<p>Radiotherapy is one of the most common therapeutic methods for treatment of many types of cancer. Despite many decades of development and experience there is much to improve, both in efficacy of treatment and to decrease the incidences of adverse healthy tissue reactions. Around 20 % of the radiotherapy patients show a broad range in the severity of normal tissue reactions to radiotherapy, and dose limits are governed by severe reactions in the most radiosensitive patients (< 5 %). Identification of patients with low, moderate or high clinical radiosensitivity before commencing of radiotherapy would allow individual adaptation of the maximum dose with an overall increase in the cure rate. Characterization of factors that may modify the biological effects of ionizing radiation has been a subject of intense research efforts. Still, there is no assay currently available that can reliably predict the clinical radiosensitivity. The aim of this work has been to investigate the role of oxidative stress in individual radiosensitivity and evaluate novel markers of radiation response, which could be adapted for clinical use.</p><p>8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG), a general marker of oxidative stress, is one of the major products of interaction of ionizing radiation with DNA and the nucleotide pool of the cell. As 8-oxo-dG is highly mutagenic due to incorrect base pairing with deoxyadenosine, various repair mechanisms recognize and remove 8-oxo-dG. The repaired lesions are released from cells to the extracellular milieu (serum, urine and cell culture medium) where they can be detected as markers for free radical reactions with the nucleic acids.</p><p>Significant variations in background levels as well as in radiation induced levels of 8-oxo-dG in urine have been demonstrated in breast cancer patients (paper 1). Two major patterns were observed: high background and no therapy-related increase vs. low background and significant increase during radiotherapy for the radiosensitive and non radiosensitive patients respectively.</p><p>Studies in paper 2 indicated major contribution of the nucleotide pool to the extracellular 8-oxo-dG levels. The results also implicated induction of prolonged endogenous oxidative stress in the irradiated cells. RNA “knock-down” experiments on the nucleotide pool sanitization enzyme hMTH1 in paper 3 lend further experimental evidence to this assumption.</p><p>The applicability of 8-oxo-dG as a diagnostic marker of oxidative stress was demonstrated in paper 4. Studies on dialysis patients revealed a good correlation between inflammatory responses (known to be associated with persistent oxidative stress) and extracellular 8-oxo-dG.</p><p>In summary, our results confirm that extracellular 8-oxo-dG is a sensitive <i>in vivo</i> biomarker of oxidative stress, primarily formed by oxidative damage of dGTP in the nucleotide pool with a potential to become a clinical tool for prediction of individual responses to radiotherapy.</p>

Radiotherapy is one of the most common therapeutic methods for treatment of many types of cancer. Despite many decades of development and experience there is much to improve, both in efficacy of treatment and to decrease the incidences of adverse healthy tissue reactions. Around 20 % of the radiotherapy patients show a broad range in the severity of normal tissue reactions to radiotherapy, and dose limits are governed by severe reactions in the most radiosensitive patients (&lt; 5 %). Identification of patients with low, moderate or high clinical radiosensitivity before commencing of radiotherapy would allow individual adaptation of the maximum dose with an overall increase in the cure rate. Characterization of factors that may modify the biological effects of ionizing radiation has been a subject of intense research efforts. Still, there is no assay currently available that can reliably predict the clinical radiosensitivity. The aim of this work has been to investigate the role of oxidative stress in individual radiosensitivity and evaluate novel markers of radiation response, which could be adapted for clinical use. 8-Oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG), a general marker of oxidative stress, is one of the major products of interaction of ionizing radiation with DNA and the nucleotide pool of the cell. As 8-oxo-dG is highly mutagenic due to incorrect base pairing with deoxyadenosine, various repair mechanisms recognize and remove 8-oxo-dG. The repaired lesions are released from cells to the extracellular milieu (serum, urine and cell culture medium) where they can be detected as markers for free radical reactions with the nucleic acids. Significant variations in background levels as well as in radiation induced levels of 8-oxo-dG in urine have been demonstrated in breast cancer patients (paper 1). Two major patterns were observed: high background and no therapy-related increase vs. low background and significant increase during radiotherapy for the radiosensitive and non radiosensitive patients respectively. Studies in paper 2 indicated major contribution of the nucleotide pool to the extracellular 8-oxo-dG levels. The results also implicated induction of prolonged endogenous oxidative stress in the irradiated cells. RNA “knock-down” experiments on the nucleotide pool sanitization enzyme hMTH1 in paper 3 lend further experimental evidence to this assumption. The applicability of 8-oxo-dG as a diagnostic marker of oxidative stress was demonstrated in paper 4. Studies on dialysis patients revealed a good correlation between inflammatory responses (known to be associated with persistent oxidative stress) and extracellular 8-oxo-dG. In summary, our results confirm that extracellular 8-oxo-dG is a sensitive in vivo biomarker of oxidative stress, primarily formed by oxidative damage of dGTP in the nucleotide pool with a potential to become a clinical tool for prediction of individual responses to radiotherapy.

The semi-enclosed and brackish Baltic Sea has become heavily polluted by nutrients, anthropogenic organic and inorganic chemicals via human activities. Persistent organic pollutants (POPs) have been thoroughly investigated due to their linkage to toxic effects observed in Baltic biota. There has been far less focus on semi-persistent pollutants e.g. naturally produced oraganohalogen compounds (NOCs) and their disturbances in the environment. This thesis is aimed on assessment of levels and trends of naturally produced brominated compounds in Baltic biota; more specifically on hydroxylated polybrominated diphenyl ethers (OH-PBDEs), methoxylated PBDEs (MeO-PBDEs) and polybrominated dibenzo-p-dioxins (PBDDs). These, NOCs, may originate from production in algae and cyanobacteria. OH-PBDEs and MeO-PBDEs may also be formed as metabolites of polybrominated diphenyl ethers (PBDEs), i.e. well-known commercial flame retardants. High levels of OH-PBDEs, MeO-PBDEs and PBDDs are shown within Baltic biota (cyanobacteria, algae, mussels, fish), often in much higher concentrations than PBDEs which are possible anthropogenic precursors of OH- and MeO-PBDEs. The levels of OH-PBDEs, MeO-PBDEs and PBDDs are higher in the Baltic Sea than on the west coast of Sweden. Temporal and seasonal variations show fluctuations in concentrations of OH-PBDEs, MeO-PBDEs and PBDDs, possibly related with macroalgal life-cycles. OH-PBDEs, MeO-PBDEs and PBDDs are present in several filamentous macroalgae species, but considering the levels quantified, the time of peak exposure and the species life-cycle the macroalgae, Pilayella, Ceramium and Cladophora are suggested as major natural producers of OH-PBDEs and PBDDs. The high levels of OH-PBDEs, MeO-PBDEs and PBDDs in the Baltic Sea may affect numerous organisms in the ecosystem. The toxic effects of OH-PBDEs and PBDDs are of particular concern. This thesis stress the importance of assessing and monitoring these substances, since the exposure to OH-PBDEs and PBDDs, during summer, may cause acute effects in Baltic fish and wildlife. / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: In press. Paper 4: Manuscript.

Oxidation of organic species, in particular dicarboxylic and humic acids, was investigated in ice. Products were analyzed by Proton Transfer Reaction Mass Spectrometry, ion and gas chromatography, and a Total Organic Carbon analyzer. Photolysis of succinic acid with H2O2, an OH precursor, produced malonic acid and malic acid, illustrating that diacids are subject to photochemical degradation in ice. First-order decay rate constants were an order of magnitude higher at room temperature (~23 °C) than in ice (-20 °C). A smaller difference was observed for malonic acid, a more soluble diacid, suggesting that partial segregation of H2O2 and succinic acid during freezing played an important role in the kinetics. VOCs, likely to be aldehydes and ketones, were produced from ice containing humic acid through heterogeneous ozonolysis and photooxidation which was enhanced by NO3-, an OH precursor. VOCs also formed from ice made from deionized water, likely through oxidation of organic contaminants.

The vitamin D metabolite 1,25(OH)2D3 has long been known to inhibit growth of prostate cancer cells and this mainly through a VDR-mediated pathway controlling target gene expression, resulting in cell cycle arrest, apoptosis and differentiation. Another major way in which 1,25(OH)2D3 inhibits cell growth in prostate cancer is via membrane-initiated steroid signalling, which triggers activation of signal cascades upon steroid binding to a receptor complex, leading to induction of genes regulating cell growth, proliferation and apoptosis. The main prostate cancer inhibiting membrane-initiated route is the TGFβ signalling pathway, elicited by the protein TGFβ. Two other important proteins downstream in this cascade are Smad2 and Smad3. In this study the early effects of 1,25(OH)2D3 on activated Smad2/3 levelsin PC-3 prostate cancer cells were examined. PC-3 cells were incubated for 3, 5, 10, 30 and 60 minutes as well as 38 hours both together with 1,25(OH)2D3 of the concentrations 10-10 and 10-7 M and without. Western Blots were then performed on supernatants from the cells treated followed by treatment of the membranes with primary antibodies against phosphorylated Smad2/3 C-terminal linker regions, alkaline phosphatase conjugated secondary antibodies and finally visualization with BCIP/ NBT tablets. As the downstream cascade protein JNK is a proposed activator of Smad2/3, this procedure was also repeated with a JNK inhibitor. This is a follow-up to an earlier study which examined the influence of 1,25(OH)2D3 on TGFβ levels using the same doses and time points and which found that 1,25(OH)2D3 initially lowered the level of active TGFβ, then increased it. The results of this study indicated a 1,25(OH)2D3 mediated induction of the same pattern in the levels of active Smad2 and 3, both with and without JNK inhibitor. The results did not indicate that 1,25(OH)2D3 activates the Smad2/3 C-terminal linker region via the JNK pathway.

The vitamin D metabolite 1,25(OH)2D3 has long been known to inhibit growth of prostate cancer cells and this mainly through a VDR-mediated pathway controlling target gene expression, resulting in cell cycle arrest, apoptosis and differentiation. Another major way inwhich 1,25(OH)2D3 inhibits cell growth in prostate cancer is via membrane-initiated steroid signalling, which triggers activation of signal cascades upon steroid binding to a receptor complex, leading to induction of genes regulating cell growth, proliferation and apoptosis. The main prostate cancer inhibiting membrane-initiated route is the TGFβ signalling pathway, elicited by the protein TGFβ. Another important protein downstream in this cascade is Smad2. In this study the early effects of 1,25(OH)2D3 on activated Smad2 levels in PC-3 prostate cancer cells were examined. PC-3 cells were incubated for 5, 10, 30 and 60 minutes as well as 24 and 40 hours both together with 1,25(OH)2D3 of the concentrations 10-10 and 107 M and without. An ELISA assay scanning for activated Smad2 was then performed on supernatants from both treated and untreated cells. This is a follow-up to an earlier study which examined the influence of 1,25(OH)2D3 on TGFβ levels using the same doses and similar time points and which found that 1,25(OH)2D3 initially lowered the level of active TGFβ, then increased it. The results of this study showed a statistically insignificant, time delayed 1,25(OH)2D3 mediated induction of the same pattern in the levels of active Smad2. / Project Work in Biomedicine, Advanced Level, 7.5 ECTS

Recent field campaigns have measured enhanced levels of NOx (NO+NO2) and HOx precursors (i.e., H2O2, CH2O, and HONO) that can not be accounted for by gas phase chemistry alone. Snowpack emission is now considered a source of these species. Therefore, the photochemistry in the polar boundary layer is now believed to be much more complex than initially thought. Field campaigns to Summit, Greenland in the summer of 2003 and the spring of 2004 have obtained the first measurements of peroxy (HO2+RO2) and hydroxyl (OH) radicals in the Artic boundary layer. Measurements were collected with a chemical ionization mass spectrometer (CIMS). A highly constrained (ie., O3, H2O, CH4, CO, j-values, NO, H2O2,CH2O, and HONO) 0-D steady-state model was employed in order to test our current understanding of photochemistry. HO2+RO2 measurements were in excellent agreement with model predictions for both spring and summer. OH measurements were in good agreement with spring model predictions but were a factor of two greater than summer model predictions. The role of snowpack emission is also addressed in a HOx budget performed on the spring campaign. Measurements of nitric acid (HNO3) and pernitric acid (HO2NO2) were obtained with the CIMS during the Antarctic Tropospheric Chemistry Investigation (ANTCI). The linkage between HOx and NOx chemistry is examined through partitioning of reactive nitrogen between HNO3 and HO2NO2. The possible impact of reactive nitrogen partitioning on nitrate ions (NO3-) at coring sites is also investigated.

The specific surface area reduction and pore size distribution coupled with N2 adsorption-desorption hysteresis isotherm were studied in the temperature range of 1400-1550¢J for periclase MgO powder having 0.1£gm in size and with face-centered cubic structure. The apparent activation energy of such a rapid coarsening-coalescence process for MgO powder was estimated as 181¡Ó3kJ/mol. The minimum temperature for sintering/coarsening/coalescence of submicron MgO particles was estimated to be near 1300¢J based on the extrapolation of steady specific surface area reduction rates to zero. Pulsed laser ablation (PLA) of periclase MgO powders in water was conducted under Q-switch mode and specified water height and water depth (10 mm) for an accumulation time of 5 and 20 minutes at 10 Hz. Such a PLA process has successfully synthesized nanosized and protonated MgO particles from Mg(OH)2 and lamellar precusors, implying the three phases may co-exist at high pressure and temperature conditions upon dynamic shock loading. A significant internal compressive stress up to 10 GPa was built up for the MgO but not the readily relaxed Mg(OH)2 nanocondensates. The lamellae-derived Mg(OH)2 tended to undergo a dehydroxylation process to become MgO following a specific crystallographic relationship, i.e. lamellar basal layer parallel to Mg(OH)2(0001) and MgO(111). The minimum band gap of the colloidal solution of MgO/Mg(OH)2/lamellae was lowered to ca. 5.2eV after the PLA process.

The resistance random access memory (RRAM) is one of the most popular of the next generation memories with the high operating speed, reliability and the smallest miniature size. RRAM has metal-insulator-metal structure that can greatly reduce the difficulty of entry, but the biggest problem is how to choose the insulator. We selected silicon-based materials to match the intergrated circuits manufacturing process. In this work, sputtering titanium doping in the silicon oxide thin film has a stable characteristic of resistance switching. By material analyzing, we found that supercritical carbon dioxide fluid (SCCO2) treatment can passivate the silicon oxide defect and the self-reduction of titanium oxide, but it also brought OH group into our thin film. So we observed the interface type characteristic of resistance switching. Using constant voltage sampling experiment extract the reaction rate constant (k) and the active energy, prove that the reaction is caused by OH injection. Double-layer structure with titanium-doped and carbon-doped silicon oxide RRAM promote lower operating current by hopping conduction, which is caused by graphite oxide doping. The Space-Charge Limited Current mechanism for high limited current is proven by COMSOL electric field simulation.

Exposure to atmospheric conditions results of deterioration in historic monuments. and their stones. Limestone conservation presents many problems that have to be investigated in detail. In this study, limestone deterioration and development of its conservation treatments were investigated through examination of the statues carved from karstic limestones in Nemrut Dag Monument. The decay mechanisms that had major roles in their deterioration during two thousand years of exposure to atmospheric conditions and the development of their conservation treatments involved several types of analyses that were carried out in the field and in the laboratory. Exposed surfaces of limestones having karstic veins, interior crack surfaces were examined and compared with relatively undeteriorated interior parts. Similar limestones from the geological formations nearby were artificially deteriorated by salt crystallization and were also examined for comparison. Standard physical and physicomechanical tests, petrographical analysis, XRD, SEM-EDX and FTIR were used during those examinations. Swelling nature of clays in limestones and their control were quantified by CEC measurements. The micro structure of limestone was observed to be composed of micritic calcite with karstic veins of sparitic calcite crystals. Some karstic zones were found to be preferred sites of dissolution and precipitation of calcium carbonate where swelling action of clays and widening of cracks occurred. Iron oxides that moved through those zones, as well as biological activity were also found to contribute to those phenomena. Preparation of high concentrations of nanodispersive calcium hydroxide solutions was achieved for the conservation treatments of the deteriorated limestone. Success of treatments with nanodispersive Ca(OH)2 solutions targeted to the decay zones were discussed in terms of their ability to control the swelling action of clays, carbonation of nanodispersive solution, and improvement in the physicomechanical properties of treated limestone.