O. Nešković

Using carbon nanotubes to induce micronuclei and double strand breaks of the DNA in human cells

Carbon nanotubes are unique one-dimensional macromolecules with promising applications in biology and medicine. Since their toxicity is still under debate, here we present a study investigating the genotoxic properties of purified single wall carbon nanotubes (SWCNTs), multiwall carbon nanotubes (MWCNTs), and amide functionalized purified SWCNTs on cultured human lymphocytes employing cytokinesis block micronucleus assay and enumeration of gamma H2AX foci as a measure of double strand breaks (DSBs) of the DNA in normal human fibroblasts. SWCNTs induce micronuclei (MN) formation in lymphocytes and decrease the proliferation potential (CBPI) of cells. In a fibroblast cell line the same dose of SWCNTs induces gamma H2AX foci 2.7-fold higher than in a control. Amide functionalized purified SWCNTs behave differently: they do not disturb the cell proliferation potential of harvested lymphocytes, but induce micronuclei to a higher extent than SWCNTs. When applied on fibroblasts, amide functionalized SWCNTs also induce gamma H2AX foci, 3.18-fold higher than the control. The cellular effects of MWCNTs display the broad spectrum of clastogenic properties seen as the highest incidence of induced lymphocyte micronuclei and anaphase bridges among nuclei in binucleated cells. Surprisingly, the incidence of induced gamma H2AX foci was not as high as was expected by the micronucleus test, which indicates that MWCNTs act as clastogen and aneugen agents simultaneously. Biological endpoints investigated in this study indicate a close relationship between the electrochemical properties of carbon nanotubes and observed genotoxicity.

Nanostructure and mineral composition of trabecular bone in the lateral femoral neck: implications for bone fragility in elderly women.

Despite interest in investigating age-related hip fractures, the determinants of decreased bone strength in advanced age are not clear enough. Hitherto it has been obscure how the aging process affects the femoral neck nanostructure and composition, particularly in the lateral subregion of the femoral neck, which is considered as a fracture-initiating site. The femoral bone samples used in this study were obtained at autopsy in 10 women without skeletal disease (five younger: aged 20-40 years, and five elderly: aged 73-94 years). Atomic force microscopy (AFM) was applied to explore the mineral grain size in situ in young vs. old trabecular bone samples from the lateral femoral neck. The chemical compositions of the samples were determined using inductively coupled plasma optical emission spectroscopy and direct current argon arc plasma optical emission spectrometry. Our AFM study revealed differences in trabecular bone nanostructure between young and elderly women. The mineral grain size in the trabeculae of the old women was larger than that in the young (median: 95 vs. 59nm), with a particular bimodal distribution: 45% were small grains (similar to the young) and the rest were larger. Since chemical analyses showed that levels of calcium and phosphorus were unchanged with age, our study suggests that during aging the existing bone mineral is reorganized and forms larger aggregates. Given the mechanical disadvantage of large-grained structures (decreased material strength), the observed nanostructural differences contribute to our understanding of the increased fragility of the lateral femoral neck in aged females. Moreover, increasing data on mineral grains in natural bone is essential for advancing calcium-phosphate ceramics for bone tissue replacement.

Atomic force microscopy characterization of the external cortical bone surface in young and elderly women: potential nanostructural traces of periosteal bone apposition during aging.

On the basis of the suggestion that bone nanostructure bears “tissue age” information and may reflect surface deposition/modification processes, we performed nanoscale characterization of the external cortical bone surface at the femoral neck in women using atomic force microscopy (AFM). The specific aims were to assess age-related differences in bone nanostructure and explore the existence of nanostructural traces of potential bone apposition at this surface. Our findings revealed that the external cortical surface represents a continuous phase composed of densely packed mineral grains. Although the grains varied in size and shape, there was a domination of small grains indicative of freshly deposited bone (mean grain size: young, 35 nm; old, 37 nm; p > 0.05). Advanced quantitative analysis of surface morphological patterns revealed comparable roughness and complexity of the surface, suggesting a similar rate of mineral particle deposition at the surface in both groups. Calcium/phosphorus ratio, a measure of bone tissue age, was within the same range in both groups. In summary, our AFM analyses showed consistent nanostructural and compositional bone features, suggesting existence of new bone at the periosteal bone surface in both young and elderly women. Considering observed age-related increase in the neck diameter, AFM findings may support the theory of continuous bone apposition at the periosteal surface.

TEMPERATURE-PROGRAMMED DESORPTION MASS SPECTROMETRIC STUDY OF THE SURFACE PROPERTIES OF GLASSY CARBON

The temperature-programmed desorption (TPD) method combined with mass spectrometric analysis has been applied to the characterization of glassy carbon (GC) and modified glassy carbon surfaces. Five different samples of GC were obtained from a commercial resol type of phenol formaldehyde resin that had undergone several treatments: the samples were either pure GC, GC treated with boron and phosphorus, or GC modified by metal (silver and palladium). Mass spectrometric analysis has shown that the thermal decomposition of surface oxide species results in desorption of H2O, CO and CO2, these being the major gas products. On the basis of the TPD spectra obtained, desorption energies have been calculated using a peak position method. The influence of CO2 oxidation on the surface properties of the samples was also investigated. It was concluded that metal deposition on a glassy carbon surface contributes to increased stability of surface oxide species, the most stable surface being one that has been modified by palladium deposition.

MASS SPECTROMETRIC STUDY OF THE IONIZATION AND DISSOCIATION OF SULPHUR HEXAFLUORIDE BY MONOENERGETIC ELECTRON IMPACT

The ionization and fragmentation of SF6 under electron impact has been examined by means of a trochoidal electron velocity selector and a mass spectrometer. The ionization efficiency curves of the positive ions were recorded and the appearance energies of different electronic states of these ions were determined. The experimental results were combined with auxiliary thermochemical data to evaluate the dissociation energies D°°(F5S–F) = 4.01 eV, D°°(F4S–2F) = 6.39 eV, D°°(F3S–3F) = 9.98 eV, the ionization energy of SF5, = 11.27 eV and heats of formation ΔH°f(SF5+) = 1.81 eV, ΔH°f(SF4+) = 4.12 eV, ΔH°f(SF3+) = 4.11 eV and ΔH°f(SF3) = − 5.13 eV.

Genotoxic Assessment of Carbon Nanotubes

Carbon nanotubes are unique one-dimensional macromolecules with promising application in biology and medicine. Since their toxicity is still under debate, here we describe an investigation of genotoxic properties of purified single-walled carbon nanotubes (SWCNT), multiwall carbon nanotubes (MWCNT), and amide-functionalized purified SWCNT. We used two different cell systems: cultured human lymphocytes where we employed cytokinesis-block micronucleus test and human fibroblasts where we investigate the induction of DNA double-strand breaks (DSBs) employing H2AX phosphorylation assay.

Forensic or archaeological issue: is chemical analysis of dental restorations helpful in assessing time since death and identification of skeletonized human remains?

In 2011, small mass grave with completely skeletonized remains was discovered in Belgrade suburb. An eyewitness claimed that skeletons belonged to German soldiers killed in WWII. Anthropologists were engaged to investigate whether the skeletal remains correspond to the indicated German group or represent more recent case requiring court trial. Numerous dental restorations were noticed. Owing to the fact that different dental materials were used in dental practice at certain times, the aim of this study was to explore whether analysis of dental restorations could help in identification and estimation of time since death. Inductively coupled plasma optical emission spectrometry revealed that dental fillings corresponded to copper amalgam, conventional silver amalgam, silicophosphate cement, and zinc phosphate cement. Chemical results combined with anthropological and historical facts suggest that the individuals lived before the 1960s in country with well‐developed dental service at that time. Therefore, chemical analysis of dental fillings was useful to distinguish between skeletal remains that are too old to be of forensic interest and the remains relevant to legal investigations.

Detection of elements and radioactivity in pellets from long-eared owls (Asio otus) inhabiting the city of Belgrade (Serbia)

In this study, we analysed pellets from long-eared owls (Asio otus) collected from four localities in Belgrade (Serbia). The pellets contained the remains of prey, namely voles (Arvicola terrestris) and field mice (Apodemus agrarius). The concentrations of 14 elements (Ca, P, Mg, Na, K, Fe, Zn, Sr, Ba, Mn, Ti, Cu, Si, B) were evaluated in whole pellets and in samples containing only bone tissue, which were dissected from the whole pellet. The increased levels of certain elements, including Mn, Zn, Ba, Cu and radioactive 40K, indicate contamination of the soil by various sources, such as industrial plants and agricultural practices. From the results presented in this article, we suggest that the analysis of owl pellets may indicate the quality of the local environment.

Formation of oxygen complexes in controlled atmosphere at surface of doped glassy carbon

The effects of boron and phosphorus incorporation in phenolic resin precursor to the oxidation resistance of glassy carbon have been studied. In order to reveal the nature and composition of the oxygen complexes formed at the surface of doped glassy carbon, under controlled atmosphere, the surface of the samples was cleaned under vacuum up to 1273 K. Specific functional groups, subsequently formed under dry CO2 or O2 atmosphere on the surface of boron-doped and phosphorus-doped glassy carbon samples, were examined using the temperature-programmed desorption method combined with mass spectrometric analysis. Characterization of surface properties of undoped and doped samples has shown that in the presence of either boron or phosphorus heteroatoms, a lower amount of oxygen complexes formed after CO2 exposure, while, typically, higher amount of oxygen complexes formed after O2 exposure. It has been concluded that the surface of undoped glassy carbon has a greater affinity towards CO2, while in the presence of either boron or phosphorus heteroatoms, the glassy carbon surface affinity becomes greater towards O2, under experimental conditions.

Energetics of the ionization and fragmentation of phosphorus oxyfluoride studied by electron-impact spectroscopy

The relative intensities and appearance energies of several positive ions in the mass spectrum of POF3 are reported together with an ionization-efficiency curve for F–, the only negative ion of significant intensity. Probable ion-fragmentation pathways are proposed together with the heats of formation of some positive ions and ionization energies for POF2 and POF. The appearance energies of various electronic states of the observed ions have been determined by deconvolution of the experimental ionization efficiency curves by the energy-distribution difference method.