Low dose gamma-irradiation alters the expression of p53 protein in the rat hippocampus

The tumor-suppressor p53 protein and glucocorticoid receptor (GR) respond to different types of stress. In an attempt to reveal the possibility that p53 protein is involved in the regulation of GR gene expression after low dose cranial irradiation (CI) with 2 Gy, we examined the expression of p53 mRNA and protein as well as expression of GR mRNA and protein in the hippocampus of 18-days-old rats. We found that p53 mRNA expression was unchanged after CI, while induction of p53 protein was rapid, leading to the accumulation of p53 protein in the cytoplasm. Irradiation leads to stimulated GR gene expression in a time-dependent manner, whereas the level of GR protein was unchanged after CI. Co-immunoprecipitation has not showed that wild type p53 protein physically interacts with the GR in the cytoplasm. Our data suggest that the low dose cranial irradiation leads to stabilization of the 53 protein, without interaction with GR protein the cytoplasm.Physical chemistry 2006 : 8th international conference on fundamental and applied aspects of physical chemistry; Belgrade (Serbia); 26-29 September 200

Different induction of dual corticosteroid receptor system in the rat hippocampus following gamma radiation

Cranial radiotherapy (CRT) is an effective way to prevent CNS relapse in children with acute lymphoblastic leukemia (ALL). However, CRT also has serious side effects on normal tissues, including long-term neuroendocrine disturbances. In order to test this clinical protocol on animals, we examined the effects of CRT (10 Gy) on the level of mRNA for glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) in the hippocampus of 8-days-old rats. Irradiation rapidly stimulated GR gene expression in a time-dependent manner, whereas the time-course of MR mRNA expression showed no statistically significant changes. At postnatal day 42, the level of GR mRNA was diminished while the level of MR mRNA remained unchanged compared to matched controls. Dexamethasone suppression test (DST) revealed the altered nucleocytoplasmic shuttling of activated GR after CRT in 42-days-old rats, as a long-term consequence of gamma irradiation.Physical chemistry 2006 : 8th international conference on fundamental and applied aspects of physical chemistry; Belgrade (Serbia); 26-29 September 200

The effect of lithium on the neurotransmitter and antioxidant status in the perfrontal cortex and hippocamus of the rats exposed to restraint stress

Cilj istraživanja je bio da se detaljnije prouče molekulski mehanizmi delovanja litijuma na neurotransmiterski i antioksidativni status u prefrontalnom korteksu i hipokampusu stresiranih životinja. Specifični cilj rada bio je da se ispita uticaj ponavljanog stresa fizičkog sputavanja u trajanju od 14 dana, kao i delovanje litijuma kod pacova izlaganih 14 dana ponavljanom stresu fizičkog sputavanja na: količinu dopamina (DA) i noradrenalina (NA), ekspresiju gena za tirozin hidroksilazu (TH) i dopamin β hidroksilazu (DBH) (enzime koji učestvuju u biosintezi DA i NA), ekspresiju gena za dopaminske i noradrenalinske transportere odgovorne za preuzimanje, skladištenje i odpuštanje DA i NA (DAT, NET i VMAT 2), aktivnost i količinu enzima odgovornih za razgradnju DA i NA (MAO A, MAO B i COMT), ekspresiju gena za enzime antioksidativne zaštite (SOD 1, SOD 2, CAT, GPx i GR), ekspresiju gena za BDNF u prefrontalnom korteksu i hipokampusu, koncentraciju ACTH i CORT u plazmi i ponašanje eksperimentalnih životinja. U eksperimentima je korišćen ponavljani stres fizičkog sputavanja, izvođen na taj način što je pacov podvrgnut stanju nepokretnosti smeštanjem u plastične tube dimenzija 25x7 cm. Pacovi su podvrgnuti opisanom stresu po dva sata tokom 14 dana u različito vreme u periodu od 9-13 h. Rastvor litijum-hlorida (LiCl) je pacovima injeciran intraperitonealno jednom dnevno 14 dana. Početna doza litijuma je bila 1,5 mEq/kg i primenjena je prva dva dana. Zatim je povećana na 2,3 mEq/kg sedam dana, pa na 3 mEq/kg pet dana. Ovakav protokol davanja LiCl pacovima obezbeđuje koncentracije litijuma u plazmi koje su, u poređenju sa pacijentima sa bipolarnim poremećajem, iznad minimalne terapeutske koncentracije (i. e. 0,4 mM) tokom perioda terapije. U ovom istraživanju korišćene su sledeće metode: 1. Metoda za kvantifikaciju ekspresije TH, DBH, DAT, NET, VMAT 2, SOD 1, SOD 2, CAT, GPx, GR i BDNF gena (RТ-PCR-а u realnom vremenu sa TaqMan probama); 2. Metoda za kvantifikaciju sinteze TH, DBH, DAT, NET, VMAT 2, COMT, SOD 1, SOD 2, CAT, GPx, GR i BDNF proteina (Wеstern blot); 3. Metode za određivanje aktivnosti enzima MAO A i MAO B (Amplex Red Monoamine Oxidase Assay kit; molecular Probes), SOD 1, SOD 2, CAT, GPx i GR (Randox Laboratories, Crumlin, UK; metoda po Claiborn-u; metoda po Maral-u; metoda po Glatzle-u); 4. Metoda za određivanje DA i NA u tkivu (kit 3-CAT Research ELISA kits BA E-5600, Labor Diagnostica Nord, Nordhorn, Germany); 5. ELISA еsеј, metoda za određivanje CORT u plazmi (OCTEIA EIA kit, Immunodiagnostic Systems Inc.); 6 Hemiluminiscentna metoda za određivanje ACTH u plazmi pomoću automatskog analizatora...The aim of this research was to investigate the molecular mechanisms underlying the effects of lithium on neurotransmitter and antioxidant status in prefrontal cortex and hippocampus of the stressed animals. The specific aim of the work was to investigate the effect of chronic restraint stress in a period of 14 days and also the effect of lithium administration on: the amounts of dopamine (DA) and noradrenaline (NA), the gene expression of tyrosine hydroxylase (TH) and dopamine-β-hydroxylase (DBH) (biosynthetic enzymes for DA and NA), the gene expression of transport proteins for uptake, storage and release of DA and NA (DAT, NET and VMAT 2), the activity and amount of DA and NA degrading enzymes (MAO A, MAO B and COMT), the gene expression of antioxidant enzymes (SOD 1, SOD 2, CAT, GPx and GR), the gene expression of BDNF in prefrontal cortex and hippocampus, the plasma concentrations of ACTH and CORT, as well as on the behavior of experimental animals. In our experiments the rats were subjected to chronic restraint stress by being placed in the plastic tubes sized 25x7 cm, two hours a day for 14 days, at different times in the period from 9 to 13 h. Lithium-chloride solution (LiCl) was injected to the animals intraperitoneally once a day during 14 days. The initial dose of lithium of 1,5m Eq/kg was applied in the first two days. The dose was increased to 2,3 mEq/kg during the next 7 days and to 3 mEq/kg during the following 5 days. This protocol of LiCl administration provided the concentrations of lithium in the plasma of rats to be higher than the minimal therapeutic concentration (i.e. 0,4 mM) in the patients with bipolar disorder. The following methods were used in this investigation: 1. The expressions of TH, DBH, DAT, NET, VMAT 2, SOD 1, SOD 2, CAT, GPx, GR and BDNF were quantified by using real time RT-PCR with TaqMan assays. 2. Western blot analysis was used to quantify the synthesis of TH, DBH, DAT, NET, VMAT 2, COMT, SOD1, SOD 2, CAT, GPx, GR and BDNF protein. 3. Methods for evaluation of MAO A and MAO B enzyme activity (Amplex Red Monoamine Oxidase Assay kit; molecular Probes), enzyme activities of SOD 1, SOD 2, CAT, GPx i GR (Randox Laboratories, Crumlin, UK; method by Claiborn-u; method by Maral-u; method by Glatzle-u); 4. Methods for measuring DA i NA in the tissue (kit 3-CAT Research ELISA kits BA E-5600, Labor Diagnostica Nord, Nordhorn, Germany); 5. ELISA assay, the method for measuring CORT in the plasma (OCTEIA EIA kit, Immunodiagnostic Systems Inc.); 6. Hemiluminiscent method for measuring of ACTH in the plasma by automatic analyzer..

Unmet health care needs as an indicator of availability, accessibility and acceptability of health care

Uvod: Nezadovoljene potrebe za zdravstvenim uslugama su prepoznate kao pokazatelj pristupačnosti zdravstvene zaštite, koji daje sliku o specifičnim preprekama sa kojima se korisnici suočavaju kada imaju potrebu za korišćenjem zdravstvenih usluga. Cilj ove disertacije je bio da analizira demografske, socijalnoekonomske, regionalne karakteristike i percepciju zdravstvenog stanja ispitanika koji imaju nezadovoljene zdravstvene potrebe za uslugama lekara i uslugama stomatologa u 2013. i 2014. godini i da identifikuje prediktore za nastanak nezadovoljenih potreba za uslugama lekara i uslugama stomatologa koji se odnose na dostupnost, pristupačnost i prihvatljivost zdravstvene zaštite. Metod: U ovoj studiji korišćeni su podaci iz dva nacionalna istraživanja o prihodima i uslovima života (Anketa o prihodima i uslovima života - SILC), sprovedenih u Srbiji na reprezentativnom uzorku osoba starijih od 15 godina (17 183) u 2013. godini i starijih od 16 godina (16 220) u 2014. godini. .Kao instrument ispitivanja korišćeni su upitnici. Rezultati: U 2013. godini skoro svaki peti ispitanik stariji od 16 godina je imao nezadovoljene potrebe za uslugama lekara, a u 2014. godini skoro svaki sedmi ispitanik. Razlozi nezadovoljenih potreba za uslugama lekara koji ukazuju na odgovornost sistema zdravstvene zaštite, (ugrožena pristupačnost i dostupnost zdravstvene zaštite), su u 2013. godini bili odgovorni za 44,4% nezadovoljenih potreba a u 2014. godini za 58,2%. Razlozi koji su bili u vezi sa ličnim preferencijama ispitanika u 2013. godini su bili zastupljeni u 55,6% slučajeva a u 2014. godini u 41,7% slučajeva. Najdominantniji razlog u okviru ugrožene pristupačnosti je bio finansijski razlog, u okviru ugrožene dostupnosti to su bile liste čekanja na medicinske intervencije i procedure a u okviru ugrožene prihvatljivosti želja da se sačeka i vidi da li će se stanje zdravlja popraviti i problem proći sam od sebe...Introduction: Unmet health care needs are recognized as an indicator of access to health care, which provides an overview of specific barriers that users face when they need health services. This thesis aimed to analyze demographic, socioeconomic, regional characteristics and perception of the health status; to identify predictors of unmet health care needs and consequently determine the size of inequalities in the availability, accessibility and acceptability of health care. Method: The study used data from two national surveys on Income and living conditions (Survey on Income and Living Conditions - SILC), conducted in Serbia on a representative sample of people over 15 years old (17,183) in 2013 and older than 16 years (16,220) in 2014. The survey instruments were questionnaires. Results: In 2013 every fifth respondent over the age of 16 had unmet health care needs while in 2014 every seventh respondent. The reasons for unmet health care needs related to the availability of the health care, (threatened availability and accessibility of health care) were responsible for 44.4% of unmet needs in 2013 and 58.2% in 2014. The reasons related to the individual preferences of respondents contributed in 55.6% of cases in 2013 and in 41.7% of cases in 2014. The most dominant reason within the threatened accessibility was a financial reason, within the threatened availability were waiting lists for medical interventions and procedures, and within the threatened acceptability wish to wait and see if the state of health will improve and the problem will pass by itself..

Lithium Modulates the Chronic Stress-Induced Effect on Blood Glucose Level of Male Rats

In the present study we examined gross changes in the mass of whole adrenal glands and that of the adrenal cortex, as well as the serum corticosterone and glucose level of mature male Wistar rats subjected to three different treatments: animals subjected to chronic restraint-stress, animals injected with lithium (Li) and chronically stressed rats treated with Li. Under all three conditions we observed hypertrophy of whole adrenals, as well as the adrenal cortices. Chronic restraint stress, solely or in combination with Li treatment, significantly elevated the corticosterone level, but did not change the blood glucose level. Animals treated only with Li exhibited an elevated serum corticosterone level and blood glucose level. The aim of our study was to investigate the modulation of the chronic stress-induced effect on the blood glucose level by lithium, as a possible mechanism of avoiding the damage caused by chronic stress. Our results showed that lithium is an agent of choice which may help to reduce stress-elevated corticosterone and replenish exhausted glucose storages in an organism

Changes of c-myc expression in b16 melanoma cells induced by 8-chloroadenosine-3′, 5′-monophosphate and tiazofurin

The aim of this study was to investigate the in vitro effects of 8- chloroadenosine 3′, 5′-monophosphate (8-Cl-cAMP) and tiazofurin (TR) on the expression of c-myc gene in B16/F10 and B16/C3 mouse melanoma cells. Exponentially growing cells were treated with 8-Cl-cAMP or TR (5µmol - 25µmol) for 6h and 24h. The level of c-myc expression, estimated by RT-PCR, did not significantly change in B16/F10 cells after treatment with 8-Cl-cAMP or TR. Similar results were obtained in B16/C3 cells after treatment with 8-Cl-cAMP. The level of c-myc expression has shown a significant increase in B16/C3 cells after treatment with TR. Further studies of these agents will lead to better understanding of molecular mechanisms of their action.Physical chemistry 2004 : 7th international conference on fundamental and applied aspects of physical chemistry; Belgrade (Serbia); 21-23 September 200

Non-isothermal dehydration of equilibrium swollen radiolytically sinthesized Fe3O4 – PVA ferrogel nanocomposite

In this study, the Fe3O4 - PVA ferrogel nanocomposite was synthesized by gamma irradiation. Obtained ferrogel had greater swelling capacity and activation energy of dehydratation as measured by thermogravimetric analysis under non-isothermal conditionsPhysical chemistry 2008 : 9th international conference on fundamental and applied aspects of physical chemistry; Belgrade (Serbia); 24-28 September 200

Annealing effects on the properties of TiN thin films

The structure, absorption coefficient and electrical resistivity studies on TiN thin films are presented. The film of thickness 240 nm was grown on Si (100) substrate by DC reactive sputtering at an average deposition rate of similar to 8 nm/min. After deposition the samples were annealed for 1 h at 600 degrees C and 2 h at 700 degrees C in nitrogen ambient and vacuum furnace, respectively. Structural characterizations were performed by Rutherford backscattering spectrometry (RBS), X-ray diffraction (XRD) and transmission electron microscopy (TEM). The optical properties were investigated by spectroscopic ellipsometry while a four point probe was used for electrical characterization. It was found that the post-deposition annealing of the films did not cause any variation in stoichiometry, but strongly affects the structural parameters such as lattice constant, micro-strain and grain size. The observed increase in the grain size after annealing leads to significantly lower value of the coefficient of absorption. These changes could be directly correlated with variation of electrical properties of TiN thin films

Ion-beam irradiation effects on reactively sputtered CrN layers

This paper presents a study of microstructural changes induced in CrN layers by irradiation with 120 keV argon ions. The layers were deposited on (100) Si wafers, at different nitrogen partial pressures (2×10^-4, 3.5×10^-4 and 5×10^-4 mbar), to a total thickness of 260–280 nm. During deposition the substrates were held at 150°C. After deposition the samples were irradiated with argon ions to the fluences of 1×10^15 and 1×10^16 ions/cm2, under the vacuum of 7×10^-6 mbar. Characterisation of the samples structure and morphology were performed by X-ray diffraction (XRD) analysis and cross-sectional transmission electron microscopy (XTEM), and the concentration profiles were determined by Rutheford backscattering (RBS) spectrometry. It was found that the layer composition strongly depends on the nitrogen partial pressure during deposition. A pure stoichiometric CrN phase was achieved for the highest nitrogen partial pressure (5×10^-4 mbar). Argon ions irradiation induces microstructural changes in the CrN layers such as variation of the lattice constants, micro-strain and mean grain size.Paper presented at 4th Serbian Conference on Electron Microscopy, Belgrade, Serbia, 201

Argon Irradiation Effects on the Structural and Optical Properties of Reactively Sputtered CrN Films

The present study deals with CrN films irradiated at room temperature (RT) with 200 keV Ar+ ions. The CrN layers were deposited by d.c. reactive sputtering on Si (100) wafers, at nitrogen partial pressure of 5x10(-4) mbar, to a total thickness of 280 nm. The substrates were held at 150 degrees C during deposition. After deposition the CrN layers were irradiated with 200 keV Ar+ ions to the fluences of 5x10(15) - 2x10(16) ions/cm(2). Structural characterization was performed with Rutherford backscattering spectroscopy (RBS), cross-sectional transmission electron microscopy (XTEM) and X-ray diffraction (XRD). Spectroscopic ellipsometry measurements were carried out in order to study optical properties of the samples. The irradiations caused the microstructrual changes in CrN layers, but no amorphization even at the highest argon fluence of 2x10(16) ions/cm(2). Observed changes in microstructure were correlated with the variation in optical parameters. It was found that both refractive index and extinction coefficient are strongly dependent on the defect concentration in CrN layers