Neuroglobin and cytoglobin distribution patterns in human and canine eye

Globins are a family of heme-containing proteins that reversibly bind gaseous ligands such as oxygen, nitric oxide and carbon monoxide. Neuroglobin (Ngb) and cytoglobin (Cygb) have been recently added to mammalian globin family. The general hypothesis of this dissertation was: The presence of Ngb and Cygb can be detected by biochemical and immunohistochemical methods in different structures of the eye;Ngb immunoreactivity (IR) in the canine retina was located in the ganglion cell layer (GCL), inner (INL) and outer nuclear layers (ONL), inner (IPL) and outer plexiform layers (OPL), photoreceptor inner segments (IS) and retinal pigment epithelium (RPE). Cygb-IR in the canine retina was found in the GCL, INL and ONL, IPL and OPL and RPE. Ngb and Cygb were expressed in the same cells in the GCL and INL. Distribution pattern of Ngb and Cygb in human retina was similar to distribution found in the canine retina;Ngb and Cygb-IR in the human and canine anterior segment structures was detected in the corneal epithelium and endothelium. Furthermore, in the iris, Ngb and Cygb-IR was localized to the anterior border layer and the stroma, iridal sphincter and dilator muscle. In the iridocorneal angle, both Ngb and Cygb were detected in endothelial cells of the human and canine trabecular meshwork and canal of Schlemm in human. In the ciliary body, Ngb- and Cygb-IR was localized to the nonpigmented ciliary epithelium of the pars plana and pars plicata, as well as in ciliary body musculature. Weak Ngb and Cygb-IR was detected in the lens epithelium. Ngb and Cygb distribution was consistent between human and canine anterior segments and was co-localized within the same cells in all structures;Our studies are the first detailed description of Ngb and Cygb presence detected by immunohistochemical methods in different structures of the canine and human eye. Based on Ngb and Cygb localization and their previously reported biochemical features, we hypothesize that Ngb and Cygb may have important roles in scavenging reactive oxygen species and/or facilitating oxygen diffusion in the eye

Optimization and validation of HPLC method for the determination of amino acids

Cilj ovog istraživanja bio je optimizirati i validirati metodu za odreĎivanje asparagina, glutaminske kiseline, serina, arginina, prolina, treonina, metionina i izoleucina pomoću tekućinske kromatografije visoke djelotvornosti (HPLC). Optimizacija metode obuhvaćala je optimizaciju uvjeta derivatizacije aminokiselina te uvjeta kromatografske analize. Validacija metode provedena je uz derivatizaciju u trajanju od 20 minuta, pri utvrĎenom optimalnom gradijentnom eluiranju te pri temperaturi kolone od 50 °C. Ključni parametri validacije bili su selektivnost, linearnost i radno područje, preciznost, granica detekcije, granica kvantifikacije i robusnost. Budući da su rezultati provedenih validacijskih ispitivanja u skladu s postavljenim kriterijima prihvatljivosti, može se zaključiti da je metoda selektivna, linearna unutar radnog područja te precizna u odreĎivanju ciljanih analita. UtvrĎena je stabilnost derivata aminokiselina u vremenu od dva sata pri +4 °C.The aim of this study was to optimize and validate a method for determination of asparagine, glutamic acid, serine, arginine, proline, threonine, methionine, and isoleucine using high performance liquid chromatography (HPLC). Method optimization included optimization of derivatization conditions and conditions of chromatographic analysis. Method was validated under optimal conditions including 20 minute derivatization time, optimal gradient elution and column tempereture of 50 °C. Key validation parameters were selectivity, linearity and working range, precision, limit of detection, limit of quantification, and robustness. The results of the validation experiments were in accordance with defined criteria and can be concluded that the method is selective, linear under working range and precise in determination of target analytes. Stability of the amino acid derivatives was determined for two hours at +4 °C

Prognostičko značenje izraženosti proteina NEDD9 u transbronhalnim bioptatima u bolesnika s karcinomom pluća [Prognostic significance of NEDD9 in small biopsies of lung cancer]

Lung cancer is the major cause of cancer mortality worldwide and includes several histologic subtypes evolving from numerous genetic and epigenetic changes emerging in alveolar, bronciolar and bronchial epithelium. Precise histologic diagnosis is the cornerstone of the individualised treatment of lung cancer. Here we have demonstrated significantly higher accuracy and reproducibility of the diagnosis using immunohistochemistry in transbronchial biopsies of lung adenocarcinoma (p=0,005). Also, only 5.41% of all samples was finally diagnosed as NSCC NOS. Positivity for napsin A was seen in 45.33% of all adenocarcinoma samples along with consistant coexpression of TTF-1 in 90% of cases (p=0,289). As for CK7, homogenous immunoreactivity showed in 91.89% of all adenocarcinomas, and therefore staining for napsin A wasn't proven superior to neither CK 7 nor TTF-1 in adenocarcinoma immunotyping. Nevertheless, napsin A was negative in 95% of squamous cell carcinoma samples and consequently may be useful as an additional marker of glandular differentiation in less clear immunoprofiles (p<0,001). Regulatory protein NEDD9 is involved in control of migration chemotaxis, apoptosis, cell cycle and cellular differentiation, playing the role of the hub-protein in the complex process of epithelial-mesenchimal transition. NEDD9 has recently been identified as integral part of aquisition of metastatic potential in mammary adenocarcinoma, glioblastoma and metastatic melanoma. We have demostrated both nuclear and cytoplasmatic expression of NEDD9 in transbronchial biopsies of lung adenocarcinoma, squamous carcinoma and also small cell carcinoma indicating the activation of the epithelial-mesenchimal transition in different lung cancer subtypes. Cytoplasmatic expression of NEDD9 was higher in comparison to nuclear in all carcinoma subtypes (p<0.001). The direct relationship between the expression of NEDD9 and survival wasn't established

Problemi konkurentnosti privrede Srbije u uslovima evropskih integracija

Konkurentnost nacionalnih privreda se nalazi u fokusu svih studija koje su posvećene analizi položaja zemalja u svetskoj ekonomiji. U savremenim uslovima privređivanja zemlje se neprekidno nadmeću za oskudne resurse, te se njihova sposobnost da privuku strane investicije i koriste ova ograničena redstva posmatra kao njihova glavna konkurentska prednost. Nepovoljan poslovni ambijent, pre svega, u zemaljama u razvoju, smanjuje priliv inostranih sredstava i izvoz, što neminovno dovodi do smanjenja životnog standarda stanovništva i usporavanja ekonomskog rasta. Jačanje konkurentnosti privrede Srbije jedan je od primarnih zadataka kako bi se sprečilo dalje produbljivanje razvojnog jaza između Srbije i drugih evropskih zemalja i, s tim u vezi i zaustavio trend daljeg povećanja nesrazmere u razvoju pojedinih regiona unutar same zemlje. Proevropski razvoj Srbije i njeno učlanjenje u EU predstavlja razvojnu šansu za Srbiju, ali i najefikasniji garant stabilnog demokratskog razvoja, ujednačenog ekonomskog napredovanja, kao i garancija unutrašnje i spoljne bezbednosti građana i države. Prioritetni tretman koji razvoju održivih i konkurentskih privreda daje EU je polazište za sagledanje značaja ove oblasti za proces približavanja Srbije EU i istovremeno jedna od repernih tačaka za tumačenje trenutnog stanja razvoja konkurentsnosti u Srbiji