The human SOX18 gene: Expression analysis and characterization of its 5’ flanking region

The aim of this study was to establish an adequate in vitro model system for studying transcriptional regulation of the human SOX18 gene. The paper presents an analysis of expression of this gene in cultured cell lines and characterization of its 5' flanking region. Using RT-PCR, Northern and Western blot analysis, we demonstrated SOX18 expression in HeLa cells, indicating that this cell line provides a suitable model system for studying transcriptional regulation of the given gene. We also cloned, sequenced and for the first time characterized the human SOX18 5’ flanking region. It is shown that the region 892 bp in size immediately upstream from the start codone harbors regulatory elements sufficient for transcription and represents an SOX18 promoter region

PCR amplification and sequence analysis of mole rat (Spalax leucodon) Sox2 gene

Sox2 gen je uključen u kontrolu razvića oka i kod sisara je pokazano da je transkripcioni aktivator γF-kristalina u sočivu oka. Slepo kuče (Spalax leucodon), je ekstremni primer prirodne degeneracije oka. Ova životinja nema spoljašnje oči, a sočivo je neorganizovano i vakuolizovano. U ovom radu smo prikazali parcijalnu karakterizaciju Sox2 gena slepog kučeta. Gen je kloniran korišćenjem pristupa zasnovanog na PCR tehnologiji (upotrebom 'prajmera' specifičnih za SOX2 gen čoveka). Naši rezultati ukazuju da u otvorenom okviru čitanja Sox2 gena kod slepog kučeta nema značajnih promena u odnosu na do sada okarakterisane Sox2 gene kod sisara. Zapaža se snažna konzervisanost sekvence u poređenju sa ortolognim genima čoveka, ovce i miša. Naši rezultati ukazuju da je Sox2 gen slepog kučeta pod selektivnim pritiskom, što ukazuje da sem u razviću oka verovatno ima i druge važne funkcije u embriogenezi.Sox2 gene is involved in control of eye development and it has been shown that in mammals this gene is a transcriptional activator of γF-crystallin in lens. The mole rat (Spalax leucodon), which is an extreme example of natural visual degeneration, has strongly rudimented eyes with undeveloped, disorganized and vacuolated lens nuclei. Here we present a partial characterization of the mole rat Sox2 gene which is cloned using PCR - based approach (and primers specific for human SOX2 gene). Our results have shown no significant changes in the sequence within the open reading frame of the mole rat Sox2 gene and have revealed that mole rat Sox2 gene is strongly conserved compared with orthologue gene in humans, sheep and mouse. Our results indicate that the mole rat Sox2 gene remains under selective pressure since it probably has, apart from the control of eye development, the other important roles in the embryonic development

Zaposleni u zdravstvu u transformaciji zdravstvenog menadžmenta

Introduction: The importance of studying management in one of the most sensitive and complex areas of human endeavor that is healthcare is of great significance for providing high quality and efficient easily-accessible healthcare in the future. Studying the challenges that need to be overcome towards achieving this goal is of great importance not only from the viewpoint of scientific management theory but primarily to help bring about strategically important decisions that are crucial for the continued evolution and reformation of the healthcare system, in which the employees have a new, much more complex role. The objective: was to more thoroughly study, describe and understand the role the employees have in the transformation of health care management. Material and method: During this research the material used was foreign and local literature, processed using the Analytical method as well as the Observation and Comparative methods. Conclusion: In the transformation of the health care management a very important role is played by the health care employees. Without the transformation of the current practice and significant changes in both the structure and work ethic of all the employees in health care services (nurses and people in management alike) there will be no improvement in the health care system overall.Uvod: Značaj proučavanja menadžmenta u jednom od najkompleksnijih i najosetljivijih ljudskih delatnosti kao što je zdravstvo od izuzetne je važnosti za budućnost pružanja kvalitetne i efikasne, svakom dostupne zdravstvene zaštite. Važnost proučavanja ove problematike bitna je ne samo sa gledišta naučne teorije o menadžmentu, već, pre svega, radi donošenja strateški važnih odluka za dalje funkcionisanje i razvoj novog, reformisanog zdravstvenog sistema u kome zaposleni u zdravstvu imaju novu, mnogo kompleksniju ulogu. Cilj rada je bio da se jasnije prouči, opiše i sagleda uloga zaposlenih u zdravstvu u transformaciji zdravstvenog menadžmenta. Materijal i metod: Prilikom istraživanja ove teme materijal za rad je bila strana i domaća literatura, obrađena korišćenjem Analitičkih metoda, a koristio se i Metod posmatranja , kao i Komparativne metode. Zaključak: Uloga zaposlenih u zdravstvu u transformaciji zdravstvenog menadžmenta je izuzetno kompleksna i važna. Bez temeljne transformacije dosadašnje prakse rukovođenja u zdravstvu i bez značajnih promena, kako u strukturi, tako i u radu svih zaposlenih u zdravstvenim službama, bilo da se radi o medicinskom ili ne medicinskom osoblju, neće biti ni poboljšanja u zdravstvenom sistemu u celini

The story of the SOX genes: for better or for worse…

The SOX genes encode a group of transcription factors showing diverse expression patterns during de- velopment and acting as key regulators of diverse cellular processes. SOX transcription factors are in- volved in multiple events from the maintaining of stem cells pluripotency and cell fate decision to driving terminal differentiation of cells into specialized cell types. During adulthood SOX transcription factors control various physiological processes. Mutations in SOX genes have been associated with severe clin- ical disorders, while deregulation of their expressions cause a broad range of pathological condition. Ac- cumulating evidence suggests that SOX proteins act as oncogenes and recent evidence points toward pro-proliferative, pro-survival and/or anti-differentiation roles of the SOX proteins. The results of long-term research of the structure, regulation of expression and the function of selected SOX genes will be presented. It will include data obtained by studying the roles of SOX genes in in vitro neural differentiation of pluripotent embryonal carcinoma cells, as well as interaction of SOX transcrip- tion factors with signalling pathways active during neurogenesis and oncogenesis. Special focus will be made on ongoing research focused on the roles of SOX genes in promotion of malignant phenotype of cancer cells and maintaining of cancer stem cells. The interplay of SOX transcription factors and mi- croRNAs in the brain under physiological and pathological conditions, along with crosstalk between SOX genes and long non-coding RNAs in glioblastoma will be discussed. The role of SOX transcription factors in ageing and age-related diseases will be outlined

Some experiences in tapping deep thermal waters of the Triassic karstic aquifer in the Pannonian Basin of Serbia

The Triassic karstic aquifer is the system with the greatest potential for the utilization of thermal waters in Serbia. As an integral part of the Dinaric tectonic unit, the Triassic aquifer extends widely over the western part of the Serbian territory and is characterized by cold waters. In contrast, the same but confined type of aquifer overlain by thick Tertiary sediments in the Pannonian Basin has significant geothermal potential. The major potential for tapping geothermal flow is in the southern and southwestern parts of the Pannonian Basin (Srem) and in the adjacent areas of Mačva and Semberija in the Sava tectonic graben. In these areas the Triassic karstic aquifer has been tapped by several boreholes with depths ranging from 400 m to 2400 m. The temperature of the hottest water exceeds 75 °C, while maximal discharge is 40 l/s. Although the prospect of wider utilization of geothermal energy undoubtedly exists, some Serbian national plans count on a limited contribution of geothermal energy in renewable energy sources of only 4%. This is probably due to the low level of current utilization, and the inefficient use of even some highly productive wells with a high water temperature, such as those drilled in the most prosperous Mačva region

Retinoic acid-induced SoX3 gene expression in NT2/D1 cells is RXR homodimer-independent

Sox3/SOX3 gen je uključen u kontrolu razvića nervnog sistema. Naša prethodna istraživanja su pokazala da tokom neuralne indukcije NT2/D1 ćelija retinoičnom kiselinom (RK) dolazi do promene ekspresije humanog SOX3 gena. Takođe, pokazali smo da su RXR retinoidni receptori glavni posrednici efekta RK na SOX3 ekspresiju, dok su članovi RAR familije receptora isključeni kao njihovi heterodimerni partneri u ovom signalnom putu. Rezultati predstavljeni u ovom radu ukazuju da je aktivacija SOX3 gena retinoičnom kiselinom nezavisna od RXR homodimera. Ova izučavanja ekspresije SOX3 gena su značajna za buduća istraživanja uticaja koji ovaj gen ima na različite aspekte normalnog i patološkog razvića.The Sox3/SOX3 gene is implicated in the control of nervous system development. We previously demon­strated modulation of human SOX3 gene expression during neural induction of NT2/D1 cells by retinoic acid (RA). Also, we accurately verified RXR retinoid receptors as major mediators of the effect of RA on SOX3 expression, and excluded RARs as its heterodimeric partners in RA-SOX3 signaling. Here we present evidence that activation of the SOX3 gene by RA is not RXR homodimer-dependent. The described line of SOX3 gene expression studies is valuable for future investigation of the impact that this gene has multiple aspects of normal and pathological development

Gene expression analysis by non-radioactive RNA-RNA in situ hybridization techniques

RNK-RNK in situ hibridizacija je pogodna metoda za izučavanje specifične ekspresije gena u tkivima i ćelijama, kojom je omogućena vizualizacija obeležene antisens RNK probe hibridizovane za specifičnu iRNK. U ovom radu prikazano je detektovanje ekspresije SOX gena u tumorskim ćelijskim linijama primenom neradioaktivne RNK-RNK in situ hibridizacije koristeći RNK probe obeležene biotinom ili digoksigeninom. Koristeći ovaj pristup potvrđeni su rezultati dobijeni Northern blot analizom, tj. pokazano je prisustvo SOX2 iRNK u NT2/D1 i SOX14 iRNK u HepG2 ćelijama. Cilj ovog rada bio je uspostavljanje RNK-RNK in situ hibridizacije na in vitro kultivisanim ćelijama da bi metodu primenili pri proučavanju ekspresije SOX gena u različitim normalnim i tumorskim tkivima.RNA-RNA in situ hybridization is a reliable method for studying tissue and cell specific gene expression, which enables visualization of labeled antisense RNA probe hybridized to specific mRNA. In this study we employed non-radioactive RNA-RNA in situ hybridization using biotin- or digoxigenin-labeled RNA probes in order to detect SOX gene expression in carcinoma cell lines. By this approach we confirmed results obtained by Northern blot analysis, where the presence of SOX2 mRNA in NT2/D1 and SOX14 mRNA in HepG2 cells has been established. Our aim was to set up RNA-RNA in situ hybridization method in in vitro cultured cells in order to perform further analyses of SOX gene expression on various normal and cancer tissues

Establishment and initial characterization of SOX2-overexpressing NT2/D1 cell clones

SOX2, a universal marker of pluripotent stem cells, is a transcription factor that helps control embryonic development in vertebrates; its expression persists in neural stem/progenitor cells into adulthood. Considering the critical role of the SOX2 transcription factor in the regulation of genes required for self-renewal and pluripotency of stem cells, we developed and characterized SOX2-overexpressing NT2/D1 cell clones. Using Southern blot and semi-quantitative RT-PCR, we confirmed integration and expression of exogenous SOX2 in three NT2/D1 cell clones. Overexpression of the SOX2 gene was detected in two of these clones. SOX2 overexpression in NT2/D1 cell clones resulted in altered expression of key pluripotency genes OCT4 and NANOG. Furthermore, SOX2-overexpressing NT2/D1 cell clones entered into retinoic acid-dependent neural differentiation, even when there was elevated SOX2 expression. After 21 days of induction by retinoic acid, expression of neural markers (neuroD1 and synaptophysin) was higher in induced cell clones than in induced parental cells. The cell clone with SOX2 overexpression had an approximately 1.3-fold higher growth rate compared to parental cells. SOX2 overexpression did not increase the population of cells undergoing apoptosis. Taken together, we developed two SOX2-overexpressing cell clones, with constitutive SOX2 expression after three weeks of retinoic acid treatment. SOX2 overexpression resulted in altered expression of pluripotency-related genes, increased proliferation, and altered expression of neural markers after three weeks of retinoic acid treatment

New directions in the design of railways stations

Održivi razvoj prometa i gradova, objedinjavanje vidova transporta, preporod željezničkog prometa, utjecali su na izgradnju velikog broja željezničkih kolodvora posljednjih desetljeća. Multimodalni kolodvori, unaprijeđenih koncepata i integrirani u urbanu matricu grada, postaju novi centri razvoja. U radu se iz aspekta urbanističko-arhitektonske struke analiziraju pozitivna iskustva u izgradnji značajnih željezničkih kolodvora. Rezultati provedenih analiza na primjeru željezničkog kolodvora u Beogradu, prema predloženom morfološkom konceptu, mogu poslužiti kao smjernice u definiranju važnih kolodvora u urbanim sredinama.Sustainable development of traffic and cities, integration of transport modes, and the renaissance of railway traffic, have led to construction of a great number of railway stations/terminals over the past decades. These multi-modal traffic interchanges, based on advanced concepts and properly integrated in urban matrices, are becoming new focal points of urban development. Recent positive experience in the construction of significant railway stations is presented from the standpoint of urban development. Results obtained during analysis of the Belgrade railway station, which is based on the proposed morphological concept, can be used as guidelines for development of other significant railway stations in urban areas