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Komparativno in vitro ispitivanje efekata ugljeničnih nanocevi u normalnim i malignim ćelijama pluća / A comparative in vitro study of the carbon nanotubes on normal and cancer lung cellsJojić Nikola 05 February 2018 (has links)
<p>Ugljenične nanocevi (UNC) imaju sve veću primenu u elektronici, kompjuterskoj i optičkoj industriji, kao i u biomedicini. Dok proizvodnja jednoslojnih UNC nanocevi beleži sve veći rast poslednjih godina, rizik koji nosi izlaganje ovom nanomaterijalu ostaje nerazjašnjen. Oskudni i često kontradiktorni podaci o toksičnim efektima jednoslojnih UNC ukazuju na potrebu za daljim ispitivanjima. U našim istraživanjima ispitivane su promene u ćelijskom odgovoru kao i morfološke promene nakon delovanja jednoslojnih ugljeničnih nanocevi na ćelijskoj liniji humanih fetalnih fibroblasta pluća MRC-5 i ćelijskoj liniji humanog adenokarcinoma pluća A549. U ovoj studiji korišćene su jednoslojne ugljenične nanocevi koje su sadržale rezidualne nečistoće poput gvožđa. Citotoksičnost jednoslojnih UNC (engl. single-walled carbon nanotubes – SWCNT) je ispitivana kolorimetrijskim MTT testom. Tokom 24 i 48h niske koncentracije jednoslojnih ugljeničnih nanocevi (<250 μg/mL) pokazale su nisku toksičnost na proliferaciju i vijabilnost u obe ispitivane ćelijske linije. Ipak, pri visokim koncentracijama UNC (250-750 μg/mL) antiproliferativni efekat je bio blizu IC50 vrednostima. Na osnovu rezultata dobijenih MTT testom može se zaključiti da su maligne A549 ćelije osetljivije na delovanje jednoslojnih UNC u odnosu na normalne MRC-5 ćelije. Kombinacija ugljeničnih nanocevi sa prirodnim polifenolima (resveratrolom i proantocijanidolima) nije značajno uticala na citotoksičnost u MRC-5 ćelijama, za razliku od A549 ćelija gde je tretman kombinacijama umanjio toksičnost ugljeničnih nanocevi. Transmisionom elektronskom mikroskopijom ispitivan je efekat jednoslojnih ugljeničnih nanocevi na ćelijsku morfologiju i preživljavanje. Intracelularni agregati ugljeničnih nanocevi primećeni su u obe ćelijske linije, čime je potvrđeno da ugljenične nanocevi ulaze u ćelije. Imajući u vidu da nanomaterijali poput ugljeničnih nanocevi indukuju oksidativni stres i njime posredovanu apoptozu, na protočnom citometru je određivano prisustvo ćelija u apoptozi i nekrozi. Tretman ćelija sa jednoslojnim ugljeničnim nanocevima nije doveo do značajnog porasta broja apoptotskih ili nekrotičnih ćelija, što ide u prilog niskoj toksičnosti ovog nanomaterijala, odnosno ukazuje na alternativne mehanizme toksičnosti. Međutim kombinacija jednoslojnih ugljeničnih nanocevi sa antioksidantima, resveratrolom i proantocijanidolima indukuje veći procenat apoptoze i nekroze u odnosu na tretman samo sa nanocevima. Promene u ekspresiji gena praćene su lančanom reakcijom polimeraza (PCR). Komparativna analiza rezultata genske ekspresije MRC-5 i A549<br />ćelija nakon tretmana sa jednoslojnim ugljeničnim nanocevima pojedinačno i u kombinaciji sa antioksidantima ukazala je na kompleksnost i raznolikost biološkog odgovora ispitivanih ćelija. U našem istraživanju ispitivana je i promena aktivnosti enzima antioksidativne zaštite i količine glutationa u ćeliji. Primena jednoslojnih ugljeničnih nanocevi u MRC-5 ćelijama dovodi po smanjenja specifične aktivnosti enzima SOD i GR, povećava specifičnu aktivnost GPx i ne utiče na promenu specifične aktivnosti GST i količine glutationa u ćeliji Primena jednoslojnih ugljeničnih nanocevi u A549 ćelijama dovodi po smanjenja specifične aktivnosti enzima SOD, ne utiče na promenu specifične aktivnost enzima GR, GST i GPx, i dovodi do povećanja količine glutationa u ćeliji. Ćelijska vijabilnost, morfološke promene, redoks homeostaza i ekspresija ispitivanih gena bile su promenjene nakon tretmana sa jednoslojnim ugljeničnim nanocevima. Iako su dobijeni rezultati značajni za procenu toksičnosti ugljeničnih nanocevi, neophodna su dalja istraživanja koja treba da doprinesu boljem razumevanju toksičnih efekata ugljeničnih nanocevi.</p> / <p>Carbon nanotubes are being actively introduced in electronics, computer science, and optics as well as for various biomedical applications. While production of single-walled carbon nanotubes (SWCNT) has escalated in recent years, the knowledge on risk associated with exposure remains unclear. Contradictory data on the toxic effects of single-walled carbon nanotubes highlights the urgent need for further studies. In this study we investigated the alterations in cellular response along with morphological changes induced by single-walled carbon nanotubes in human lung fibroblast cell line MRC-5 and adenocarcinoma human alveolar basal epithelial cells A549. In this study we used SWCNT containing large amounts of residual metallic impurities such is iron, and the iron concentration increased in dose dependent manner in cells exposed to SWCNT. Cytotoxicity was evaluated by MTT assay and SWCNT showed little cytotoxic effect on the proliferation and viability of two cell lines tested at the concentrations used (<250 μg/mL) within 24 and 48h. However exposing both cell lines to high concentrations (250-750 μg/mL) resulted in near IC50 values. Based on MTT test SWCNT were more cytotoxic to A549 cell line. Cytotoxicity of SWCNT in combination with natural polyphenols (resveratrol and proanthocyanidins) did not noticeably affect the cytotoxicity of SWCNT to MRC-5 cells. However introduction of polyphenols did reduce the cytotoxicity of SWCNT to A549 cells. Transmission electron microscopy was used to complement cytotoxicity assays and to investigate the pathological effect of internalized SWCNT on cell morphology and survival. Intracellular bundles of CNTs, possibly aggregated/agglomerated were observed in both cell lines, confirming internalization after 24h exposure. Since nanoparticles like carbon nanotubes are toxic mainly because they cause oxidative stress, often associated with an increased apoptosis we checked for apoptotic and necrotic cells using flow cytometry. Incubation with SWCNT did not result in pronounced apoptosis or necrosis supporting its low toxicity and possibly alternative mechanism of cell damage. However incubation with SWCNT in combination with resveratrol and proanthocyanidins induced higher levels of both apoptosis and necrosis than SWCNT alone. Changes in gene expression following exposure to SWCNT were evaluated by polymerase chain reaction PCR array which indicated complex and diverse change in expression of genes involved in apoptosis, cell proliferation and oxidative stress. Finally we investigated the modulation of the antioxidant enzyme system and the changes in the cytosolic levels of GSH. SWCNT reduced the specific activity of SOD and GR enzymes, increased GPx activity. No changes in intracellular levels of GSH were observed in MRC-5 cell line. Same treatment in A549 cell reduced the specific activity of SOD, had no effect on GR, GST and GPx activity, but increased intracellular levels of GSH. Cell viability, morphologic changes, redox homeostasis and gene expression were affected by the presence of SWCNT. Although our findings are useful in predicting human response against SWCNT exposure, further study is needed for better understanding of the effects of SWCNT.</p>
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Dobijanje i karakterizacija 2D i 3D funkcionalnih materijala iz klase halkogenida dopiranih srebrom / Preparation and characterisation of 2D and 3D functional materials from the group of chalcogenides doped with silverČajko Kristina 13 July 2018 (has links)
<p>Predmet istraživanja ove doktorske disertacije su bila halkogenidna stakla iz sistema Ag<sub>x</sub>(As<sub>40</sub>S<sub>30</sub>Se<sub>30</sub>)<sub>100–x</sub> (x = 0, 0.5, 1, 2, 3, 4, 5, 10, 12, 13, 15 at. % Ag) – 3D forma i tanki filmovi preparirani iz prethodno sintetisanih stakala (x ≤ 5 at. % Ag) – 2D forma.<br />Utvrđena je oblasti amorfnosti u faznom dijagramu po odabranom preseku. Ispitivan je uticaj procentualnog udela srebra na fizičke karakteristike sintetisanih stakala i prepariranih tankih filmova koje su od značaja za primenu ovakvih materijala.<br />Izvršena je karakterizacija električnih, optičkih, strukturnih i termičkih osobina na osnovu kojih su izvedeni zaključci o uticaju i modifikaciji strukture As<sub>40</sub>S<sub>30</sub>Se<sub>30 </sub>halkogenidne matrice usled inkorporacije atoma srebra. Strukturna analiza ispitivanih sastava je potvrdila homogenost uzoraka sa manjom koncentracijom srebra (x ≤ 5 at. % Ag), dok je kod uzoraka sa većim procentualnim udelom ovog metala (x = 10, 13, 15 at. % Ag) pokazano da postoji fazna separacija. Kod sastava sa x = 13 i 15 at. % Ag potvrđena je prisutnost kristalnih centara AgAsSe<sub>2.</sub> Na osnovu rezultata dobijenih DSC tehnikom, pokazano je da pri zagrevanju balk uzoraka dolazi do delimične kristalizacije koja se odvija zapreminski sa dvodimenzionalnim i trodimenzionalnim rastomkristalnih centara. Optička i spektralna ispitivanja su pokazala da uvođenje Ag u matricu stakla As<sub>40</sub>S<sub>30</sub>Se<sub>30</sub> dovodi do smanjenja širine optički zabranjene zone i kod 3D i 2D uzoraka, kao i da svi sastavi ispoljavaju normalan oblik disperzije indeksa prelamanja. Takođe, rezultati Raman spektroskopije su ukazali na činjenicu da inkorporacija srebra u strukturnu mrežu ispitivanih stakala utiče na formiranje Ag–(S,Se)–As veza, odnosno uzrokuje formiranje novih strukturnih jedinica koje bi mogle uticati na provodljivost ovih sastava. Merenja električnih karakteristika 3D uzoraka izvršena su u jednosmernom i naizmeničnom režimu i pokazano je da koncentracija srebra ima značajan uticaj na električne osobine. Utvrđeni su različiti mehanizmi odgovorni za transport nosilaca naelektrisanja zavisno od koncentracije dopanta. Kompleksni impedansni spektri svih sastava su ukazali na prisustvo temperaturske zavisnosti procesa relaksacije, na ne idealan Debye–vski tip relaksacije, kao i negativni temperaturski koeficijent otpornosti koji je karakterističan za poluprovodnike.</p> / <p>The subject of this dissertation are chalcogenide glasses from the system Agx(As<sub>40</sub>S<sub>30</sub>Se<sub>30</sub>)<sub>100–x</sub> (x = 0, 0.5, 1, 2, 3, 4, 5, 10, 12, 13, 15 at. % Ag) – 3D form and thin films prepared from previously synthesised glasses (x ≤ 5 at. % Ag) – 2D form. The amorphous area in the phase diagram was determined by the selected tie– line. The influence of the silver percentage on the physical characteristics of the synthesized glasses and prepared thin films was investigated due to the importance of such materials for the application. Characterization of electrical, optical, structural and thermal properties has been performed, based on which conclusions on theinfluence and structure modification of the As<sub>40</sub>S<sub>30</sub>Se<sub>30</sub> chalcogenide matrix due to the incorporation of silver atoms have been derived. Structural analysis of the investigated samples confirmed the homogeneity of samples with a lower silver concentration (x ≤ 5 at.% Ag), while in samples with a higher percentage content of this metal (x = 10, 13, 15 at.% Ag) it was shown that there was a phase separation. The presence of crystal centers AgAsSe<sub>2</sub> was confirmed in the samples with x = 13 and 15 at. % Ag. Based on the results obtained with the DSC technique, it has been shown that by heating the samples, partial crystallization takes place voluminously, with two–dimensional and three–dimensional growth of crystalline centers. Opticaland spectral investigations have shown that the introduction of Ag into the glass matrix As<sub>40</sub>S<sub>30</sub>Se<sub>30</sub> leads to a reduction in the optical band gap in both 3D and 2D samples, and that all compositions exhibit a normal dispersion of index of refraction. Also, the Raman spectroscopy results pointed to the fact that the incorporation of silver into the structural network of the investigated samples influences the formation of Ag–(S,Se)–As structures that is, causes the formation of new structural units that could affect the conductivity of these compositions. Measurements of the electrical characteristics of the 3D samples were performed in DC and AC regime and it was shown that silver concentration has a significant effect on electrical properties. Different mechanisms that are responsible for the transport of charge carriers depending on dopant concentration were determined. The complex impedance spectra of all compositions indicated the presence of the temperature dependence of the relaxation process, the non–Debye relaxation and the negative temperature coefficient of resistance which is characteristic of the semiconductors.</p>
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Kriminalistinio medžiagų ir gaminių tyrimo galimybės: problemos ir perspektyvos / Possibilities of criminalistic investigation of materials and items: problems and perspectivesGudauskaitė, Aurelija 23 December 2014 (has links)
Kriminalistinis medžiagų ir gaminių tyrimas suprantamas kaip procesiškai reglamentuotas ekspertų ar specialistų atliekamas kriminalistinis medžiagų ir gaminių tyrimas, siekiant ištirti nusikalstamos veikos pėdsakus ir daiktinius įrodymus, ir reikalaujantis mokslo, technikos, meno ar kitų specialių žinių, ir kurio rezultatai įforminami ekspertizės akte arba specialisto išvadoje. Kriminalistinis medžiagų ir gaminių tyrimas apima platų ratą atskirų ekspertizės (tyrimų) rūšių, išskirtų pagal objekto kriterijų. Tai pluoštų, dirvožemio, naftos, alkoholinių skysčių, dažų ir kitos ekspertizės (tyrimai). Medžiagų ir gaminių tyrimui gali būti keliami įvairūs uždaviniai, kurie yra grupuojami į identifikacinius ir neidentifikacinius (klasifikacinius ir diagnostinius). Žinant kokie yra kiekvienai tyrimo rūšiai keliami uždaviniai, galima numanyti ir kokios yra ekspertinės galimybės: kokius faktus gali patvirtinti ar paneigti atliktas tyrimas. Ekspertizės (tyrimo) skyrimo teisinis pagrindas yra būtinumas atlikti specialių žinių reikalaujantį specialų tyrimą nusikalstamos veikos aplinkybėms nustatyti. Medžiagos tyrimui kokybiškas rengimas yra svarbi ekspertinio tyrimo atlikimo prielaida. Tyrimo objektai gali būti surandami tiek atliekant jų paiešką, tiek juos surandant. Suradus tyrimo objektus, itin svarbus objektų paėmimo ir pakavimo taisyklių, numatytų LTEC leistose metodinėse rekomendacijose bei kituose leidiniuose, laikymasis. Nuo to priklauso, ar bus išnaudotos visos ekspertinės... [toliau žr. visą tekstą] / The criminalistic investigation of materials and items is an investigation regulated by process rules and performed by experts or specialists seeking to examine criminal acts and material evidence which requires science, technique, art or other special knowledge and which results are formalized in an expert act or specialist opinion. Criminalistic investigation of materials and items is combined out of a wide range of independent expertise (object investigation) types which are distinguished by the object criteria. It includes fibres, soil, oil, alcoholic liquids, dies and other expertise (object investigation). There are various tasks being formulated for the investigation of materials and items, which are grouped into identificational and non-identificational tasks (the previous group includes both diagnostically and classification tasks). The awareness of what are the main tasks for every independent investigation allows perceiving what the expertise possibilities are: what facts can be approved or denied by the investigation. The legal background of the imposition of expertise (object investigation) is the necessity to perform a special investigation which requires the special knowledge being used in order to determine the circumstances of the criminal act. The qualified arrangement of the investigative material is an essential premise of the execution of the investigation. The investigative objects can be found in both ways, by performing the search of them or by... [to full text]
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