and may, therefore, serve as a paradigm to study neuronal plasticity in the adult. Exposure to EE leads to numerous changes at the molecular and cellular levels, which target various brain regions including the cerebellum. The aim of present study was to examine the role of TnC in the modulation of cerebellar structural plasticity induced by the exposure to EE by following the appearance of perineuronal nets (PNN), changes in size and density of excitatory and inhibitory synaptic terminals, and the activity of major ECM degrading enzymes, MMP- 2 and MMP-9. Furthermore, the present study aimed to examine how TnC deficiency interacts with the environment in shaping different behavioral domains associated with the cerebellum. To this end, 3-week-old mice lacking TnC (TnC-/-) or MMP-9 (MMP- 9-/-) and corresponding wild-type mice were exposed to standard conditions (SC) or an enriched environment (EE) for 4 or 8 weeks. The present study shows that 8 weeks of exposure to EE leads to reduced staining of PNN, reduction in the size of GABAergic and increase in the number and size of glutamatergic synaptic terminals in wild-type mice. Conversely, TnC-/- mice showed reduced staining of PNNs compared to wild-type mice maintained under standard conditions, while exposure to EE did not further reduce, but even slightly increased PNN staining. EE did not affect the densities of the two types of synaptic terminals in TnC-/- mice, while the size of inhibitory, but not of excitatory synaptic terminals was increased. In the time frame of 4-8 weeks, MMP-9, but not MMP-2, was observed to influence PNN remodeling and cerebellar synaptic plasticity as revealed by measurement of MMP-9 activity and colocalization with PNNs and synaptic markers. These findings were supported by observations on MMP-9-/- mice. To asses the behavioral phenotype of TnC-/- mice a comprehensive battery of tests was used after post-weaning housing animals in SC or EE for 8 weeks. The present study shows that when housed in SC, TnC-/- mice exhibited spontaneous nocturnal hyperactivity, as well as poor sensorimotor coordination and low swimming velocity in the Morris water maze. However, housing of TnC-/- mice in EE abolished hyperlocomotion, led to faster habituation to novel environment, strengthened the grasp of fore limbs and partially improved movement coordination, while the swimming ability remained deficient. Conversely, TnC deficiency attenuated both the beneficial effects of EE on learning/memory capacity and the anxiolytic effect of EE in reducing the level of acrophobia. The present study shows that TnC contributes to the regulation of structural plasticity in the cerebellum and that interactions between TnC and MMP-9 are important for these processes to occur. Furthermore, this study expands the existing knowledge about the phenotype associated with TnC deficiency, and reveals that the effect of genetic background on the behavioral response could be altered by housing in a stimulating environment.

In vitro propagacija i biološka aktivnost etarskog ulja i metanolnih ekstrakata povratiča (Tanacetum vulgare L.)

In vitro propagation and biological activity of essential oil and methanol extracts of tansy (Tanacetum vulgare L.)

Rodu Tanacetum pripadaju mnoge aromatične, lekovite i ukrasne vrste bogate biološki aktivnim sekundarnim metabolitima. Tanacetum vulgare L. (syn. Chrysanthemum vulgare L., povratič, vratiželja, konopljika) je višegodišnja, zeljasta biljka, poreklom iz Evrope i centralne Azije koja se obično sreće duž puteva, pruga, pašnjaka i polja ali i priobalnih područja. U Srbiji je ova vrsta deo sinurbane (ruderalne) flore. Predmet istraživanja ove disertacije bila ja fitohemijska karakterizacija sastava i bioloških aktivnosti etarskog ulja i metanolnih ekstrakata ove vrste. In vitro kultura T. vulgare je uspešno uspostavljena iz semena biljaka sakupljenih u prirodi. Kultura izdanaka je uspešno održavana i multiplikovana na MS hranljivoj podlozi uz dodatak 6-benziaminopurina (BAP) a prilikom ožiljavanja BAP je zamenjen indolsirćetnom kiselinom (IBA). In vitro kultura korenova održavana je u tečnoj MS hranljivoj podlozi sa dodatkom IBA koja je uticala na povećanje biomase gajenih korenova tokom subkultura. Histoločkom analizom sekretornih struktura in vitro gajenih izdanaka utvrđeno je da su na listovima i stablu prisutne biserijatne glandularne trihome sa sekrecionim sadržajem u kojem su detektovani lipidi, terpeni i alkaloidi. Hemijska analiza metanolnih ekstrakata herbe i korena T. vulgare ukazala je na to da su ekstrakti biljaka gajenih in vitro kvalitativno siromašniji u odnosu na metanolne ekstrakte biljaka iz prirode ali su se ekstrakti biljaka in vitro odlikovali višestruko većom zastupljenošću pojedinih jedinjenja, pre svih 3,5-O-dikafeoilhininskom kiselinom. Najzastupljenija jedinjenja su fenolne kiseline iz grupe derivata cimetne kiseline i to: neohlorogena, 3,5-O-dikafeoilhininska i dikafeoilhininska kiselina. Pored fenolnih kiselina detektovano je i 17 flavonoida. Relativni udeo fenolnih kiselina i flavonoida se razlikovao u zavisnosti od dela biljke korišćenog za pripremanje matanolnog ekstrakta. Uporedna analiza fitohemijskog sastava isparljivih komponenti etarskog ulja T. vugare sakupljenog sa prirodnog staništa i izdanaka gajenih in vitro pokazala je takođe velike razlike u zastupljenosti pojedinačnih ali i čitavih grupa jedinjenja. GC/MS analizom je pokazano da su kod biljaka iz prirode dominantne komponente u etarskom ulju pripadale grupi oksidovanih monoterpena, sa najzastupljenijim jedinjenjima trans-hrisantenil acetatom , trans-hrizantenolom , trans-tujonom i cis-tujonom. Sa druge strane, etarsko ulje in vitro gajenih biljaka T. vulgare karakterisalo se ujednačenim prisustvom monoterpena i seskviterpena. Svi monoterpeni identifikovani kod in vitro biljaka bili su prisutni i u etarskom ulju biljaka iz prirode dok su seskviterpenska jedinjenja bila daleko zastupljenija i raznovrsnija kod in vitro biljaka. Za analizu bioloških aktivnosti jedinjenja sekundarnog metabolizma T. vulgare korišćeni su etarsko ulje i metanolni ekstrakti dobijeni iz biljaka sakupljenih iz prirode. Rezultati su pokazali da svi analizirani metanolni ekstrakti ispoljavaju značajnu antioksidativnu aktivnost koja je korelirala sa sadržajem ukupnih fenola u ekstraktima. Metanolni ekstrakt korena posedovao je najveći antioksidativni kapacitet a ujedno je sadržavao i najviše ukupnih fenola, sa najvećom relativnom zastupljenošću neohlorogene, 3,5-O- dikafeoilhininske i dikafeoilhininske kiseline u odnosu na ostale ekstrakte. Etarsko ulje i metanolni ekstrakti cveta, lista, stabljike i korena T. vulgare ispoljili su i značajnu antimikrobnu aktivnost testiranu na 8 bakterijskih i 6 vrsta mikromiceta. Etarsko ulje T. vulgare ispoljilo je jako antimikrobno dejstvo prema većini ispitivanih bakterijskih (5 od analiziranih 8) i svim vrstama mikromiceta. S druge strane, metanolni ekstrakti korena, lista, cveta i stabljike T. vulgare ispoljili su jaku aktivnost na sve testirane vrste bakterija i većinu mikromiceta. U većini slučajeva efekti testiranih ekstrakata su bili na nivou efekata referentnih antibiotika i antimikotika korišćenih kao pozitivne kontrole, te se etarsko ulje i metanolni ekstrakti T. vulgare mogu preporučiti u prevenciji i lečenju infekcija izazvanih vrstama prema kojima je pokazana visoka aktivnost ali i kao agensi u konzerviranju i očuvanju kvaliteta namirnica u prehrambenoj industriji. U daljem radu ispitivan je citotoksični potencijal etarskog ulja i metanolnih ekstrakata na maligne ćelijske linije humanog adenokarcinoma cerviksa (HeLa). Radi utvrđivanja nivoa selektivnog efekta ispitivana je i aktivnost na zdrave humane ćelijske linije fetalnih fibroblasta pluća (MRC-5). Rezultati in vitro testa su pokazali da je etarsko ulje ispoljilo najslabiju citotoksičnost dok su metanolni ekstrakti stabla i korena pokazali slabije citotoksično delovanje na ciljne ćelije u odnosu na metanolne ekstrakte lista i cveta. Nakon tretmana metanolnim ekstraktima lista i cveta kod većine HeLa ćelija došlo je do gubitka adhezije, skupljanja membrane i zaokrugljivanja ćelija što je ukazivalo na ćelijsku smrt. Praćenje efekta etarskog ulja T. vulgare na larve gubara (Lymantria dispar L.) pokazalo je da etarsko ulje T. vulgare nije ispoljilo akutnu toksičnost na gusenice drugog stadijuma razvića ali je izazvalo odlaganje presvlačenja i značajno smanjenje procenta presvučenih gusenica. Ingestija etarskog ulja uticala je i na druge parametre rastenja i razvića posmatrane kod gusenica četvrtog stupnja razvića. Etarsko ulje je smanjilo dnevni prirast mase gusenica četvrtog stupnja kao i brzinu konzumacije hrane. Iako je etarsko ulje uticalo i na smanjenje konverzije svarene hrane u biomasu gusenica, to smanjenje se nije pokazalo kao statistički značajno pa se može zaključiti da je time smanjen negativni efekat ingestije etarskog ulja i delom kompenzovan smanjeni unos hrane izazvan prisustvom etarskog ulja. U cilju ispitivanja potencijala etarskog ulja T. vulgare da indukuje mehanizme odbrane in vitro gajenih biljaka krompira koje su rasle u atmosferi ispunjenoj isparljivim komponentama ovog ulja, analizirana je ekspresija 4 gena uključena u mehanizme odbrane protiv patogena i herbivornih insekata. Prisustvo etarskog ulja izazvalo je intenzivnu indukciju dva analizirana PR (eng. pathogenesis-related) gena, PR-2 i PR-5. Prve promene u ekspresiji uočene su nakon 8 sati od izlaganja, a maksimum ekspresije od oko 50 puta zabeležen je za PR-2 gen nakon 12 h. Na osnovu preliminarne PTR-MS analize i merenja zastupljenosti pojedinačnih jedinjenja etarskog ulja u atmosferi teglica u kojima je bio krompir dominantna jedinjenja su bila monoterpenski ugljovodonici α-pinen i p-cimen. Pored ovih jedinjenja i artemizia-keton, cis-tujon, trans-tujon i kamfor su bili veoma zastupljeni, nagoveštavajući da bi možda upravo ove grupe jedinjenja mogli biti odgovorni za indukciju odbrane krompira.

Many aromatic, medicinal and ornamental species rich in biologically active secondary metabolites belong to the genus Tanacetum. Tanacetum vulgare L. (syn. Chrysanthemum vulgare L., tansy) is a perennial, herbaceous plant originating in Europe and Central Asia, commonly encountered along roads, stripes, pastures, fields, and coastal areas. In Serbia, this species is part of the ruderal flora. The research subject of this dissertation was the phytochemical characterization of the composition and biological activities of T. vulgare essential oil and methanol extracts. In vitro culture of T. vulgare was successfully established from seeds of plants harvested in nature. Shoot cultures have been successfully maintained and multiplied on the nutrient medium enriched with 6-benziaminopurine (BAP); for needs of rooting, BAP was replaced with indole buteric-acid (IBA). The root in vitro cultures were maintained in a liquid nutrient medium supplemented with IBA which influenced the increase in the biomass of cultivated roots during subcultures. Histological analysis of secretory structures of in vitro plantlets showed presence of biseriate glandular trichomes with detected lipids, terpenes and alkaloids as secretory content,, both on the leaves and the stalk. The chemical analysis of methanol extracts of T. vulgare herb and roots indicated that the extracts of in vitro cultivated plants were qualitatively poorer than methanol extracts of plants harvested in nature, but the extracts isolated from in vitro plants were characterized by several times higher presence of certain compounds, primarily 3,5-O-dicaffeoylquinic acid. The most common compounds were phenolic acids from the derivatives of cinnamic acid group, such as neohlorogenic, 3,5-O-dicaffeoylquinic,and dicaffeoylquinic acid. In addition, 17 flavonoids were detected. The relative proportion of phenolic acids and flavanoids differed depending on the part of the plant used to prepare the methanol extract. Comparative analysis of the chemical composition of the volatile components in essential oil of T. vulgare collected from the natural habitat and in vitro cultured plants has also shown great differences in the composition both of individual and entire groups of compounds. In the plants harvested in nature, GC/MS analysis showed that the dominant components belonged to the group of oxidized monoterpenes, with the most prevalent compounds of trans-chrysanthenyl acetate, trans-chryzanthenol, trans-thujone and cis- thujone. On the other hand, the essential oil obtained from in vitro grown T. vulgare plants was characterized by a uniform presence of monoterpenes and sesquiterpens. All monoterpenes identified in in vitro plantlets were also present in essential oil of plants collected from nature, on the other hand, sesquiterpene compounds were more diverse and more represent in in vitro plants. Essential oil and methanol extracts isolated from plants harvested from natural habitat were used to analyze the biological activities of the T. vulgare secondary metabolism compounds. The results showed that all the analyzed methanol extracts exhibited a significant antioxidant activity that correlated with the total phenol content in the extracts. The root extract showed the highest antioxidant capacity, containing thehighest amount oftotal phenols, with the maximum relative content of neochlorogenic, dicaffeoylquinic, and dicaffeoylquinic acids compared to other extracts. The essential oil and methanol extracts of T. vulgare flowers, leaves, stalks and roots exhibited significant antimicrobial activity tested on 8 bacterial and 6 micromycetes species. T. vulgare essential oil exhibited a strong antimicrobial effect on most bacterial strains (5 of the 8 analyzed) and all strains of micromycetes. On the other hand, methanol extracts of T. vulgare roots, leaves, flowers and stalks exhibited strong activity on all tested bacterial species and most of the micromycetes. In most cases the effects of the tested extracts were at the level of the effects of the reference antibiotics and antimycotics used as positive controls, so both essential oil and methanol extracts of T. vulgare can be recommended in the prevention and treatment of infections caused by species on which high activity is shown and also as preserving agents in the food industry. In further research, the cytotoxic potential of essential oil and methanol extracts on human cervical adenocarcinoma (HeLa) cells was investigated. To determine the level of selective effect, the activity on healthy human cell lines of fetal lung fibroblasts (MRC-5) was also studied. In vitro results showed that the essential oil exhibited the weakest cytotoxicity while methanol extracts of the stalk and root showed poor cytotoxic action on the target cells relative to the methanol extracts of leaf and flower. The most of HeLa cells exhibited loss of adhesion after leaf and flower methanol extracts treatment, also shrinking of membrane and rounding of cells were observed, indicating cell death. Monitoring the effects of T. vulgare essential oil on gypsy moth (Lymantria dispar L.) larvae showed that this essential oil did not exhibit acute toxicity to the caterpillar of the second developmental stage , but caused a delay in the coating and a significant reduction in the percentage of coated caterpillars. Ingestion of essential oil influenced on the other parameters of growth and development observed in the fourth stage of caterpillar development. The essential oil reduced the daily increment of the mass of caterpillars of the fourth developmental stage as well as the rate of food consumption. Although the essential oil influenced the reduction in the conversion of digested food into the caterpillar biomass, this reduction did not appear to be statistically significant. Hence, it can be concluded that the negative effect of essential oil on ingestion was reduced and that reduced intake of food was partially compensated. In order to investigate the potential of T. vulgare essential oil to induce the mechanisms of defense of in vitro grown potato plants (grown in an atmosphere filled with volatile components of this oil), the expression of 4 genes involved in the mechanisms of defense against pathogens and herbivore insects was analyzed. The presence of essential oil induced intense induction of two analyzed PR (pathogenesis- related) genes, PR-2 and PR-5. The first changes in expression were observed after 8 hours of exposure, and the maximum increase of expression of about 50 times was recorded for the PR-2 gene after 12 h. Based on the preliminary PTR-MS analysis and measurements of the representation of individual essential oil compounds in the atmosphere in jars where the potatoes were grown, the dominant compounds were the monoterpen hydrocarbons α-pinene and p- cymene. Beside these compounds, artemisia-ketone, cis-thujone, trans-thujone and camphor were highly represented, suggesting that these groups of compounds might be responsible for induction of potato defenses.

Nasleđivanje i ekspresija gena uključenih u otpornost suncokreta (Heliathus annuus L.) prema plamenjači

Inheritance and expression of genes involved in resistance of sunflower Helianthus annuus L.) to downy mildew

U radu je ispitan način nasleđivanja otpornosti prema patotipu plamenjače 730 kod tri populacije suncokreta. Inbred linija suncokreta Ha26S, koja je osetljiva na patotip plamenjače 730, je ukrštena sa linijama JM8, Ha338 i Ha26R koje su donori gena Pl za otpornost prema plamenjači. Nakon ukrštanja (Ha26SxJM8) proizvedene su 73 F3 familije koje su u ovom ispitivanju korišćene kao prva populacija na kojoj je ispitan način nasleđivanja otpornosti prema patotipu 730 plamenjače. Drugu populaciju čini 75 F3 familija koje su dobijene iz ukrštanja (Ha26SxHa338) a treću populaciju čine 73 F3 familije dobijene iz ukrštanja (Ha26SxHa26R). Otpornost linija i njihovog F3 potomstva na plamenjaču su analizirani na nivou: procene otpornosti koristeći tehniku inokulacije celog semena (primarna infekcija), kao i procene otpornosti na molekularnom nivou. Sve biljke na kojima nije došlo do sporulacije ocenjene su kao otporne. U prvoj populaciji, od svih ocenjenih F3 familija 52 familije su bile otporne, dok je 21 familija bila osetljiva. Rezultati testa otpornosti u drugoj populaciji pokazali su da je od svih ocenjenih F3 familija, 56 familija bilo otporno a 19 familija su osetljive prema patotipu 730 plamenjače. U trećoj populaciji 54 F3 familije ocenjene su kao otporne a 19 kao osetljive. Rezultati testa su potvrdili da odnos razdvajanja otpornih i osetljivih F3 familija odgovara odnosu 3 : 1 u sve tri ispitivane populacije. Na osnovu dobijenih rezultata potvrđeno je da je otpornost na patotip plamenjače 730 kontrolisana jednim genom. F3 familije su analizirane i na molekularnom nivou koristeći nekoliko PCR marker sistema. Način nasleđivanja, tj odnos razdvajanja za nekoliko markera za analoge gena otpornosti (Resistance Gene Analogues) i markera CAPS, u tri ispitivane populacije suncokreta testiran je χ2 testom. Testirana je hipoteza da je odnos RGA markera u nasleđivanju dominanto recesivan, odnosno da je odnos dobijenih fenotipova 3:1. Prema dobijenim rezultatima ova hipoteza je prihvatljiva za sve markere RGA osim za HaP1R i to samo u slučaju populacije u kojoj je korišćena JM8 linija suncokreta kao donor gena Pl6. U sve tri ispitivane populacije svi ostali markeri se uklapaju u dominatno – recesivi tip nasleđivanja, odnosno razdvajali su se u odnosu 3:1, bez obzira na roditeljske linije koje su bile izvori gena Pl. Markeri CAPS su se uvek razdvajali u odnosu 1:2:1, odnosno nasleđuju se kodominantno. Jedan od ciljeva ovog rada je bio da se utvrdi raspored dominantnih haplotipova RGA i kodominantnih CAPS markera na ,,linkejdžʼʼ mapama Pl6 regiona suncokreta u populacijama suncokreta koje su korišćene za mapiranje. Na ,,linkejdžʼʼ mapama dobijenim na tri ispitivane populacije za mapiranje dominantnih haplotipova RGA i kodominantnih CAPS markera, ispitivani markeri su raspoređeni na genetičkoj distanci od ~5 do ~12 cM. Uprkos razlikama koje postoje između mapa, raspored lokusa i "linkidž" razdaljine analiziranih markera su dobro održavane između mapa dobijenim za tri različite mapirajuće populacije. Na osnovu dobijenih rezultata može se zaključiti da kodominantni CAPS markeri za otpornost suncokreta prema patotipu plamenjače 730 mogu biti korišćeni u marker asistiranoj selekciji za ovu osobinu, bez obzira na roditeljske linije koje su korišćene kao izvori gena za otpornost. Međutim, rezultati dobijeni u ispitivanju populacije sastavljene od 37 RIL (Recombinant Inbred Lines) ukazuju na to da se u Pl6 genomskom regionu ipak javljaju promene čak i u kasnim generacijama samooplodnje u toku stvaranja biljaka otpornih na plamenjaču. U radu je pokazano da upotreba jednog dominantnog i kodominantnih CAPS markera ipak obezbeđuje efikasnu identifikaciju biljaka otpornih na patotip plamenjače 730, bez obzira na roditeljske linije koje se u početnom ukrštanju koriste kao izvori ili primaoci Pl gena. Jedan od ciljeva ovog rada je bio da se ispita ekspresija gena kandidata za otpornost ispitivanog u prvom delu, kao i gena Hacaox, Haoxox, HaSODc, HaSODp, HaSOD1, Hachi, HaPR5, HaEDS1, HaEDR1 koji su važni u odgovoru biljke na biotički stres prouzrokovan infekcijom gljivom Plasmopara halstedii. U listovima mladih biljaka dve linije suncokreta, od kojih je jedna otporna a druga osetljiva prema patotipu 730 plamenjače, praćena je ekspresija ovih gena, u periodu 2-96 sati nakon sekundarne infekcije sporama P. halstedii, odnosno nakon prskanja vodom u slučaju kontrolnih biljaka. Rezultati dobijeni sa Ha26R i Ha26S linijama suncokreta ukazuju da gen za otpornost koji je proučavan u ovom radu, ima konstitutivnu ekspresiju i u kontrolnom i u tretmanu inokulisanom suspenzijom spora P. halstedii. Nakon analize sa restrikcionim enzimima pokazano je da u otpornoj liniji suncokreta Ha26R, pored fragmenta koji je konstitutivno prisutan kod obe linije suncokreta (~800bp), postoji još jedan dodatni fragment koji je iste veličine, ali mu se sekvenca razlikuje u nekoliko SNP-ova (Single Nucleotide Polymorphism) u odnosu na konstitutivno prisutan fragment. Pretpostavka je da ovaj dodatni fragment koji je prisutan samo u otpornoj liniji Ha26R, obezbeđuje ovoj liniji otpornost na patotip 730 plamenjače. Analizom ekspresije gena Hacaox, Haoxox, HaSODc, HaSODp, HaSOD1, chi, HaPR5, HaEDS1, HaEDR1 ustanovljeno je da je većina ispitivanih gena imala konstitutivnu ekspresiju koja je izraženija u otpornoj liniji suncokreta. Ekspresija HaEDS1, Hachi, HaPR5, HaSODp, Hacaox i Haoxox kod otporne linije suncokreta je sukcesivno povećana u ranim fazama nakon infekcije. Najveća kvantitativna razlika u ekspresiji gena između otporne i osetljive linije suncokreta nakon infekcije suspenzijom spora P. halstedii, je indukcija ekspresije HaPR5 gena u ranim fazama nakon infekcije kod otporne linije. Dobijeni rezultati pokazuju da rani odgovor biljaka nakon sekundarne infekcije uzročnikom plamenjače, odgovara hipersenzitivnom odgovoru.

The inheritance of resistance to race 730 of sunflower downy mildew, in three mapping populations, was investigated in this thesis. Sunflower inbred line Ha26S, who is susceptible to sunflower downy mildew race 730 was crossed with lines JM8, Ha338 and Ha26R donors of Pl resistance genes for resistance to downy mildew. After crosses (Ha26SxJM8) 73 F3 families produced. These families were used as the first population for evaluation the inheritance of resistance to race 730 of sunflower downy mildew. The second populations represents 75 F3 families that were produced from the initial cross (Ha26SxHa338), and third population consists of 73 F3 families that were produced from the initial cross (Ha26SxHa26R). Resistance of lines and their F3 progeny to downy mildew were analyzed using the whole seedling inoculation technique (primary infection) as well as on a molecular level. All the plants who did not have symptoms of sporulation were evaluated as resistant. In the first population of all estimated F3 families 52 families were resistant, while 21 families were susceptible. In the second population results of the test of resistance have shown that of all the estimated F3 families, 56 families were resistant while 19 families are susceptible to downy mildew race 730. In the third population 54 F3 families were evaluated as resistant and 19 as susceptible. The results of the χ2 test confirmed that the segregation ratio of resistant to susceptible F3 families fit a 3:1 ratio in all examined mapping populations. The data confirm that the resistance to downy mildew race 730 is controlled by a single gene. The same material was studied on a molecular level by several PCR marker system. The inheritance of resistance i.e. the segregation of several RGA markers and CAPS markers in three mapping populations was tested by χ2 test. We tested the hypothesis that the ratio of RGA markers in inheritance is dominant / recessive, or that the obtained ratio of phenotypes fit a 3: 1. According to the results, this hypothesis is acceptable for all markers except for the HaP1R, only in the case of the mapping population who containing JM8 sunflower line as the donor of Pl6 gene. In all of three examined populations all other markers fit into the dominant - recessive type of inheritance, segregated in a 3:1 ratio, without regardless of the parental lines that were the sources of Pl genes. CAPS markers always segregated in a 1:2:1 ratio, i.e. have kodominant type of inheritance. One of the goals of this study was to determine the distribution of dominant haplotypes RGA and codominant CAPS markers on linkage maps of Pl6 genomic region in three sunflower mapping population. On linkage mapps determined in three sunflower mapping population, dominant haplotypes RGA and codominant CAPS markers are arranged on the genetic distance of ~ 5 to ~ 12 cM. Despite the differences that exist between maps, the loci order and linkage distances are well maintained between maps obtained for three different mapping population. Based on obtained results it can be concluded that codominant CAPS markers for the resistance of sunflower to downy mildew race 730 can be used in marker assisted selection for this trait, regardless of the parental lines used as sources of resistance genes. However, the results obtained in the sunflower population of 37 RIL (Recombinant Inbred Lines) indicate that changes in the genomic Pl6 region occur in the advanced generations in the course of breeding process. In this paper shown that the use of one dominant and codominant CAPS markers provide efficient identification of plants resistant to downy mildew race 730, regardless of the parental lines used as sources or acceptors of resistance genes. One of the objective of this study was to investigate the expression of Pl gene candidate for resistance, as well as genes Hacaox, Haoxox, HaSODc, HaSODp, HaSOD1, Hachi, HaPR5, HaEDS1, HaEDR1 that are important in plant response to biotic stress caused by Plasmopara halstedii infection. The expression of TIR-NBS- LRR gene in the leaves of young plants was examined in response to secondary downy mildew infection. Changes in gene expression, in response to secondary infection with suspension of spore of Plasmopara halstedii (pathotype 730), were investigated in two sunflower lines in the time interval 2-96h after infection, in order to follow the early response of these genes to the infection with this pathogen. In the young leaves of two sunflower lines, where one is susceptible and the other resistant to sunflower downy mildew race 730, the expression of these genes was examined in the range 2-96 hours after secondary infection with spores of P. halstedii, or treatment with wather. The results obtained on sunflower line Ha26S and Ha26R indicate that gene for resistance was expressed in both healthy and infected susceptible and resistant line. It was shown that after the treatment with the restriction enzymes, in the resistant sunflower line Ha26R, in addition to the fragment that is constitutively present in both sunflower lines (~ 800bp), there is the additional transcript of the same size differing in few SNPs. This additional fragment which is present only in the resistant line Ha26R probably provides resistance to pathotype 730 of sunflower downy mildew. The analysis of gene expression Hacaox, Haoxox, HaSODc, HaSODp, HaSOD1, chi, HaPR5, HaEDS1, HaEDR1 was shown that most examined genes were constitutive with higher expression in resistant line. The expression of genes HaEDS1, Hachi, HaPR5, HaSODp, Hacaox Haoxox were sequentially upregulated in the early stages after infection in resistant line. The highest quantitative differences between resistant and susceptible line upon infection was early induction of HaPR5 gene in resistant line. Obtained results indicate that the early response of defense related genes to secondary downy mildew infection, resembles to hypersensitive-like reaction.

Mogućnosti unapređenja rezilijentnosti urbane forme na klimatske promene

The Possibilities for improvement of urban form resilience to climate change

Uslovi neizvesnosti i dinamičnih promena koje je teško predvideti predstavljaju najveći izazov održivom razvoju savremenih gradova. Jedan od procesa koji u velikoj meri doprinose ovim uslovima su klimatske promene, za koje postoje jaki dokazi da su izazvane antropogenim uticajima, pre svega upotrebom fosilnih goriva i emisijom gasova izazivača efekta staklene bašte. Gradovi, u kojima danas živi većina svetskog stanovništva, mesta su najvećih uzročnika klimatskih promena, ali ujedno i njenih najizraženijih posledica. U centru pažnje ove disertacije je rezilijentnost gradova, odnosno sposobnost da se izbore sa nepredvidivim uticajima poput klimatskih promena, zadržavajući pritom osnovne činioce sopstvenog identiteta, strukture i procesa, kao i održivost svog razvoja. Oni se u ovom istraživanju posmatraju kao kompleksni socio-ekološki sistemi, koji se nalaze u stanju dis-ekvilibrijuma i koji se odlikuju neodređenošću, samo-organizacijom i prostornom ekstenzijom u kojima se sistemi funkcionišu po principu „odozgo na gore“. U jedinstvenom okviru „ekologije grada“ urbane forme se, kao proizvod ljudske aktivnosti adaptacije prirodnog okruženja prema sopstvenim potrebama, razmatraju zajedno sa ostalim fizičkim komponentama sistema, a društveni procesi kao deo ukupnih bioloških aktivnosti. Oslanjajući se na saznanja o uticajima koji različitih urbanih obrasci mogu imati po procese i performanse urbanog sistema, bavićemo se njihovom ulogom i značajem za opštu rezilijentnost urbanih sistema, kao i na onu posebnu koja se odnosi na klimatske promene. Urbane forme, kao „proizvod“ određenih modela urbanog dizajna, reflektuju specifične vrednosti društveno-ekonomskog sistema i njegovog odnosa prema životnoj sredini. Ovi modeli uspostavljaju raznovrsne i međusobno različite relacije između artificijelnih i prirodnih komponenti sistema, dajući bitno drugačiju sliku i karakteristike izgrađenog predela. Osnovni cilj istraživanja je utvrditi uticaj određenog modela urbanog dizajna (stoga i rezultujućih urbanih formi) na rezilijentnost socio-ekološkog sistema. Na osnovu urbomorfoloških, ekoloških i socioloških kriterijuma, osnovni zadatak je evaluirati rezilijentnost određenih obrazaca urbanih formi na klimatske promene, a zatim razmotriti pristupe urbanog dizajna u cilju njihovog unapređenja.

The uncertain conditionas and unpredictable dynamic changes are the biggest challenge for sustainable development of contemporary cities. One of the processes that have major contribution to these conditions is climate change, a process that strong evidence imply is caused by anthropogenic factors, primarily by the use of fossil fuels and emissions of greenhouse gases. Cities, which contain the majority of today’s world population are the places of the biggest causes of climate change, but also the places of its most pronounced effects. The main focus of this thesis is the resilience of cities, e.g. the ability of city to cope with unpredictable impacts such as climate change, while maintaining the fundamental properties of its own identity, structures and processes, as well as the sustainability of its development. They are observed in this study as a complex socio-ecological systems, which are in the state of dis- equilibrium, and characterized by uncertainty, self-organization and spatial extension in which systems operate based on the "top-down" principle. In the integral framework of the "ecology of city" the urban form, as a product of human activities of adaptation natural environment according to their needs, will be considered together with other physical components of the system, and social processes as a part of the overall biological activity. Based on the findings of the impacts that different urban patterns could have on the processes and performance of urban systems we will deal with their role and importance to the overall resilience of urban systems as well as a specific one that refers to climate change. Urban form, as a "product" of certain model of urban design, reflects specific values of certain socio- economic system, as well as its relationship to the environment. These models have very diverse and disparate relations between artificial and natural components of the system, giving a rather different picture and the characteristics of the built landscape. The main objective of the thesis is to determine the influence of a particular model of urban design (hence the resulting urban form) to the resilience of socio-ecological system. Based on morphological, environmental and social criteria, the main objective is to evaluate resilience of certain patterns of urban form to climate change, and then consider urban design approaches that could improve it.

Taksonomsko-morfološka i molekularna studija troglobiontnih trčuljaka (Coleoptera: Carabidae: Trechinae) Karpato-balkanida Srbije

Taxonomic, morphological and molecular study of the troglobitic ground beetles (Coleoptera: Carabidae: Trechinae) of the Carpatho-Balkanids of Serbia

Tribus Trechini (potporodica Trechinae) je jedna od glavnih grupa insekata čiji su pripadnici kolonizovali podzemnu sredinu. Veliki broj taksona ovih insekata je razvio slične morfološke adaptacije na život u podzemnim staništima, što znatno otežava utvrđivanje njihovih filogenetskih odnosa. Ciljevi ove studije su fokusirani na rasvetljavanje taksonomsko-filogenetskih odnosa trehina koji nastanjuju pećine i jame Karpato-balkanida Srbije pomoću taksonomsko-morfoloških i molekularnih analiza. Taksonomsko-morfološke analize su obuhvatile prikaze morfoloških karakteristika studiranih taksona, dobijene pomoću klasične svetlosne i nelinearne mikroskopije (TPEF modalitet), kao i komparativne analize navedenih taksona. Molekularne analize su bile zasnovane na parcijalnim sekvencama gena za podjedinicu I citohrom c oksidaze (COI). Klasičnom svetlosnom mikroskopijom ispitane su morfološke karakteristike habitusa i genitalnih struktura svih analiziranih taksona, a analiza genitalnih i drugih anatomskih struktura predstavnika roda Pheggomisetes urađena je dodatno pomoću nelinearne mikroskopije (TPEF modalitet). Molekularnim analizama su podvrgnuti svi taksoni iz ispitivane oblasti, a uspešno je sekvencionirana DNK sledećih taksona: Pheggomisetes globiceps ninae, P. globiceps globiceps, P. globiceps ciniglavcensis, P. globiceps ilandjievi, P. serbicus serbicus, P. serbicus belensis, Glabroduvalius tupiznicensis, Curcicia bolei, Duvalius (Paraduvalius) sp., D. (P.) trifunovici, D. (P.) rtanjensis, D. (P.) milutini, D. (P.) stankovitchi georgevitchi i D. (P.) stankovitchi devojensis. Nakon sprovedenih analiza, dat je pregled taksona opisanih od strane drugih autora: D. (P.) stankovitchi [sa podvrstama D. (P.) stankovitchi stankovitchi, D. (P.) stankovitchi devojensis i D. (P.) stankovitchi georgevitchi], D. (P.) winkleri, D. (P.) rtanjensis provalijae i Curcicia (sa vrstom C. bolei). Takođe je dat pregled taksona opisanih od strane autora: D. (P.) trifunovici, D. (P.) rtanjensis, D. (P.) bogovinae, D. (P.) milutini, D. (P.) beljanicae, D. (P.) petrovici, D. (P.) sotirovi, P. globiceps ciniglavcensis, P. serbicus (sa podvrstama P. serbicus serbicus i P. serbicus belensis) i Glabroduvalius (sa vrstom G. tupiznicensis). Pored toga, potvrđen je status roda Curcicia i promenjen je status taksona P. ninae, prvobitno opisanog kao nezavisna vrsta, u podvrstu u okviru P. globiceps – P. globiceps ninae.

Tribe Trechini (subfamily Trechinae) is one of the main groups of insects whose members colonized underground environment. Large number of the insect taxa developed similar morphological adaptations due to life in underground habitats, which makes it difficult to determine their phylogenetic relationships. The aims of this study are focused on revealing taxonomic and phylogenetic relationships within Trechini inhabiting caves and pits of the Carpatho-Balkanids of Serbia, using taxonomic, morphological and molecular analyses. Taxonomic and morphological analyses included the displays of the morphological characteristics of the studied taxa obtained by classical bright-field and nonlinear microscopy (TPEF modality), as well as comparative analyses of the mentioned taxa. Molecular analyses were based on the barcoding region of cytochrome c oxidase subunit I (COI) gene. Morphological characteristics of habitus and genital structures of all analyzed taxa were examined by classical bright-field microscopy, and the analyses of genital and other anatomical structures of Pheggomisetes taxa were additionally examined by nonlinear microscopy (TPEF modality). All underground trechine taxa from the investigated region were subjected to molecular analyses, and we sucessfully isolated DNA of the following taxa: Pheggomisetes globiceps ninae, P. globiceps globiceps, P. globiceps ciniglavcensis, P. globiceps ilandjievi, P. serbicus serbicus, P. serbicus belensis, Glabroduvalius tupiznicensis, Curcicia bolei, Duvalius (Paraduvalius) trifunovici, D. (P.) rtanjensis, D. (P.) milutini, D. (P.) sp., D. (P.) stankovitchi georgevitchi and D. (P.) stankovitchi devojensis. After performed analyses, the review of the taxa described by other authors is given: D. (P.) stankovitchi [with subspecies D. (P.) stankovitchi stankovitchi, D. (P.) stankovitchi devojensis and D. (P.) stankovitchi georgevitchi], D. (P.) winkleri, D. (P.) rtanjensis provalijae and Curcicia (with species C. bolei). Also, the review of the taxa described by the author is given: D. (P.) trifunovici, D. (P.) rtanjensis, D. (P.) bogovinae, D. (P.) milutini, D. (P.) beljanicae, D. (P.) petrovici, D. (P.) sotirovi, P. globiceps ciniglavcensis, P. serbicus (with subspecies P. serbicus serbicus and P. serbicus belensis) and Glabroduvalius (with species G. tupiznicensis). Furthermore, the status of the genus Curcicia is confirmed and the status of the taxon P. ninae, previously described as a separate species, is changed to P. globiceps ninae, a subspecies within species P. globiceps.

Imunomodulatorna aktivnost rekombinanatne izoforme lektina iz banane u fiziološkim i patološkim uslovima u debelom crevu miševa BALB/c soja

Immunomodulatory activity of recombinant banana lectin isoform in large intestine of BALB/c mice under physiological and pathological conditions

Lektin banane (BanLec) pripada podgrupi lektina koji vezuju glukozu ili manoza- u okviru familije lektina sličnih žakalinu. U plodu banane, koji predstavlja prirodni izvor BanLec-a, javlja se u više izoformi, a može se proizvesti i rekombinantnom DNK tehnologijom. Rekombinantna izoforma BanLec-a (rBanLec) je po svojim strukturnim karakteristikama i specifičnosti vrlo slična prirodnim izoformama. Kao i većina biljnih lektina, (r)BanLec ne podleže brzo denaturaciji / razgradnji u uslovima digestivnog trakta, a pokazano je da se vezuje za epitel tankog creva i postepeno prolazi u subepitelni prostor. Danas je poznato da BanLec, nezavisno od izoforme, ima imunomodulatorni potencijal. Dosadašnja istraživanja su se dominantno bavila uticajem (r)BanLec-a na funkcionalne karakteristike limfocita i pokazano je da stimulacije određene ćelijske populacije prirodnim i rekombinantnom izoformom kvalitativno imaju isti ishod. Uticaj (r)BanLec-a na funkcionalne karakteristike drugih ćelija imunskog sistema nije detaljno analiziran. Cilj ove doktorske teze je da se kroz ispitivanja (1) modulatornog dejstva rBanLec- a na funkcionalne karakteristike antigen-prezentujućih ćelija (APĆ) BALB/c miša, (2) imunomodulatornog dejstva rBanLec-a u debelom crevu BALB/c miša u fiziološkim uslovima, i (3) efekata profilaktičke i terapijske primene rBanLec-a u modelu TNBS-om indukovanog kolitisa kod BALB/c miša, dobije uvid u imunomodulatorni efekat rBanLec-a na imunski sistem u mukozi debelog creva miševa BALB/c soja pod specifičnim uslovima. Koristeći peritonealne makrofage i dendritske ćelije slezine (DĆ) kao in vitro model sisteme, pokazano je da rBanLec dozno-zavisno moduliše funkcionalne karakteristike APĆ BALB/c miševa. Analiza uticaja rBanLec stimulacije na karakteristike rezidentnih (RM) i tioglikolatom-indukovanh (TGM) peritonealnh makrofaga, pokazala je da ishod stimulacije određene ćelijske populacije nije jednoznačan već zavisi od njihovog funkcionalnog stanja. rBanLec na pozitivan dozno-zavisan način pospešuje proinflamatorni kapacitet RM (povećanje produkcije NO i IL-12 i smanjenje aktivnosti arginaze i sekrecije IL-4), a na isti način smanjuje proinflamatorni kapacitet TGM (smanjenje produkcije NO i IL-12 i povećanje aktivnosti arginaze i sekrecije IL-4). Uticaj rBanLec-a na produkciju TGF-β kod peritonealnih makrofaga takođe zavisi od funkcionalnog stanja makrofaga (kod RM, rBanLec negativno dozno-zavisan, a kod TGM pozitivno dozno-zavisno utiče na produkciju TGF-β). I kod RM i kod TGM, rBanLec na pozitivan dozno-zavisan način podstiče aktivnost mijeloperoksidaze (MPO), produkciju TNF-α i IL-10. Za stimulaciju produkcije proinflamatornih medijatora, nezavisno od funkcionalnog stanja makrofaga, značajno je vezivanje rBanLec-a za TLR2 i TLR4 / CD14. rBanLec utiče na sekreciju efektorskih citokina IFN-γ i IL-4 (negativno dozno-zavisno) i regulatornog citokina IL-10 (pozitivno dozno-zavisno) od strane DĆ slezine. rBanLec TLR2-posredovana stimulacija DĆ slezine nije ključna za promene u ekspresiji Ifn-γ i Il-4. U okviru ove doktorske teze je pokazano da rBanLec stupa u interakciju sa ćelijama epitela debelog creva i subepitelnog odeljka, i moduliše lokalne imunske karakteristike. Imunomodulatorni efekat rBanLec-a u debelom crevu je dozno-zavisan. Jednokratno lokalno aplikovan rBanLec u dozama od 0,5 do 50 µg/kg TT u debelom crevu BALB/c miša dozno-zavisno stimuliše mehanizme urođene imunosti koji deluju antimikrobno (MPO i iNOS aktivnost), pri čemu njihova pojačana aktivnost ne dovodi do lokalnih patoloških promena. rBanLec u dozi od 0,5 µg/kg TT (0,1 µg/ml) ne pokreće produkciju proinflamatornih citokina ali stimuliše produkciju regulatornih citokina, dok u dozama > 5 µg/kg TT (>1 µg/ml) stimuliše uspostavljanje balansiranog proinflamatornog imunskog odgovora Th1 / Th17 tipa. Već tokom prvih 24h, rBanLec, u koncentracijama 0,1-10 µg/ml, indukuje značajan porast lokalne koncentracije IL-2 čija koncentracija pozitivno korelira sa aplikovanom dozom rBanLec-a. Do značajnog porasta lokalne koncentracije IFN-γ, IL-12 i IL-17 dolazi tek drugog dana i ovaj porast je prolaznog karaktera. Takođe, drugog dana, nezavisno od lokalno aplikovane količine rBanLec-a, dolazi i do značajnog porasta lokalne koncentracije IL-10. U modelu TNBS-om indukovanog kolitisa kod BALB/c miševa pokazano je da rBanLec, u dozama 0,5-50 µg/kg TT, može da ima profilaktički i pozitivan terapijski efekat u slučaju inflamatorne bolesti creva. Jednokratno lokalno primenjen rBanLec u navedenim dozam ne može da spreči razvoj eksperimentalnog kolitisa, ali su određeni profilaktički tretmani rezultovali blažom kliničkom slikom u piku bolesti i bržim oporavkom miševa. Stepen poboljšanja opšte kliničke slike zavisio je i od profilaktičke doze rBanLec-a i od vremena njene primene. Terapijska primena rBanLec-a koja podrazumeva svakodnevnu oralnu primenu rBanLec-a u dozama 0,5-50 µg/kg TT počevši od dana indukcije eksperimentalnog kolitisa, takođe, na dozno-zavisan način, ublažavala opštu kliničku sliku. U slučaju profilaktičke i u slučaju terapijske primene rBanLec-a, opšte poboljšanje kliničke slike rezultat je ne samo smanjenja intenziteta lokalnog zapaljenskog procesa (smanjenje: aktivnosti MPO i iNOS, ekspresije transkripcionih faktora T-bet i RORγt i koncentracije IL-12 i TNF-α), nego i promene odnosa aktivnosti proinflamatornih i regulatornih mehanizama. Stepen poboljšanja kliničke slike u piku bolesti usled specifičnog rBanLec tretmana je pozitivno korelirao sa smanjenjem odnosa [TNF-α] / [IL-10] u debelom crevu. Rezultati prikazani u ovoj doktorskoj disertaciji pokazuju da rBanLec, modulišući aktivnost ćelija imunskog sistema, utiče na karakteristike lokalnog miljea u debelom crevu. Lokalno imunomodulatorno dejstvo rBanLec-a može doprineti ublažavanju opšte kliničke slike u slučaju inflamatorne bolesti creva.

Banana lectin (BanLec) is primarily isolated from the fruit of banana (Musa paradisiac). It is glucose / mannose-specific lectin that belongs to the jackalin superfamily of lectins. There are several naturally occurring BanLec isoforms. Nowadays BanLec can also be produced by recombinant technology (rBanLec). rBanLec possesses structural and functional characteristics that highly resemble those reported for its natural counterparts. As most plant lectins, (r)BanLec is resistant to denaturation/proteolytic cleavage in the gastrointestinal tract. It has been reported that rBanLec attaches specifically to the mucosal surface of small intestine and passes into subepithelial compartment. (r)BanLec has been recognized as a potential immunomodulator. It has been shown that (r)BanLec modulates functional characteristics of lymphocytes but the effects of (r)BanLec stimulation in other immune cells are not yet elucidated. The aim of this study was to investigate immunomodulatory / immunostimulatory activity of rBanLec in the colon of BALB/c mice. This aim was accomplished through evaluation of the impact of rBanLec stimulation on 1) functional characteristics of antigen presenting cells (APC) isolated from BALB/c mice 2) local immune system in the large intestine of healthy BALB/c mice, and 3) the course of TNBS-induced experimental colitis in BALB/c mice. Peritoneal macrophages and spleen dendritic cells (DC) isolated from BALB/c mice were used in vitro for the evaluation of rBanLec influence on functional characteristics of APC. Generally, it is shown that rBanLec in a dose-dependent manner modulated the functional characteristics of APCs. By using resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages, it has been shown that effects of rBanLec stimulation depend on its concentration but also on the functional status of macrophages. Obtained results have clearly shown that rBanLec, in a positive dose- dependent manner, promotes pro-inflammatory phenotype with BALB/c RMs (enhancement of NO and IL-12, reduction of arginase activity and IL-4 secretion) while, in the same manner, it tends to skew BALB/c TGMs towards anti-inflammatory profile (reduction of NO and IL-12 production, enhancement of arginase activity and IL-4 secretion). The dose-dependent changes in production of TGF-β by peritoneal macrophages also depended on their functional status: rBanLec stimulated the production of TGF-β by RMs in a negative dose-dependent manner, while in TGMs production of TGF-β positively correlated to the rBanLec concentration. The activity of myeloperoxidase (MPO) and productions of TNF-α and IL-10 were enhanced upon rBanLec stimulation in a positive dose-dependent manner with both RMs and TGMs. Further, it was shown that interactions of rBanLec with TLR2 and TLR4 / CD14 are important for initiations of the production of pro-inflammatory mediators by peritoneal macrophages irrespective to their functional status. rBanLec also stimulated in a specific dose-dependent manner the secretion of effector cytokines IFN-γ and IL-4 (negative dose- dependent manner) and regulatory cytokine IL-10 (positive dose-dependent manner) by spleen DCs. However, it seems that TLR2-mediated rBanLec stimulation is not crucial for changes in the expression of Ifn-γ and Il-4 in spleen DCs. It has been shown that rBanLec binds to the luminal surface of murine colon epithelium, passes through epithelial barrier and modulates the local immune response. Single rectal application of rBanLec in doses ranging from 0.5 to 50 μg/kg BW (0.1 - 10 μg/ml) triggered the mechanisms of innate immunity (MPO and iNOS activity), which contribute to the antimicrobial activity in the colon of BALB/c mice. Although the enhanced activity of MPO and iNOS was recorded upon the rBanLec stimulation, no pathology changes in the colon tissue were marked. rBanLec in a dose of 0.5 μg/kg BW (0.1 μg/ml) did not stimulate the production of pro-inflammatory cytokines but enhanced the production of regulatory cytokines. In doses ≥ 5 μg/kg BW (≥ 1 μg/ml), rBanLec promoted the transitional establishment of balanced Th1/Th17 local immune response. A significant increase in local concentration of IL-2 that positively correlated with the applied rBanLec dose was marked during the first 24 hours upon stimulation with rBanLec in doses ranging from 0.1 to 10 μg/ml. The transitional rise in local production of IFN-γ, IL-12 and IL-17 was marked only upon treatment with rBanLec in concentration ≥ 1 μg/ml and it occurred during the second day upon rBanLec administration. In addition, irrespective of the applied dose of rBanLec, a significant increase of the local concentration of IL-10 was recorded on the second day upon rBanLec administration. rBanLec, within a dose range analyzed herein (0.5-50 μg/kg BW) might exert prophylactic and positive therapeutic effects on inflammatory bowel disease. Prophylactic application of rBanLec in specified doses did not prevent the development of TNBS- induced colitis in BALB/c mice, but some treatments resulted in less severe pathology in the peak of disease and improved the recovery process. The extent of positive effect of prophylactic treatment with rBanLec depended on both rBanLec dose and the time of its application. Therapeutic treatment, which assumed oral administration of rBanLec in doses from 0.5 to 50 μg/kg BW on daily basis starting from the day of the induction of experimental colitis, also ameliorated disease severity in a dose-dependent manner. Reduction in experimental colitis severity upon prophylactic or therapeutic rBanLec treatment was due to lessening of the intensity of local inflammatory processes (local reductions in: activity of MPO and iNOS, expression of transcription factors T-bet and RORγt, and concentrations of IL-12 and TNF-α), but also due to the stimulation of local regulatory mechanisms. The extent of reduction in disease severity due to specific rBanLec treatment positively correlated to the decrease in the [TNF-α] / [IL-10] ratio in the colon. The results presented in this thesis showed that rBanLec modulates the activity of the immune cells in a specific dosage-dependent manner and affects the characteristics of the local milieu in the murine colon. Local immunomodulatory impact of rBanLec may contribute to the alleviation of symptoms in the inflammatory bowel diseases.

Molekularno-genetička osnova kongenitalne adrenalne hiperplazije u Srbiji: karakterizacija mutacija u CYP21A2 genu

Molecular-genetic basis of congenital adrenal hyperplasia in Serbia: characterization of mutations in CYP21A2 gene

Kongenitalna adrenalna hiperplazija (KAH) obuhvata grupu autozomno recesivnih oboljenja okarakterisanih narušenom sintezom steroidnih hormona u kori nadbubreţne ţlezde i širokim spektrom kliniĉkih simptoma. Na osnovu fenotipske ekspresije razlikuju se klasiĉna (jednostavna virilizujuća i forma sa gubitkom soli) i neklasiĉna forma bolesti. U više od 90% sluĉajeva javlja se zbog deficijencije enzima steroid 21-hidroksilaze (CYP21A2) kao posledice prisustva mutacija u CYP21A2 genu. Dva osnovna mehanizma dovode do inaktivacije CYP21A2 gena: nejednaka razmena delova homologih sekvenci izmeĊu nesestrinskih hromatida u mejozi moţe dovesti do formiranja CYP21A1P/CYP21A2 himernih gena ili delecije celog CYP21A2 gena, dok se putem genske konverzije u aktivan gen kopiraju homologe, ali defektne sekvence CYP21A1P pseudogena. Zbog varijabilnosti i kompleksne strukture regiona u kome je lociran CYP21A2 gen molekularna dijagnostika KAH-a i danas predstavlja izazov. Ovo je prva studija molekularno-genetiĉke osnove KAH-a u Srbiji. Izvršena je optimizacija PCR-SSP i metode direktnog DNK sekvenciranja CYP21A2 gena ĉime je omogućena detekcija CYP21A1P/CYP21A2 himernih gena, mutacija nastalih genskom konverzijom, ali i retkih i novih genetiĉkih varijanti. UtvrĊen je celokupan spektar mutacija kao i njihove uĉestalosti kod 61 nesrodnog pacijenta. Identifikovano je 18 alela sa patogenim efektom i dostignut je nivo detekcije mutacija od 82,0%. Nivo detekcije je bio najviši (94,7%) u grupi pacijenata sa najteţom formom bolesti (klasiĉni KAH sa gubitkom soli). Najuĉestalija mutacija u populaciji iz Srbije, kao i u većini drugih populacija, je c.290-13A/C>G (18,5%). Pet uĉestalih alela koji ukljuĉuju c.290-13A/C>G, CYP21A1P/CYP21A2, p.P30L, p.R356W i p.G110fs mutacije ĉine 63,0% svih mutiranih CYP21A2 alela u analiziranoj grupi pacijenata. Primećena je neuobiĉajeno visoka uĉestalost p.R356W mutacije (11,1%), kao i kompleksnih alela sa višestrukim mutacijama (7,4%), meĊu kojima ĉak 75,0% sadrţi p.P453S mutaciju...

Congenital adrenal hyperplasia (CAH) comprises of group of autosomal recessive disorders characterized by impaired adrenal steroidogenesis and wide spectrum of clinical manifestations. There are two main phenotypes: classical (salt wasting and simple virilizing) and non-classical form. In more than 90% of patients the disease is caused by steroid 21-hydroxylase (CYP21A2) deficiency resulting from underlying CYP21A2 gene mutations. Two major molecular mechanisms lead to CYP21A2 gene inactivation: unequal crossing over results in CYP21A1P/CYP21A2 chimerae formation or whole CYP21A2 gene deletion, whereas gene conversion copy homologue, but defective sequences from CYP21A1P pseudogene to the active gene. Due to variability and complex structure of the region where CYP21A2 gene resides molecular diagnostics of congenital adrenal hyperplasia remains a challenge. This is a first comprehensive study regarding molecular genetic basis of CAH in Serbia. Optimization of PCR-SSP and direct DNA sequencing of CYP21A2 gene was conducted, thus enabling detection of CYP21A1P/CYP21A2 chimerae, pseudogene derived mutations, but also rare and new genetic variants. Analysing 61 unrelated patients complete mutational spectrum, as well as mutation’s frequencies, was determined. Eighteen pathogenic alleles were identified and detection rate of 82.0% was assessed. Detection rate was highest (94.7%) in patients with severe classical form with salt wasting. The most frequently occurring mutation in patients from Serbia, as well as in many other populations, is c.290-13A/C>G (18.5%). Five alleles including c.290-13A/C>G, CYP21A1P/CYP21A2, p.P30L, p.R356W and p.G110fs mutations account for 63.0% of all mutated CYP21A2 alleles in analysed patients. Unusually, high frequency of p.R356W mutation (11.1%), as well as complex alleles with multiple mutations (7.4%), most frequently including p.P453S (75.0%), was detected. Direct DNA sequencing unravelled two new missense mutations p.L129P and p.S165P, for which computer predictions showed to be probably pathogenic...

Microbiological quality of water and detection of genotoxic pollution in different sectors of the Sava river with prokaryotic and eukaryotic test systems

Микробиолошки квалитет воде и детекција генотоксичног загађења различитих сектора реке Саве прокариотским и еукариотским тест системима

Sliv reke Save je pod snažnim uticajem stresora antropogenog porekla, međutim posledice koje stresori mogu imati na vodenu biotu nisu dovoljno ispitane. Ova disertacija ima za cilj da ispita genotoksični potencijal duž reke Save. Istraživanje je organizovano u dve celine. U oviru prve celine ispitivalo se koji bioindikatorski organizam je optimalan za istraživanje na nivou čitave reke. Takođe ispitivao se uticaj sezone na odgovor biomarkera kao i metodologija očuvanje uzorka na rezultate analiza. U okviru druge celine rađeno je kompleksno istraživanja duž 900 rkm reke Save. Korišćen je kombinovani in vivo i in vitro pristup, pri čemu je analizirano ukupno 12 lokacija. Genotoksični potencijal je praćen upotrebom složene baterije biotestova na prokariotskim i eukariotskim organizimima. Baterija je obuhvatala procenu mutagenog potencijala SOS/umuC testom na bakteriji Salmonella typhimurium TA1535/pSK1002. Nivo oštećenja DNK, kao biomarker izlaganja (kometni test) i biomaker efekta (mikronukleus test) kao i nivo oksidativnog stresa (Fpg - modifikovani kometni test) praćen je u krvnim ćelijama riba (Alburnus alburnus/Alburnoides bipunctatus). Rezultati su pokazali različitu osetljivost primenjenih bioloških testova u detekciji genotoksičnog potencijala. Komet test je imao veći potencijal u diferenciranju lokaliteta po genotoksičnom potencijalu u poređenju sa drugim primenjenim testovima. Podaci predstavljeni u ovoj disertaciji predstavljaju sliku trenutnog stanja reke. Genotoksični potencijal zabeležen duž reke se može vezati za narušen kvalitet vode uticajem komunalnih i industrijskih otpadnih voda. Podaci prikazani u ovoj studiji potiču iz jedinstvenog izvora (homogenost analiza za sve uzorke) uprkos velikoj geografskoj površini koju studija pokriva što joj dodatno daje na značaju.

The Sava River Basin is under strong impact of stressors related to anthropogenic activity, yet the consequences that the stressors might have on aquatic biota are not properly addressed. This dissertation aims to provide data on genotoxic potential along the Sava River. Research was organized in two campaigns. In the first campaign, preliminary research was conducted to indicate optimal bioindicator, season, sample preservation and bioassays for performing the research on the scale of the whole Sava River. The second campaign consists of comprehensive survey along the 900 rkm of the Sava River. With combined in vivo and in vitro approach, 12 sites were analyzed in total. The genotoxic potential was assessed using a complex battery of bioassays performed in prokaryotes and aquatic eukaryotes (freshwater fish). Battery comprised evaluation of mutagenicity by SOS/umuC test in Salmonella typhimurium TA1535/pSK1002. The level of DNA damage as a biomarker of exposure (comet assay) and biomarker of effect (micronucleus assay) and the level of oxidative stress as well (Fpg - modified comet assay) was studied in blood cells of bleak and spirlin (Alburnus alburnus/Alburnoides bipunctatus respectively). Results indicated differential sensitivity of applied bioassays in detection of genotoxic pressure. Comet assay showed higher potential in differentiation of the sites based on genotoxic potential in comparison with other assays. Data presented in this dissertation are a snapshot of the current status of the river. Genotoxic potential along the river can be traced to the deterioration of water quality by communal and industrial wastewaters. The major highlight of the study is that the complex set of data was obtained from a single source (homogeneity of analyses for all samples).

Medonosna pčela (Apis mellifera) kao bioindikator zagađenja toksičnim metalima na teritoriji Srbije

Honeybees (Apis mellifera) as bioindicators of toxic metal pollution in Serbia.

Povećanje broja stanovnika na Zemlji neminovno dovodi i do povećanja antropogenog zagađenja. Metali se smatraju jednim od najopasnijih zagađivača današnjice, posebno zbog toga što mnogi od njih ne podležu biodegradaciji. Uzimajući ovo u obzir, neophodno je pronaći metode za njihovo praćenje u životnoj sredini koje su jeftinije, pokrivaju veća područja i daju brže rezultate. Jedna od alternativa klasičnim tehnikama merenja jeste upotreba bioindikatorskih organizama. U ovom radu korišćena je medonosna pčela (Apis mellifera L.) kao bioindikator zagađenja toksičnim metalima na teritoriji Srbije. U telima medonosnih pčela sakupljenih iz pčelinjaka u Beogradu, Kostolcu i Mesiću tokom dva meseca (jul i septembar) 2014. godine analizirani su Al, Ba, Cd, Co, Cr, Cu, Fe, Li, Mn, Na, Ni, Pb, Sr i Zn. U telima pčela uzorkovanih u Pančevu i Vršcu tokom septembra 2013. godine, jula i septembra 2014. i jula i septembra 2015. godine analizirani su Al, Ba, Cd, Co, Cr, Cu, Fe, Mn, Na, Ni, Sr i Zn. Pčele su uzorkovane u ovom periodu jer je njihova aktivnost usko vezana za vremenske prilike, a ova dva meseca predstavljaju vrhunac te aktivnosti. Isti metali su analizirani i u uzorcima zemljišta uzetih u blizini pčelinjaka u Pančevu i Vršcu. Za određivanje koncentracije ovih metala korišćena je optička emisiona spektrometrija sa induktivno spregnutom plazmom (ICP-OES). Dvosmerna ANOVA je pokazala da postoje statistički značajne razlike između koncentracija pojedinih metala u Beogradu, Kostolcu i Mesiću. Značajne razlike između bar dve lokacije ustanovljene su za Al, Ba, Cr, Cu, Fe, Li i Ni. Korišćenjem analize glavnih komponenti (PCA), klasterske analize (CA) i ANOVA-e, ustanovljeno je da povećane koncentracije Al, Fe i Cr u okolini Kostolca potiču iz termoelektrana „Kostolac A i B“ i vi pripadajućeg pepelišta. Visoke koncentracije Cu na teritoriji Beograda pripisane su saobraćaju i upotrebi kočnica koje sadrže Cu. Visoke koncentracije Ni u Mesiću potiču od upotrebe organskog i veštačkog đubriva, dok više koncentracije Ba i Cu na istoj lokaciji potiču od sredstava za prskanje voća i vinograda koji se nalazi u neposrednoj blizini Mesića. Poređenjem koncentracija metala između Vršca i Pančeva, došlo se do zaključka da su koncentracije Al, Ba i Sr statistički značajno više u Vršcu. Pomoću PCA, CA i korelacija između metala u zemljištu i onih između metala u pčelama, došlo se do zaključka da Co, Cd, Na, Fe, Mn i Zn detektovani na teritoriji Vršca i Pančeva potiču iz antropogenih izvora, prevashodno od „Hesteel Serbia Iron & Steel“ (nekadašnje „Železare Smederevo“), termoelektrane „Kostolac A i B“, saobraćaja, kao i intenzivne poljoprivrede. Ca, Al, Mg i Cr sa istih lokacija imaju prirodno poreklo, dok neki elementi (Cu, Ba, Sr, Ni) potiču kako iz antropogenih tako i iz prirodnih izvora. Tokom 2014. godine analizirani su metali u pčelama sa svih pet lokacija: Beograd, Pančevo, Kostolac, Vršac i Mesić. Upotrebom ANOVA-e, PCA i CA ustanovljeno je da su koncentracije Ba i Cd više u Mesiću u poređenju sa drugim lokacijama, što se može generalno pripisati intenzivnoj poljoprivredi u tom kraju. Više koncentracije Fe i Cr u Kostolcu potiču od termoelektrane „Kostolac A i B“, dok su značajno više koncentracije Cu u Beogradu posledica intenzivnog saobraćaja. Sezonske varijacije u koncentracijama metala uočene su za Al, Ba, Ca, Cd, Co, Cu, Fe, Mg i Sr. U većini slučajeva primećeno je da su koncentracije ovih metala više u julu nego u septembru, što je posledica životnog ciklusa pčela. Izuzetak je primećen u Pančevu tokom 2014. godine, kada su koncentracije Cd, Co, Cu, Fe i Sr bile više u septembru u odnosu na jul. U Pančevu se može videti trend u redukciji koncentracije metala tokom godina. Al, Ba, Cr, Ni, Sr i Zn su pokazali značajne razlike između godina uzorkovanja, sa najvišim koncentracijama tokom 2013. godine, a najnižim tokom 2015. godine, što pokazuje da se stanje životne sredine, barem kada je reč o zagađenju metalima, poboljšava. U Vršcu ovo vii nije slučaj. Al, Cu, Ni i Sr su imali značajno niže koncentracije tokom 2014. godine u poređenju sa 2013. i 2015. godinom, što može biti posledica obilnih kiša i poplava tokom 2014. godine.

The increase of the human population on Earth has led to the increase of anthropogenic pollution. Toxic metals are considered one of the most dangerous pollutants because many of them are not biodegradable. With this in mind, it is important to find monitoring methods that cost less, cover bigger areas and give faster results. An alternative to classical monitoring techniques is the use of bioindicators. For the purposes of this study, honeybees (Apis mellifera L.) were used as bioindicators of toxic metal pollution in Serbia. One set of samples were adult honeybees collected during two sampling periods (July and September) in 2014 from apiaries located in Belgrade, Kostolac, and Mesić. Analyzed metals in these samples were Al, Ba, Cd, Co, Cr, Cu, Fe, Li, Mn, Na, Ni, Pb, Sr, and Zn. Another set of samples were honeybees from apiaries located in Pančevo and Vršac that were sampled during September 2013, July and September 2014, and July and September 2015. The bees were sampled during these two months because they represent the peak of bee activity. Metals analyzed in these samples were Al, Ba, Cd, Co, Cr, Cu, Fe, Mn, Na, Ni, Sr, and Zn. The same metals were analyzed in soil samples taken from Pančevo and Vršac in the vicinity of apiaries. For the quantification of analyzed metals, inductively coupled plasma – optical emission spectrometry (ICP-OES) was used. The two-way ANOVA showed that there are statistically significant differences between some of the analyzed metals in Belgrade, Kostolac, and Mesić. Significant differences between at least two of the locations were found for Al, Ba, Cr, Cu, Fe, Li, and Ni. Using principal component anlasys (PCA), cluster analysis, and ANOVA, it was determined that higher concentrations of Al, Fe, and Cr in Kostolac originate from thermal ix power plants “Kostolac A and B” and the accompanying ash disposal site. High concentrations of Cu in Belgrade are attributed to traffic and use of brakes that contain Cu. High Ni concentrations in Mesić are a consequence of farmers’ use of manure and artificial fertilizers, while higher concentrations of Ba and Cu at the same locations originate from spraying of orchards and vineyards in the vicinity of Mesić. Comparing metal concentrations between Vršac and Pančevo, it can be found that concentrations of Al, Ba, and Sr are significantly higher in Vršac. With the use of PCA, CA, and correlation tests between metals found in soil and those found in honeybees, it can be concluded that Ca, Cd, Na, Fe, Mn, and Zn have anthropogenic origin, and that their greatest sources are the “Hesteel Serbia Iron & Steel“ (former „Železara Smederevo“), thermal power plants “Kostolac A and B“, traffic, as well as intensive agriculture. Ca, Al, Mg, and Cr found in bees collected from the same locations are attributed to natural sources, while some of the elements (Cu, Ba, Sr, and Ni) have both anthropogenic and natural sources. During the 2014, samples were taken from all five locations: Belgrade, Pančevo, Kostolac, Vršac, and Mesić. ANOVA, PCA, and CA showed that concentrations of Ba and Cd are higher in Mesić compared to the other locations, which can be attributed to intensive agricultural activity in that region. Higher concentrations of Fe and Cr in Kostolac are the result of coal burning in thermal power plants “Kostolac A and B”, while significantly higher concentrations of Cu in Belgrade come from intense traffic. Seasonal variations in metal concentrations were observed for Al, Ba, Ca, Cd, Co, Cu, Fe, Mg, and Sr. In most cases, concentrations of these metals were higher in July compared to September, which is a consequence of the lifecycle of honeybees. Discrepancies were found for Pančevo during 2014, when concentrations of Cd, Co, Cu, Fe, and Sr were higher in September then in July. Comparing metal concentrations between the years, it was found that the trend in Pančevo is the reduction in their concentrations. For Al, Ba, Cr, Ni, Sr, and Zn, significant differences were observed between the years, where the highest concentrations were during x 2013, and lowest in 2015, which shows that the condition of the environment, when it comes to metal pollution, is improving. However, this is not the case in Vršac. Al, Cu, Ni, and Sr had significantly higher concentrations in 2014 compared to 2013 and 2015, where heavy rainfall during the peak of the bee activity in 2014 could be the reason for this.

Rasprostranjenje, diverzitet i struktura zajednica ektoparazita litofilnih slepih miševa (Chiroptera) centralnog Balkana

Distribution, diversity and structure of ectoparasite communities of cave-dweeling bats (Chiroptera) from the central Balkans

Studija predstavlja prvo sveobuhvatno istraživanje faune, brojnosti, učestalosti i specifičnosti različitih vrsta ektoparazita u odnosu na vrstu domaćina, kao i analizu ekoloških faktora koji utiču na brojnost i reprodukciju ektoparazita litofilnih slepih miševa na teritoriji centralnog Balkana. Trogodišnje terensko istraživanje je sprovedeno na 45 lokaliteta u Srbiji, Bosni i Hercegovini, Bivšoj Jugoslovenskoj Republici Makedoniji i Crnoj Gori. Prikupljeno je i analizirano preko 3500 jedinki ektoparazita klasifikovanih u 21 vrstu: tri vrste krpelja, sedam vrsta parazitskih grinja (uključujući jednog predstavnika familije Trombiculidae), osam vrsta parazitskih muva i tri vrste buva, od čega je najveći broj nalaza prvi za teritorije istraživanih država. Identifikovano je 75 asocijacija ektoparazit-domaćin. Veći broj vrsta ektoparazita slepih miševa identifikovane su kao specijalisti. Miniopterus schreibersii je domaćin sa najvećim brojem vrsta ektoparazita. Najrasprostranjenije vrste ektoparazita u studiji su bile Spinturnix psi i Penicillidia dufourii. Kod tri vrste slepih miševa (Miniopterus schreibersii, Myotis myotis i Rhinolophus ferrumequinum) utvrđeno je da postoji statistički značajan efekat pola domaćina na brojnost ektoparazita. Faza životnog ciklusa, pol, starost i reproduktivni status domaćina značajno su uticali na brojnost i reproduktivnu aktivnost vrsta ektoparazita karakterističnih za Miniopterus schreibersii. Osim originalnih podataka, studijom su obuhvaćeni i analizirani do sada objavljeni literaturni podaci o rasprostranjenju i domaćinima ektoparazita slepih miševa na istraživanoj teritoriji. ! !

The study represents first comprehensive research of fauna, abundance, prevalence and host specificity, as well as analysis of ecological factors influencing abundance and reproduction of ectoparasites parasitizing cave-dwelling bats at the territory of the central Balkans. Three years long field research was conducted at 45 localities in Serbia, Bosnia and Herzegovina, former Yugoslav Republic of Macedonia, and Montenegro. Over 3,500 ectoparasite specimens belonging to 21 species were identified: three species of ticks, seven species of mites (including one specimen of Trombiculidae family), eight species of bat flies and three species of fleas, most being the first records at the territory of investigated countries. A total of 75 host-parasite associations were identified. A high level of host specificity in analysed ectoparasite species was registered. Miniopterus schreibersii was host species with the greatest number of ectoparasite species. Mite Spinturnix psi and bat fly Penicillidia dufourii were the most widely distributed species. There was significant effect of host sex on ectoparasite abundance in three bat species (Miniopterus schreibersii, Myotis myotis and Rhinolophus ferrumequinum). Significant effects of host life cycle, age, sex and reproductive status on total number and reproductive activity were identified in the case of ectoparasite species characteristic for Miniopterus schreibersii. Besides original data, the study includes and analyses published literature data about ectoparasite hosts and distribution of bat ectoparasites at the studied territory. ! !

Rečna mrena (Barbus barbus) kao bioindikator zagađenja rečnih sedimenata metalima

Common barbel (Barbus barbus ) as a bioindicator of river sediment pollution with metals

Metali spadaju među značajne zagađivače akvatičnih ekosistema, s obzirom na njihovu stabilnost u ekosistemu i transport kroz lanac ishrane. Rečni sedimenti mogu deponovati visoke koncentracije metala, a brojni akvatični organizmi koriste sediment za različite potrebe u različitim fazama svog životnog ciklusa. U ovom radu analizirane su ukupna i dostupna frakcija 15 elemenata u površinskim sedimentima Dunava, Zapadne Morave i Južne Morave, u tri sezone 2012. godine (april, jul, novembar), na tri različite dubine sedimenta (0-5 cm, 5-10 cm, 10-15 cm), uz upotrebu optičke emisione spektrometrije sa induktivno spregnutom plazmom (ICP-OES). Koncentracije elemenata utvrđene su i u tkivima rečne mrene (Barbus barbus) uzorkovane iz sve tri analizirane reke, i to u mišiću, škrgama, jetri i crevu. Analiza koncentracije metala obuhvatila je i intestinalnog parazita Pomphorhynchus laevis (Acanthocephala) izolovanom iz mrene iz Dunava. Histološke analize urađene su u cilju utvrđivanja promena na škrgama i jetri mrene iz Dunava i Južne Morave. Za dodatnu analizu elemenata u sedimentu Zapadne Morave, u tkivima mrene iz Dunava, kao i u parazitu P. laevis, korišćena je skenirajuća elektronska mikroskopija sa energo- disperzivnom spektroskopijom (SEM-EDS). Statistička analiza i trofaktorska MANOVA pokazale su postojanje značajnih razlikau veličini čestica sedimenta, prosečnoj koncentraciji elemenata i sezonskim promenama u koncentraciji elemenata između tri posmatrane reke. Koncentracije Cu i Ni u Zapadnoj Moravi i Dunavu bile su veće od maksimalno dozvoljenih koncentracija (MDK) za sediment u Republici Srbiji, dok je koncentracija Hg bila veća od MDK u sve tri reke. Koncentracije Al, Ba, Cu, Fe, Mn i Zn varirale su između sezona u Zapadnoj Moravi i Dunavu, sa najvećim vrednostima u letnjem periodu. U Južnoj Moravi varirale su koncentracije Al, As, Fe, Mn i Zn sa najmanjim koncentracijama u letnjem periodu. Zapadna Morava je sadržala najveće količine finih čestica u svom sedimentu, dok se najveći deo sedimenta Južne Morave sastojao od čestica peska. SEM-EDS analiza sedimenta Zapadne Morave ukazala je na antropogeni izvor povećane koncentracije Cu vi u sedimentu ove reke. Analiza elemenata u tkivima mrene iz Zapadne Morave ukazala je na povećane koncentracije Cu i Ni u jetri i crevu, kao i Zn u mišiću mrene, što je u skladu sa koncentracijom ovih elemenata u sedimentu. SEM-EDS analiza organa mrene iz Dunava ukazala je na moguće antropogeno poreklo zagađivača u ovoj reci. Analizom koncentracije elemenata u P. laevis utvrđene su najveće koncentracije Cu, Zn i Pb, a SEM-EDS analiza je i u ovom slučaju ukazala na najverovatnije antropogeno poreklo ovih čestica. Histološka analiza jetre i škrga mrene iz Dunava i Južne Morave pokazala je statistički značajno veća oštećenja na ovim organima kod mrene iz Južne Morave, što je najverovatnije posledica organskog zagađenja ove reke.

Metals are listed among the most important pollutants in aquatic ecosystems due to their stability and mobility through the food webs. River sediments can contain high concentrations of metals, and various aquatic organisms use sediments for various requirements in different stages of their life cycle. In this research, total and bioavailable concentrations of 15 elements in surface sediments of three rivers – the Danube, the Zapadna Morava, and the Južna Morava, were analyzed during three seasons in 2012 (April, July, November), at three different sediment depths (0-5 cm, 5-10 cm, 10-15 cm), using inductively coupled plasma optical emission spectrometry (ICP-OES). Element concentrations were also determined in 4 tissues of the common barbel (Barbus barbus) sampled from all three rivers (muscle, gills, intestine, and liver). Element analysis was further carried out on barbel intestinal parasite Pomphorhynchus laevis (Acanthocephala), collected from the Danube River. Histological analyses of gills and liver from fish collected from the Danube and the Južna Morava was performed with the purpose of determining any tissue alterations. Scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM-EDS) was used for in-depth grain analysis of the Zapadna Morava sediments, as well as tissue and parasite P. laevis analyses of elements in barbel sampled from the Danube River. Statistical analysis and three-way MANOVA showed significant differences in sediment grain size, average element concentration, and seasonal element concentration in the three rivers. Concentrations of Cu and Ni in the Zapadna Morava River and the Danube River were above the maximum allowable concentrations (MAC) of elements in river sediments for the Republic of Serbia, while Hg concentration was above the MAC in all three observed rivers. Concentrations of Al, Ba, Cu, Fe, Mn, and Zn varied between seasons in the Zapadna Morava and the Danube, with the highest values during the summer season. Concentrations of Al, As, Fe, Mn, and Zn varied between seasons in the Južna Morava, with the lowest values during the summer season. Highest viii percentage of fine sediment particles was observed in the Zapadna Morava, while the highest percentage of sand was observed in the Južna Morava sediment. SEM-EDS analysis of the Zapadna Morava sediment pointed to the presence of anthropogenic source of the higher Cu concentration in this river compared to the other two rivers. Barbel collected from the Zapadna Morava River had higher concentrations of Cu and Ni in the liver and intestine, and Zn in the muscle tissue, which corresponded with the concentrations of these elements in the sediment. SEM-EDS tissue analysis of barbel specimens from the Danube River pointed to possible anthropogenic activities causing pollution in this river. Barbel intestinal parasite P. laevis had high concentrations of Cu, Zn, and Pb, and in this case SEM-EDS analysis confirmed possible anthropogenic sources of contamination. Histological analysis of barbel liver and gills collected from the Danube and the Južna Morava indicated statistically significant tissue alterations in fish collected from the Južna Morava River, most probably as a result of a high organic contamination of this river.

Između medija i instrumenta: savremena fenomenologija arhitekonskog modela

Contemporary Phenomenology of Architectural Model.

Ova doktorska disertacija ispituje kako se danas ispoljava pojavnost arhitektonskog modela, kroz pristup fenomenološke analize. Namera teze je dvojaka: da osvetli ulogu i značaj modela za projektovanje savremene arhitekture, i da istovremeno pokuša da razjasni potrebu za uvažavanjem fenomenologije u domenu savremenog arhitektonskog obrazovanja. Da bi se ovo sprovelo, rad je usredsređen na (1) pitanje savremenosti, (2) odnos fenomenologije i arhitekture i (3) analiziranje predmeta arhitektonskog modela. Teorijski plan poduprt je eksperimentalnim projektima modela, korišćenim kao arhitektonsko sredstvo za istraživanje autonomije identiteta, porekla forme i smisla svrhe, izrađenim kroz studentske vežbe na Arhitektonskom fakultetu u Beogradu. Ova teza takođe odražava i kako se sve mogu upotrebljavati modeli u arhitekturi.

This doctoral thesis explores the contemporary appearance of phenomenon of architectural model, accessed through phenomenological analysis. The intention is twofold: to highlight the role and importance of models for the design of contemporary architecture, and at the same time to clarify the need for recognition of phenomenology in the field of contemporary architectural education. To carry out these intentions, the work focuses on (1) the issue of contemporariness, (2) the open relationship of phenomenology and architecture and (3) the structural analysis of the object of architectural model. Theoretical plane is underpinned by experimental project-models, used as architecture’s investigative tool for the study of autonomy identity, origin of form and meaning of purpose, delivered through student exercises at Belgrade Faculty of Architecture. This thesis also presents reflections on how to use models in architecture.

Koncept integrisane rekreacije i mogućnosti primene u uslovima razvoja gradova Srbije

The concept of integrated recreation and possibilities of implementation in development of Serbian cities

Ova disertacija se bavi istraživanjem savremenog razvoja rekreacije i njene uloge u urbanom razvoju. U kontekstu traganja za novim obrascima razvoja, kojima bi se omogućilo prevazilaženje problema dezintegracije urbane strukture, rekreacija se preispituje kao integrativni urbani sadržaj koji je kao takav prepoznat u urbanoj teoriji. Fokus rada je na urbanoj rekreaciji, dakle onoj koja se odvija u gradu i odražava urbane stilove života. Pažnja je usmerena na istraživanje fenomena "integrisane rekreacije" za koji se pretpostavlja da značajno uvećava mogućnosti za rekreaciju u savremenom gradu. Ovaj fenomen je rezultat uključivanja rekreacije u različite procese proizvodnje prostora. Reč je o procesima koji, pored tradicionalnih normativno-distributivnih politika razvoja rekreacije u javnom sektoru, realizuju rekreativne sadržaje i prostore u savremenom gradu kao sredstvo razvoja grada. Cilj ovog istraživanja je bio da utvrdi osnove i ispita mogućnosti korišćenja rekreacije kao sredstva povezivanja urbane strukture, kao i da omogući primenu stečenih znanja u usmeravanju razvoja gradova Srbije. Budući da fenomen integrisane rekreacije nije uključen u postojeći koncept razvoja rekreacije u Srbiji, pretpostavka o promeni uloge rekreacije u razvoju savremenih gradova je uslovila da se, u skladu sa nalazima istraživanja, redefiniše i koncept rekreacije u urbanom razvoju u Srbiji. Zbog toga je i istraživanje bilo podeljeno u dva dela. U prvoj fazi su objašnjene okolnosti pojave fenomena integrisane rekreacije, sistematizovane karakteristike savremene provizije sadržaja i prostora rekreacije, i na osnovu teorijskog definisanja rekreacije kao integrativnog urbanog sadržaja ispitivane mogućnosti korišćenja rekreacije kao sredstva povezivanja urbane strukture. Ispitivanje karakteristika integrisane rekreacije i njenog integrativnog potencijala u savremenom gradu bilo je zasnovano na želji da se prepoznaju različite mogućnosti razvoja kroz rekreaciju, kao i da se pri tom sagledaju vrednosti koji leže u njihovoj osnovi. Zbog toga je fokus sa direktog izbora reprezentativnih primera iz prakse, pomeren na istraživanje aktuelnih koncepata razvoja grada kroz rekreaciju, koji su tu praksu omogućili ili su na osnovu nje bili generisani. Reč je o konceptima „grada zabave“, „grada fantazije“, „grada turizma“, „grada doživljaja“, „grada igre“, „zdravog/aktivnog grada“, „zelenog grada“ i „kreativnog grada“. Analizom ovih koncepata uspostavljene su veze između razvojnih problema, aktera proizvodnje prostora, formi rekreacije koje proizvode i veza u urbanoj strukturi koje uspostavljaju. Identifikovane su i sistematizovane karakteristične forme i nivoi razvoja integrisane rekreacije, kao i lokacije na kojima različiti procesi proizvodnje prostora integrišu rekreaciju u urbani prostor. Uporednom analizom su ispitivane sličnosti i razlike pojedinih koncepata i identifikovane prioritetne lokacije za razvoj kroz rekreaciju. Utvrđeno je da „integrisana rekreacija“ poseduje sopstvenu logiku vezivanja za prostor koja prati različite razvojne potrebe grada, kao i da različiti koncepti razvoja kroz rekreaciju pružaju različite mogućnosti lociranja, forme i nivoe razvoja prostora rekreacije, i omogućavaju različite vrste rekreativnih praksi. Uključenje rekreacije u različite procese proizvodnje prostora kroz različite koncepte razvoja na taj način uvećava mogućnosti za integrativne rekreativne prakse i tako doprinosi povezivanju urbane strukture u savremenom gradu. Na osnovu rezultata istraživanja u prvom delu rada, sagledavanja specifičnih uslova razvoja gradova Srbije, kao i preispitivanja postojećeg koncepta rekreacije, definisane su nove vrednosne i sadržajne osnove razvoja rekreacije u usmeravanju razvoja gradova Srbije. U okviru novog Koncepta integrisane rekrecije razvijen je Model integrisanja urbane strukture razvojem rekreacije. Ovaj model povezuje proces proizvodnje prostora integrisane rekreacije, specifične situacije integrisane rekreacije i mogućnosti povezivanja urbane strukture. Razmotrene su mogućnosti operacionalizacije i implikacije rezultata istraživanja u odnosu na urbani dizajn, planiranje i upravljanje i definisane pretpostavke za primenu koncepta integrisane rekreacije u razvoju gradova Srbije. Na primeru Beograda je proveravana mogućnost primene koncepta integrisane rekreacije u lokalnom kontekstu. Na osnovu analize strateških dokumenata razvoja grada i inicijalnog razmatranja primera iz prakse, utvrđen je relativno visok nivo razvijenosti fenomena integrisane rekreacije u Beogradu, kao i ograničene mogućnosti koje postojeći strateški dokumenti pružaju za korišćenje rekreacije kao sredstva razvoja grada. Analiza uloge rekreacije u razvoju centralnog gradskog priobalja Save je ukazala da je u postojećim uslovima razvoj kroz rekreaciju moguć i u Beogradu, ali kroz manje fragmente i taktičke intervencije.

This thesis explores the contemporary urban recreation and its use as a tool for urban development. Based on integrative capacity of recreation as an urban activity, the aim of the research was to examine possibilities to use this asset in development of Serbian cities. Since the existing concept of recreation development in Serbia doesn‟t acknowledge its possible use as a tool for urban development, thesis aims to redefine the concept of recreation so that it can be implemented in Serbian context. For that reason the research was organized on two levels. At the first level the integrative role of recreation was examined through research in urban theory and in the fields of psychology, sociology and geography of leisure. Integrative capacity of urban recreation was defined through concepts of leisure experience, encounter and practice and as such operationalized as fluid, focal and porous integrative practice. Different possibilities for activating integrative potential of urban recreation were identified for autonomous and integrated forms of urban recreation. The occurrence and spreading of the phenomena of integrated recreation is then explained and its integrative capacity explored through comparative analysis of eight concepts of development through recreation: „city of fun“, „fantasy city, „tourist city“, „experience city“, „city of play“, „healthy/active city“, „green city“ and „creative city“. The aim was to identify possibilities and constraints of each mode of urban development. The characteristics of each concept of development through recreation were mapped on hypothetical model of contemporary city so that a new „landscape of integrated recreation“ can be identified. The results have proved that various developent paths enabled recreation to function as an integrative tool for urban development at different levels, in various forms and on many locations in the city. These results formed the basis for redefining a concept of recreation in Serbia and for developing a model for integrating urban structure through development of recreation opportunities in urban space. This conceptual model connects the process of production of recreational space, characteristics of recreational spatial situations and their possible integrative effects on urban structure. The new concept of integrated recreation was developed by taking into account specificis of Serbian cities and particular social, economical, political and cultural conditions in which they develop. The implementation of the concept of integrated recreation was explored by using the city of Belgrade as a case study. The content analysis of four main planning documents and stategies for city development showed that basis for using recreation as a tool for urban development exists, but that there are many restrictions for the implementation. At the other hand, we have shown that in practice various forms of integrated recreation can be identified, but that there are not connected with official planning system. Finnaly, the analysis of the role that integrated recreation played in re-development of Belgrade central waterfront on the Sava river showed that through spatial fragments and in form of urban tactic, recreation can be used as a tool for urban developemnt in Serbian cities.

Geometrijsko-morfometrijske analize glavenog skeleta šakala (Canis aureus) i lisice (Vulpes vulpes) sa područja Srbije: biogeografski aspekti morfološke varijabilnosti

Geometric-morphometric analyses of golden jackal (Canis aureus) and red fox (Vulpes vulpes) skulls from the territory of Serbia: biogeographical aspects of morphological variability

Metodama geometrijske morfometrije, analizirana je varijabilnost veličine i oblika ventralnog, dorzalnog i lateralnog kranijuma i mandibule subadultnih i adultnih šakala (Canis aureus) i lisice (Vulpes vulpes) iz tri biogeografske oblasti sa područja Srbije: panonske, planinsko-kotlinske i peripanonske. Kod obe uzrasne grupe, utvrđeno je odsustvo razlika u veličini i prisustvo razlika u obliku glavenog skeleta između šakala iz različitih biogeografskih oblasti. Na većoj geografskoj skali ventralni, a na manjoj lateralni kranijum se pokazao kao najinformativniji. Između lisica iz tri biogeografske oblasti nema razlika u veličini, ali postoje razlike u obliku glavenog skeleta, sa lateralnim kranijumom kao najinformativnijim. Oblik glavenog skeleta lisica varira duž gradijenata geografske širine i dužine, kao i u zavisnosti od udela agrarnih površina, za razliku od njegove veličine koja je uniformna. Promene oblika ventralnog i lateralnog kranijuma uslovljene razlikama u udelu agrarnih površina su u saglasnosti sa promenama oblika duž gradijenata geografske širine i dužine, kao i sa biogeografskim obrascima varijabilnosti oblika. Kod obe vrste, statička alometrija ne utiče na biogeografske obrasce varijabilnosti oblika glavenog skeleta. Polni dimorfizam u veličini glavenog skeleta je izraženiji kod lisica u odnosu na šakale. Kod šakala, polni dimorfizam u obliku najuočljiviji je na nivou ventralnog kranijuma i mandibule. Kod lisica, polni dimorfizam u obliku, detektovan na nivou svih kranijalnih perspektiva, najuočljiviji je na nivou lateralnog kranijuma. Kod obe vrste, razlike između polova u obliku baze lobanje su uslovljene razlikama u veličini, dok širi rostralni region i zigomatični lukovi kod mužjaka nisu u vezi sa promenama u veličini.

Geometric morphometric approaches were employed to explore size and shape changes of ventral, dorsal and lateral cranium and mandible in subadults and adults of golden jackal (Canis aureus) and red fox (Vulpes vulpes) from three biogeographical areas in Serbia: pannonian, mountain-valley and peripannonian. In both age groups, absence of size, but presence of cranial and mandibular shape differences were found between jackals from different biogeographical areas. At larger geographic scale ventral cranium was the most informative, while at smaller geographic scale it was lateral cranium. There were no skull size differences among foxes from three biogeographical areas, whereas shape changes were detected, and lateral cranium was the most informative. While uniform considering size, the shape of red fox skulls varies in relation to latitude and longitude, as well as depending on proportion of agricultural habitats. Shape changes of ventral and lateral cranium related to the proportion of agricultural habitats correspond to those associated with latitudinal and longitudinal gradients resembling biogeographical patterns of shape changes. Static allometry does not influence biogeographical patterns of shape changes observed for both species. Skull size differences between the sexes were more pronounced in foxes than in jackals. In jackals, sexual shape dimorphism was the most obvious at the level of ventral cranium and mandible. In foxes, sexual dimorphism was also detected for cranial shape, and lateral cranium was the most informative. In both species, basicranial shape changes between the sexes are size-related, while the wider rostral and zygomatic regions in males are not associated with size differences.

Morphological and molecular characterization of Aphidius eadyi species complex (Hymenoptera, Braconidae, Aphidiinae), parasitoids of pea aphid - Acyrthosiphon pisum Harr. (Hemiptera, Aphididae)

Морфолошка и молекуларна карактеризација врста Aphidius eadyi комплекса (Hymenoptera, Braconidae, Aphidiinae), паразитоида зелене луцеркине ваши - Acyrthosiphon pisum Harr. (Hemiptera, Aphididae)

Acyrthosiphon pisum Harris je jedna od ekonomski najznačajnijih biljnih vaši u poljoprivredi, prvenstveno na kulturama iz familije Fabacae. Iz tog razloga su i brojni programi biološke kontrole realizovani širom sveta. U ovim programima su veoma često kao agenti korišćene vrste Aphidius eadyi kompleksa. I pored česte upotrebe i značaja, broj vrsta unutar kompleksa, njihova taksonomija i rasprostranjenje su uglavnom nepoznati. U ovoj studiji izvršena je identifikacija svih parazitoida iz potfamilije Aphidiinae koji parazitiraju A. pisum na prostoru Evrope. Među njima su, upotrebom morfoloških i molekularnih analiza (sekvenci mtCOI gena) identifikovane tri vrste koje pripadaju Aphidius eadyi kompleksu: Aphidius smithi, A. eadyi i A. banksae. Morfološkom karakterizacijom je utvrđeno da su za razlikovanje ovih vrsta najznačajniji sledeći morfološki karakteri: oblik brazdi na anterolateralnom regionu petiolusa, oblik centralne areole na propodeumu, oblik i nervatura prednjih krila. Oblik prednjih krila je analiziran upotrebom geometrijske morfometrije i utvrđeno je da se sve tri vrste razlikuju i pored manjeg preklapanja. Morfološke razlike su potvrđene i molekularnim analizama kojima je utvrđeno da se genetičke distance između vrsta A. eadyi kompleksa kreću u rasponu od 5% do 7,4%. Identifikacija vrste Aphidius banksae, kao široko rasprostranjenog parazitoida zelene lucerkine vaši predstavlja najinteresantniji nalaz ove studije. Utvrđeno je da rasprostranjenje vrste A. banksae obuhvata najveći deo zapadnog Palearktika, od Ujedinjenog Kraljevstva do Izraela. Dodatno, dat je i ponovni opis vrste Aphidius banksae kao i ključ za identifikaciju svih parazitoida potfamilije Aphidiinae koji parazitiraju Acyrthosiphon pisum u Evropi.

Acyrthosiphon pisum Harris is an aphid species of the greatest agricultural importance. It is a major pest on several plants of the family Fabacae, and there have been numerous programs involving biological control of Acyrthosiphon pisum worldwide. Species belonging to the Aphidius eadyi group have been used as biocontrol agents in those programs, but knowledge about their taxonomy and distribution has remained scarce with big gaps. Here we identify all aphidiine parasitoid species that have parasitized A. pisum in Europe, including three species within the Aphidius eadyi species group, using both molecular (mtDNA COI sequences) and morphological analyses. The Aphidius eadyi species group consists of the following species: Aphidius smithi, A. eadyi, and A. banksae. Morphological characterization showed that the most important morphological characters for separation of species of the Aphidius eadyi group are: shape of costulae on the anterolateral part of the petiole; shape of the central areola on the propodeum; and shape and venation of the forewings. Forewing shape was analysed using geometric morphometrics, and it is demonstrated that all three species differ in wing shape with some overlap. Morphological differences were confirmed by molecular data, mean genetic distances between the species varying from 5 to 7.4%. Identification of Aphidius banksae as a widely distributed pea aphid parasitoid whose range covers most of the western Palaearctic (from the United Kingdom to Israel) is the most interesting finding of this study. In addition, Aphidius banksae is diagnosed and redescribed. A key for identification of all aphidiine species attacking Acyrthosiphon pisum in Europe is provided.

Imunomodulatorni efekti antikoagulanta varfarina kod pacova

Immunomodulatory effects of anticoagulant warfarin in rats

Varfarin (4-hidroksikumarin) je antagonist vitamina K. Ovaj kumarinski derivat se koristi kao antikoagulantni rodenticid i kao terapeutsko sredstvo u profilaksi tromboembolijskih bolesti. Antikoagulantni efekat varfarina se zasniva na inhibiciji koraka, koji zavise od vitamina K, u sintezi brojnih faktora koagulacije u jetri. Zahvaljujući inhibiciji ciklusa vitamina K, varfarin utiče i na druge proteine neophodne za biološke procese van hemostaze (rast i kalcifikaciju kostiju, rast glatkomišićnih ćelija vaskulature i mezangijskih ćelija i druge), dovodeći do štetnih posledica. Varfarin inhibira i procese koji nisu u vezi sa vitaminom K, uključujući rast tumora. Malobrojni podaci ukazuju da ovaj agens može da deluje i na pojedine aspekte imunske funkcije. Mehanizmi delovanja varfarina na imunski sistem su, međutim, najmanje poznati. Ova disertacija je imala za cilj da ispita imunomodulatorni potencijal varfarina nakon njegove epikutane i oralne primene kod pacova. Akutni epikutani tretman (tri dana za redom u dozi od 10 μg i 100 μg natrijum varfarina) odgovara profesionalnoj ili akcidentalnoj izloženosti, a subakutni oralni tretman (30 dana u dozi od 0.35 mg/L i 3.5 mg/L natrijum varfarina u pijaćoj vodi) odgovara terapijskoj primeni. Proinflamatorni i imunomodulatorni potencijal varfarina je ispitan analizom humoralnih parametara (koncentracija akutno faznih proteina i proinflamatornog citokina, interleukina 6 (IL-6) u plazmi, kao i aktivnosti osnovnih enzima antioksidativne odbrane superoksid dismutaze i katalaze u eritrocitima), kao i ćelijskih parametara [kvantitativne i kvalitativne promene leukocita periferne krvi, posebno polimorfonuklearnih leukocita (PMN)] kao pokazatelja zapaljenja na sistemskom nivou. Mehanizmi dejstva na polimorfonuklearne ćelije periferne krvi ispitani su u pogledu njihove oksidativne aktivnosti, produkcije proinflamatornih citokina, faktora nekroze tumora (TNF) i IL-6, sposobnosti adhezije, migracije i ekstravazacije u ciljna tkiva, korišćenjem modela subkutano implantiranih sterilnih polivinilskih sunđera. Rezultati su pokazali da varfarin dospeva u organizam i kada se nanosi na kožu i nakon oralne primene, sudeći po produženju protrombinskog vremena. Epikutana aplikacija ne dovodi do promena koncentracije proteina akutne faze i IL-6, ali je zapažen porast aktivnosti katalaze pri višoj dozi varfarina. U ovom režimu primene varfarina zapažene su kvantitativne promene leukocita periferne krvi (povećanje broja neutrofilnih i eozinofilnih leukocita uz istovremeno smanjenja broja limfocita). Evidentne su i promene karakteristika eritrocita. Varfarin nije ispoljio direktno citotoksično dejstvo na polimorfonuklearne leukocite, ali je doveo do promena njihove aktivnosti uključujući: (1) povećanje broja ćelija koje iskazuju CD11b (pri obe doze) bez efekta na adheziju, (2) diferencijalni efekat na oksidativnu aktivnost polimorfonukleara [povećanje mijeloperoksidaze i povećanu sposobnost redukcije tetrazolijumove soli nakon stimulacije lipopolisaharidom (LPS) (pri višoj dozi), ali smanjenje nestimulisane produkcije azot-oksida (pri obe doze) i nepromenjenu produkciju nakon stimulacije LPS-om] i (3) smanjenje nestimulisane i LPS-om stimulisane produkcije i ekspresije proinflamatornih citokina TNF i IL-6 (kao rezultat inhibicije na transkripcionom nivou). Oralna primena varfarina je dovela do uspostavljanja sistemske zapaljenjske reakcije koju su karakterisale povećanje proteina akutne faze i IL-6 (pri obe doze), kao i promene aktivnosti superoksid dismutaze i katalaze. Viša doza varfarina je dovela do povećanja broja neutrofilnih leukocita, ali do smanjenja broja limfocita. Varfarin nije bio direktno toksičan za polimorfonuklearne leukocite, ali je uticao na kvalitativne promene ovih ćelija. One su obuhvatile smanjenje broja ćelija koje iskazuju CD11b (pri obe doze), povećanje svih aspekata oksidativnih aktivnosti ovih ćelija, ali sa diferencijalnim efektom na njihov citokinski odgovor (sniženje TNF pri višoj dozi varfarina, kao rezultat inhibicije na transkripcionom nivou, bez uticaja na IL-6). I pored povećanja broja polimorfonuklearnih leukocita koji su migrirali u sunđer, zapažen je slab efekat na aktivnost ovih ćelija, što je ukazalo na značaj sistemskog okruženja u ostvarivanju uticaja na ćelijsku aktivnost. Rezultati ove studije su pokazali imunomodulatorni kapacitet varfarina čime se proširuje lista bioloških aktivnosti varfarina koje nisu vezane za hemostazu, a što ukazuje na potrebu za razmatranjem štetnih efekata varfarina i dodatnih ispitivanja efekata ovog agensa. Ovo može da bude važno za kritičko sagledavanje rizika od profesionalne izloženosti, a posebno kliničke primene varfarina (kao leka), ako se ima u vidu da se ovaj agens, i pored pojave novih oralnih antikoagulanata sa direktnim inhibitornim dejstvom na faktore koagulacije IIa i Xa, i dalje svrstava u one koji se najčešće prepisuju kao antikoagulantna terapija.

Warfarin (4-hydroxycoumarin) is a vitamin K antagonist. This coumarin derivative is used as an anticoagulant rodenticide and as a therapeutic agent for the prophylaxis of thromboembolic disorders. The anticoagulant effect of warfarin is based on the inhibition of the steps which are dependent on Vitamin K in the synthesis of a number of coagulation factors in the liver. Owing to the inhibition of the vitamin K cycle, warfarin also affects the other proteins which are necessary for the biological processes apart from hemostasis (growth and calcification of bones, vascular smooth muscle cells, mesangial cells and others), leading to harmful effects. Warfarin also inhibits the processes which are not related to vitamin K, including the growth of tumors. Few data suggest that this agent may act on certain aspects of immune function. Mechanisms of action of warfarin on the immune system are, however, the least known. This study was aimed to investigate the immunomodulatory potential of warfarin after its epicutaneous and oral administration in rats. The acute epicutaneous treatment (within three consecutive days in doses of 10 μg and 100 μg sodium warfarin) corresponds to professional or accidental exposure, and the subacute oral treatment (30 days in doses of 0.35 mg/L and 3.5 mg/L sodium warfarin in drinking water) corresponds to the therapeutic use. The proinflammatory and immunomodulatory potential of warfarin has been tested by analyzing the humoral parameters (concentration of acute phase proteins and pro- inflammatory cytokines, interleukin 6 (IL-6) in the plasma, as well as the activities of basic enzymes of antioxidant defence, superoxide dismutase and catalase in erythrocytes) as well as the cell parameters [quantitative and qualitative changes in peripheral blood leukocytes, especially polymorphonuclear leukocytes (PMN)] as an indicator of inflammation at the system level. Mechanisms of effects to the peripheral blood polymorphonuclear cells were tested with regard to their oxidative activity, production of pro-inflammatory cytokines, tumor necrosis factor (TNF) and IL-6, ability of adhesion, migration and extravasation in the target tissues, using models of subcutaneously implanted sterile polyvinyl sponges. The results showed that warfarin comes into the organism both when it is applied on the skin and after oral administration, judging by the extension of prothrombin time. Epicutaneous application does not lead to changes in the concentration of acute phase proteins and interleukin 6, but it has been noticed a significant increase of catalase activity at the higher dose of warfarin. In this mode of application of warfarin quantitative changes were observed in peripheral blood leukocytes (increase in the number of neutrophil and eosinophil leukocytes with a simultaneous decrease in the number of lymphocytes). There are obvious changes in the characteristics of red blood cells. Warfarin did not exhibit a direct cytotoxic effect on polymorphonuclear leukocytes, but it led to changes in their activities, including: (1) increasing the number of cells that express CD11b (at both doses) but with no effect on adhesion, 2) a differential effect on the oxidative activity of polymorphonuclears [increase in myeloperoxidase and an enhanced ability to reduce a tetrazolium salt after stimulation with lipopolysaccharide (LPS) (at a higher dose), but decrease in non-stimulated production of nitric oxide (at both doses) and an unchanged production upon stimulation with LPS] and (3) reduction of the spontaneous and LPS-stimulated production and the expression of pro-inflammatory cytokines TNF and IL-6 (as a result of inhibition at the transcriptional level). Oral administration of warfarin led to the systemic inflammatory reaction which was characterized by the increase in acute phase proteins and IL-6 (at both doses), and change in activities of superoxide dismutase and catalase. A higher dose of warfarin led to the increase in the number of neutrophil leukocytes, but to the decrease in the number of lymphocytes. Warfarin was not directly toxic to the polymorphonuclear leukocytes, but it influenced the qualitative changes in these cells. These included a reduced number of cells that express CD11b (at both doses), increase of all aspects of the oxidative activity of these cells, but with differential effects on their cytokine response (reduced TNF at the higher dose of warfarin, as a result of inhibition at the transcriptional level, without affecting the IL-6). Despite the increased number of polymorphonuclear leukocytes which migrated to the sponge, it was noticed a poor effect on the activity of these cells, which indicated the importance of the system environment in influencing the cell activity. Results of this study have shown the immunomodulatory capacity of warfarin thus extending the list of biological activities of warfarin that are not related to hemostasis, which indicates the need to consider the harmful effects of warfarin and to do further research on the effects of this agent. This may be important for the critical evaluation of the risk of occupational exposure, and specifically a clinical use of warfarin as a medicine, if we have in mind that this agent, despite the emergence of new oral anticoagulants with a direct inhibitory effect on coagulation factors IIa and Xa, is still classified among those most often prescribed as an anticoagulant therapy.

Potencijal rekombinantnih proteina pljuvačke krpelja Ixodes ricinus (Linnaeus, 1758) za detekciju specifičnih antitela kao markera uboda krpelja

Potential of recombinant Ixodes ricinus (Linnaeus, 1758) tick saliva proteins for detection of specific antibodies as markers of tick bite

Procena izloženosti krpeljima je značajna u epidemiološkim studijama oboljenja čije uzročnike prenose krpelji, a potvrda uboda krpelja je važna za njihovu dijagnostiku. U ovom radu su ispitana tri proteina pljuvačke, PA107, AV422 i kalretikulin (CAL), krpelja Ixodes ricinus, medicinski najznačajnije vrste u Evropi, u kontekstu upotrebe za detekciju specifičnih antitela u serumu domaćina, kao markera uboda. Analiza varijabilnosti njihovih sekvenci je ukazala na visoku konzerviranost sva tri proteina, pri čemu se PA107 pokazao kao najkonzerviraniji. S obzirom na ubikvitarnu zastupljenost CAL kod eukariota, dalji fokus u ovoj studiji je bio na PA107 i AV422, kao proteinima koji su specifični za krpelje i čiji homolozi su prvi put analizirani kod vrste Ixodes ricinus. Ispitivana je reaktivnost seruma eksperimentalnih životinja, pacova, na kojima su hranjene larve Ixodes ricinus ili Dermacentor reticulatus, sa rekombinantnim formama IrPA107 i IrAV422. Korišćenjem rIrAV422, specifična antitela su detektovana kod svih infestiranih pacova, što ukazuje da se rIrAV422 može upotrebiti za detekciju markera uboda različitih grupa tvrdih krpelja uz visoku senzitivnost i specifičnost. Reaktivnost u slučaju IrPA107 je bila značajno manja. Za ispitivanje upotrebe rIrAV422 u detekciji markera uboda u serumima domaćina izloženih krpeljima u prirodnom okruženju, korišćeni su lovački psi kao adekvatna indikatorska grupa. Dobijena seroreaktivnost kod svih analiziranih pasa u studiji ukazuje na širok potencijal primene ovog proteina pljuvačke u proceni izloženosti različitim vrstama krpelja i rizika od infekcije patogenima koje oni prenose.

Assessment of exposure to ticks is a significant data in epidemiological studies of tick-borne diseases, and confirmation of tick bite is important for their diagnosis. In this study, three saliva proteins, PA107, AV422 and calreticulin (CAL), of Ixodes ricinus tick, medically most important species in Europe, were examined in the context of their use for detection of specific antibodies in sera of hosts, as bite markers. Sequence variability analysis revealed high conservativity of all three proteins, with PA107 as the most conservative. Since CAL is ubiquitous in eukaryotes, the further focus of this study was on PA107 and AV422, as proteins that are specific for ticks. Homologues of these proteins from Ixodes ricinus were analyzed for the first time. Reactivity of sera of experimental animals, rats, which were infested separately with Ixodes ricinus or Dermacentor reticulatus larvae, with recombinant forms of IrPA107 and IrAV422 were examined. Specific antibodies were detected in sera of all infested rats using rIrAV422, which implied its usage for detection of bites of tick species that belong to different groups of ixodid ticks, with high sensitivity and specificity. Reactivity with IrPA107 was significantly lower. Hunting dogs, as good sentinels, were used for assessment of rIrAV422 usage for detection of markers in sera of hosts which are exposed to ticks in natural environment. Obtained seroreactivity in all dogs implies broad applicability of this saliva protein, in terms of assessment of exposure to different tick species and risk of infection with tick-borne pathogens.

Diverzitet i uloga mikromiceta u procesu biodeterioracije zidnih slika crkve Svetog Vaznesenja Gospodnjeg u Velikom Krčimiru

Diversity and role of micromycetes in wall painting biodeterioration process: Church of the Holy Ascension in Veliki Krčimir

Analiziran je diverzitet i sezonska dinamika mikromiceta na zidnim slikama i u vazduhu istraživane crkve. Identifikovana je mikobiota od 46 taksona mikromiceta, sa dominacijom Aspergillus, Penicillium i Cladosporium vrsta. Najveći diverzitet je zabeležen tokom leta, dok je najveća brojnost propagula po jedinici površine (“fungalni otisak”) konstatovana u zimu. Aeromikobiota je okarakterisana sa 33 taksona gljiva. Kontaminacija vazduha propagulama gljiva tokom godine višestruko je prevazilazila standarde za zatvorene prostore. Mikroskopska analiza biofilma pokazala je da su reproduktivne strukture Cladosporium sp. i Chaetomium sp. u kontaktu bojenog sloja i maltera glavni biotski faktor deterioracije. Metodom ATP bioluminiscenije, 75% površina zidnih slika je okarakterisano kao “Zona opasnosti”. Monitoringom indukovane i spontane kolonizacije na modelu zidne slike zaključeno je da su lihenizovane i mikrokolonijalne gljive uzrok fenomena “biopitting”. Testirani izolati su demonstrirali veliki potencijal deterioracije zidnih slika u eksperimentima in vitro. BAC i novosintetisani BAC/FNP nanokompozit pokazali su dobru antifungalnu aktivnost, kao i uticaj na povećanje produkcije aflatoksina B1 i smanjenje produkcije ohratoksina A. Etarsko ulje tamjana imalo je jači antifungalni efekat u odnosu na etarsko ulje smirne, ali slabiji u poređenju sa smešom ulja. Dim tamjana poseduje inhibitorno dejstvo na germinaciju konidija, mehaničkim i hemijskim dejstvom voštanog sloja deponovanog iz dima, in vitro. Tretman vazduha crkve dimom tamjana redukovao je nivo kontaminacije vazduha fungalnim propagulama za približno 80%. Preko predloženog metodološkog protokola, data je mogućnost implementacije rezultata ovog istraživanja u praksu konzervacije i restauracije kulturnih dobara.

Diversity and seasonal dynamics of micromycetes from wall paintings and surrounding air of investigated church was analyzed. A total of 46 fungal taxa was documented, with dominance of Aspergillus, Penicillium and Cladosporium species. The greatest species diversity was recorded in the summer, while the highest abundance of fungal propagules per unit area (“fungal print”) was observed in the winter. Air mycobiota was characterized by 33 fungal taxa. Fungal air contamination, throughout the year, exceeded permisable levels for indoor environment. Microscopic biofilm analyzes revealed that reproductive structures of Cladosporium sp. and Chaetomium sp. are the main biodeteriogens. ATP bioluminescence method showed that 75% of the surfaces are deemed as “Danger zone”. Monitoring of induced and spontaneous colonization of mock-model established lichenized and micro-colonial fungi as the main causative agents of “biopitting” phenomenon. Tested isolates demonstrated pronounced deterioration capabilities in experiments in vitro. BAC and newly- synthesized BAC/FNP nanocomposite had very strong antifungal activity, stimulated aflatoxin B1 production, and inhibited ochratoxin A production. Frankincense essential oil showed stronger antifungal property compared to the oil obtained from myrrh, but weaker than the oil mixture. Burn incense had inhibitory effect on conidia germination via mechanical and chemical activity of thin waxy layer deposited from fume, in vitro. Treatment of air within church, with burn incense, reduced the level of fungal air contamination by approximatly 80%. Through the proposed methodological protocol, the possibility of implementing results into the practice of conservation and restoration of cultural heritage is given.

Razvoj visoko automatizovanih mikrotitarskih testova na Saccharomyces cerevisiae za detekciju jedinjenja sa antifungalnim i citostatskim dejstvom

Development of two high-throughput screening methods to identify molecules with antifungal and cytostatic activity using Saccharomyces cerevisiae

Kvasac Saccharomyces cerevisiae je široko upotrebljavan model za istraživanje potencijala malih molekula u modulaciji signalnih puteva relevantnih za nastanak i razvoj bolesti u eukariotskim organizmima. U ovoj tezi je opisan razvoj dva visoko automatizovana testa zasnovana na praćenju ćelijske vijabilnosti kroz merenje fluorescentnog signala redukcijom alamarBlue® (resazurina) ili direktnim merenjem optičke gustine. Obe metode su pogodne za pretragu velikog broja uzoraka, u mikrotitarskim pločama sa 348 i 1536 bunarčića upotrebom diploidnog pdr5/pdr5 soja S.cereviseae kome nedostaje glavni transmembranski transporter Pdr5p, koji doprinosi multirezistenciji na lekove. Oba testa su pokazala visok kvalitet sa Z’ vrednosti > 0,5. Korišćenjem alamarBlue® testa ispitano je 1.101.408 jedinjenja male molekulske težine, od kojih je 25.144 inhibiralo rast kvasca. Određivanjem IC50 vrednosti, od 1930 odabranih jedinjenja iz primarnog testiranja, inhibitorni potencijal je potvrđen u 868 (45%) u oba ili bar jednom testu. Rezultat dobijeni haplodeficijentnim profilisanjem genoma kvasca su pokazali njihovu visoku raznovrsnost delovanja, 572 odbrana aktivna jedinjenja je uticalo na rast >25% ukupnog broja ispitivanih sojeva kvasca. Molekuli sa inhibitornom aktivnošću detektovanom na ovakav način predstavljaju impresivnu bazu za dalje istraživanje mehanizama putem kojih suprimiraju proliferaciju i/ili vijabilnost ćelija kvasaca, drugih patogenih gljiva, ali i sisarskih ćelija.

The budding yeast S. cerevisiae is widely used model organism to elucidate the mechanism of action of low molecular weight compounds in modulation of signaling pathways involved in different diseases in eukaryotic organisms. This thesis describes the development of two high throughput screening methods based on cell viability either by monitoring the reduction of alamarBlue® (resazurin) or by direct optical measurement of cell growth. Both methods can be miniaturized to allow screening of large numbers of samples, in 384 and 1536-well format using the diploid pdr5/pdr5 strain of S.cereviseae lacking the major transmembrane multi-drug resistance pump Pdr5p. Both tests show high quality with Z’ value > 0.5. With the alamarBlue® approach, 1,101,408 low molecular weight compounds were screened, identifying 25,144 primary hits. Dose response curves (IC50) generated for a subset of 1930 active compounds using both alamarBlue® and optical density methods confirmed inhibitory potential for 868 of them either by both or one method. In genome-wide haploinsufficiency profiling (HIP), 572 of selected hits demonstrated a diverse mechanism of action, affecting >25% of all yeast strains. These hits represent an impressive basis for further evaluation of mechanisms of action for suppression of yeast’s, pathogenic fungi and mammalian cells proliferation and viability.

Morfološka varijabilnost, evolucija i razviće kičmenice kod velikih mrmoljaka (Triturus, Salamandridae, Caudata)

Morphological variability, evolution and development of vertebral column in crested newts (Triturus, Salamandridae, Caudata)

Analize intra- i interspecijske varijabilnosti u ontogeniji morfoloških struktura (npr. razlike u vremenu diferencijacije, stopi i pravcu ontogenetskih promena u veličini i obliku), kao i varijabilnosti na adultnom stupnju, omogućavaju utvrđivanje obrazaca morfološke varijabilnosti i sagledavanje procesa i mehanizma koji dovode do evolucionih promena morfoloških struktura. U ovoj tezi izučavana je morfološka varijabilnost osovinskog skeleta u okviru monofiletske grupe velikih mrmoljaka koju čine devet vrsta roda Triturus. Različit stepen izduživanja tela, varijabilnost u broju trupnih pršljenova kao i poznati filogenetski odnosi čine ovu grupu izuzetno pogodnim model-organizmima za studije razvića kičmenice i evolucije kičmenih pršljenova. Histološkim metodama praćeno je postembrionsko razviće trupnih pršljenova, radi utvrđivanja eventualnih razlika u vremenu formiranja, osifikacionim sekvencama kao i stopi njihove diferencijacije. Za ove analize odabrane su dve vrste velikih mrmoljaka koje se razlikuju u stepenu izduživanja tela, odnosno u broju trupnih pršljenova. Utvrđeno je da nema razlika u vremenu i načinu formiranja pršljenova između vrsta. Tokom formiranja kičmenih pršljenova zabeležena su tri tipa osifikacije: 1) perihordalna osifikacija, 2) perihondrijalna osifikacija i 3) endohondralna osifikacija. Takođe, zabeleženo je i prisustvo intravertebralne (notohordalne) hrskavice unutar tela pršljena pri čemu je razviće kičmenih pršljenova roda Triturus veoma slično razviću kod drugih vrsta repatih vodozemaca. Homeotske transformacije predstavljaju transformacije pri kojima jedan deo tela preuzima identitet drugog dela tela (npr. transformacija trupnog u sakralni pršljen) usled mutacije ili promena u ekspresiji određenih Hox gena. Studija variabilnosti u broju trupnih pršljenova i tipova homeotskih transformacija urađena je na uzorku koji pokriva celokopnu genetičku i geografsku varijabilnost roda Triturus. Uočeno je odsustvo korelacije između nepotpunih homeotskih transformacija sakralnog pršljena i varijabilnosti u broju trupnih pršljenova što može biti rezultat razvojnih mehanizama koji favorizuju kompletan broj presakralnih pršljenova ili favorizovanje selekcije protiv tranzicionih pršljenova kod ove grupe repatih vodozemaca. Takođe, u ovoj tezi izučavane su i evolucione promene u broju, veličini i obliku trupnih pršljenova kod adultnih jedinki devet vrsta roda Triturus. Metodama kompjuterske mikrotomografije i 3D geometrijske morfometrije uz konstruisanje filomorfološkog prostora analizirane su promene u obliku i veličini samih kičmenih pršljenova odnosno procenjeno je u kojoj meri su evolucione promene u obliku korelisane sa promenama u veličini i broju kičmenih pršljenova. Ovi rezultati su pokazali da se vrste roda Triturus razlikuju u broju, veličini i obliku trupnih pršljenova. Elongacija tela, koja je u vezi sa akvatičnijim načinom života, postignuta je povećanjem broja kičmenih pršljenova, a interspecijske razlike u obliku trupnih pršljenova korelisane su sa elongacijom. Species- specifične razlike u obliku trupnih pršljenova su takve da se vrste mogu sa sigurnošću identifikovati i na osnovu samo jednog pršljena, što omogućava eventualnu primenu ovih metoda u identifikaciji fosilnog i subfosilnog materijala.

Analyses of intra- and interspecific morphological variation through ontogeny (e.g. difference in the time of differentiation, rate and direction of ontogenetic changes in size and shape), as well as variability at the adult stage, enable determination of morphological patterns and mechanism which lead to evolution of morphological structures. In this thesis, morphological variability of axial skeleton was studied within monophyletic group of crested newts which comprises nine species of the genus Triturus. Crested newts are suitable model organisms for studies of vertebral development and evolution. They are closely related group of species with known phylogenetic relations, different rate of body elongation and great variability in the number of trunk vertebrae. Histological techiques were used in tracing postembrionic development of trunk verteberae to test difference in the time of vertebral development, ossification sequences and rate of vertebral differentiation. For these analyses, two species of crested newts which differ in rate of body elongation and number of trunk vertebrae were chosen. Results show that there is no difference between analysed species of Triturus newts in development time and formation of vertebrae. Three modes of the ossification process were recorded during vertebral formation: 1) perichordal ossification, 2) perichondrial ossification and 3) endochondral ossification. Also, the presence of intravertebral (notochordal) cartilage was recorded at centrum which makes vertebral development in Triturus newts very similar to other species of tailed amphibians. Homeotic transformations are replacements of one body part by another (e.g. transformation of trunk into sacral vertebra) due to the mutation or changes in expression of certain Hox genes. The studies of variability in the number of trunk vertebrae and types of homeotic transformations were done on sample that represents all genetical and geographical variability in the genus Triturus. The absence of correlation between the frequency of incomplete homeotic transformation of sacral vertebrae and variation in the number of trunk vertebrae could be a result of developmental mechanisms that favour complete numbers of presacral vertebrae or selection against transitional vertebrae in this group of tailed amphibians. Also, in this thesis, evolutionary changes in number, size and shape of trunk vertebrae of adult specimens were studied across nine species of Triturus newts. The employed methods of computed microtomography and three-dimensional geometric morphometrics with construction of phylomorphospace enabled us to analyze changes in shape and size of vertebrae, i.e. to which extent evolutionary changes in vertebral size are correlated with changes in vertebral shape. These results show significant difference in number, size and shape of trunk vertebrae among Triturus species. Body elongation, which is related to more aquatic lifestyle, was achieved through an increase in the number of trunk vertebrae, and interspecific differences in vertebral shape are correlated with this pattern of elongation. Species-specific differences in vertebral shape are such that single trunk vertebra can be used for the identification of species, which enables eventual use of these methods for identification of fossil and subfossil material.

Analiza polimorfizama gena za receptor za vitamin D i gena asociranih sa trombofilijom kod žena sa idiopatskim infertilitetom

Analysis of the polymorphisms in vitamin D receptor gene and genes associated with thrombophilia in women with idiopathic infertility

Uvod. Genetičke analize mogu ukazati na uzrok infertiliteta kod ţena kod kojih klinički testovi nisu uspeli da utvrde razloge reproduktivnog neuspeha. Cilj. Cilj ovog rada je da se ispita postojanje veze genskih polimorfizama asociranih sa trombofilijom i polimorfizama u genu za receptor za vitamin D (VDR) sa idiopatskim infertilitetom. Materijal i metode. U studiju je uključeno 117 pacijentkinja sa idiopatskim infertilitetom, kao i 130 ţena sa najmanje jednom trudnoćom realizovanom bez komplikacija. Primenom TaqMan metode uzorci su genotipizirani u odnosu na polimorfizme: FV 1691 G>A, FII 20210 G>A, MTHFR 677 C>T, MTHFR 1298 A>C, PAI-1 -675 4G/5G, ATIII 786 G>A, ACE I/D i ITGB3 1565 T>C, dok su polimorfizmi u VDR genu analizirani primenom restrikcionih enzima (FokI, BsmI, ApaI i TaqI). Rezultati. Ispitivanjem etioloških faktora pokazano je da je porodična anamneza značajan faktor pri proceni individualnog rizika za infertilitet. Utvrđeno je da visokorizične varijante FV 1691A i FII 20210A predstavljaju nezavisne faktore rizika za nastanak primarnog infertiliteta. Analizom multilokusnih interakcija definisani su kompleksniji genotipovi asocirani sa sekundarnim infertilitetom. Ispitivanja polimorfizama u VDR genu pokazala su postojanje protektivne uloge alela F u FokI i alela B u BsmI polimorfizmu. Pored toga, analizom vezanosti markera u VDR genu identifikovano je postojanje haplotipova, od kojih je bAT povećavao rizik za sekundarni infertilitet, dok je BAT imao protektivnu ulogu za nastanak primarnog infertiliteta. Zaključak. Analiza polimorfizama u genima asociranim sa trombofilijom i genu za VDR ne samo da definiše moguće kliničko-genetičke dijagnostičke procedure, već sugeriše moguće mehanizme za odrţavanje hemostatskog i imunološkog balansa u procesu reprodukcije.

Introduction. Genetic analysis may indicate cause of infertility in women to whom clinical trials have failed to determine the causes of reproductive failure. Aim. The aim of this study is to investigate a link between polymorphisms associated with thrombophilia and polymorphisms in the vitamin D receptor gene (VDR) with idiopathic infertility. Material and methods. This study included 117 female patients with idiopathic infertility, as well as 130 women with at least one pregnancy without complications. Using TaqMan method, the samples were genotyped for polymorphisms: FV 1691 G>A, FII 20210 G>A, MTHFR 677 C>T, MTHFR 1298 A>C, PAI-1 -675 4G/5G, ATIII 786 G>A, ACE I/D and ITGB3 1565 T>C, while VDR gene polymorphisms were analyzed using restriction enzymes (FokI, BsmI, ApaI and TaqI). Results. Examination of etiological factors has shown that family history is a significant factor in assessing individual risk for infertility. High risk variants FV 1691A and FII 20210A have been shown to be independent risk factors for occurrences of primary infertility. An analysis of multilocus interactions defined more complex genotypes associated with secondary infertility. Polymorphisms testing in VDR gene showed the existence of a protective role of allele F in FokI and allele B in BsmI polymorphisms. In addition, the analysis of the linkage in VDR gene identified the existence of haplotypes, of which bAT increased the risk of secondary infertility, while BAT had a protective role of primary infertility. Conclusions. The analysis of polymorphisms in the genes associated with thrombophilia and the VDR gene not only defines possible clinical-genetic diagnostic procedures, but suggests possible mechanisms for maintaining hemostatic and immune balance in the reproductive process.