BRAF(V600E), the most commonly mutated oncogene in melanoma, is found in about half of patients. By hyperactivating the MAPK pathway, this mutation promotes cell growth and proliferation. Melanocytic BRAF(V600E) alone, however, is insufficient to cause melanoma and rather promotes the development of benign nevi (moles). The goal of our initial studies was to better understand how genetic and environmental risk factors interact with the BRAF(V600E) oncogene to induce melanoma. The two most prominent risk factors for melanoma development are exposure to ultraviolet (UV) radiation and pale skin pigmentation; particularly in the case of individuals with the “redhead” phenotype, who carry inactivating mutations in the MC1R G-protein coupled receptor. It has commonly been thought that redheads are at highest risk for melanoma development due to poor protection from genotoxic UV radiation from the sun. Using a melanocyte-specific, inducible Braf(V600E) mouse model, we have shown that an inactivating mutation in Mc1r which causes a redhead phenotype in mice, confers a significant UV-independent elevation in melanoma risk, relative to black and albino animals. The mechanism of accelerated UV-independent oncogenesis was found to be dependent on the synthesis of the red/yellow pheomelanin pigment. While these experiments were on-going, a novel small molecule inhibitor of the BRAF(V600E) oncogene, vemurafenib, began showing promising results in clinical trials. The observation that half of patients were experiencing significant tumor regression was unprecedented, but was soon followed by vemurafenib-resistant disease progression. Based on the fact that acquired drug resistance is a major obstacle to good therapeutic outcomes, we began investigating mechanisms of BRAF inhibitor resistance. A panel of BRAF(V600E) human melanoma cell lines that were initially sensitive to PLX4720 (a pre-clinical analog of vemurafenib), were chronically treated with the oncogenic BRAF inhibitor until resistance developed. These paired resistant and sensitive cell lines were characterized in terms of drug sensitivity and activation of cell signaling pathways. Multiple different patterns of drug resistance were found. The diversity of resistance mechanisms in these studies agrees with the diversity which others have found in the literature, suggesting that melanoma cells may be uniquely adaptable to circumventing BRAF(V600E) oncogene addiction.

The Proopiomelanocortin gene, (POMC), produces many biologically active peptides including the endogenous opioid, β-endorphin, and the melanocortins: α-Melanocyte Stimulating Hormone, (αMSH), γMSH, βMSH and Adrenocorticotropic Hormone, (ACTH). βendorphin is released by the brain in response to stress or injury and is a potent analgesic. Melanocortins are well known for regulating pigmentation, metabolism, and cortisol levels. Additionally, opioids and melanocortins are known to have opposing actions in several settings including the regulation of pain and metabolism. The Melanocyte Stimulating Hormones are expressed in the skin where they bind the Melanocortin 1 Receptor on melanocytes and promote pigmentation. It has been reported that β-endorphin is also produced in the skin, however it was not believed to have a central effect. In this thesis I show that expression of these peptides in the skin is reflected in blood levels and affects nociception and behavior.

The demand for higher bandwidth and higher speed driven by semiconductor technology development draws a great deal of research efforts devoted to the development of high speed data communication. Challenges on electrical copper interconnects at high frequency make optical interconnect technologies become a promising alternative to conventional electrical interconnects at different levels. This doctoral dissertation describes polymer based nano- and micro-photonic devices for three-dimensional optical interconnects. Two areas are focused, (1) polymer based two-dimensional (2D) and three-dimensional (3D) photonic crystal fabrication and simulation for laser beam steering applications, (2) polymer based optical waveguide array and shared bus waveguide with embedded 45° micro-mirrors for board level optical interconnects. A three-dimensional (3D) face-centered cubic (FCC) type polymer based photonic crystal using the polymer material SU-8 was simulated and successfully fabricated using a polygonal prism based holographic fabrication method. The theoretical study of polymer based photonic crystals was carried out for laser beam steering, which is based on the superprism effect. Horizontally stacked two-dimensional (2D) photonic crystal was fabricated by a double exposure holographic interference method. The k-vector superprism effect, the principle for beam steering, was studied in detail through EFC (Equi-frequency Contour) analysis. A polymer based optical waveguide array with embedded 45° micro-mirrors for board level optical interconnects was prepared using a Ni metal hard mold by a UV imprint technique. A nickel based metal mold with 45º tilted surfaces on both ends of the channel waveguide was prepared through the electroplating process. To obtain a precise 45º tilted angle, a 50µm thick SU-8 layer was exposed under de-ionized water. High speed optical testing (10Gb/s) was carried out on the polymeric optical waveguide array with embedded 45º micro-mirrors on flexible substrate for out-of-plane optical interconnects. A polymer based 3-to-3 shared optical bus waveguide with opposite 45º micro-mirrors was designed and fabricated using the metallic hard mold method. The Ni metal hard mold was successfully prepared using the Ni electroplating method. This metallic hard mold provides a convenient way to fabricate the polymeric optical bus waveguide devices through the imprint technique. / text

Gas phase chemiluminescence nitrogen specific detector (CLND)is used for the direct analysis of underivatized nitrogen-containing components such as alkylamines that can not be detected by the so called universal HPLC detector, the UV detector. However, alkali metal hydroxides can not be used as mobile phase constituents with the CLND because they form non volatile particulate combustion products that foul the detector. Therefore, trimethylsulfonium hydroxide (TMSOH) has been selected as a strong base for use with the CLND, because its combustion products, CO2, H2O and SxOy are volatile. An alkali-stable zirconia-based column was used and coupled to the CLND. Zirconia-based columns are mechanically and hydrolytically more stable than silica-based columns, which have a working pH range from 3 to 8 only. Zirconia-based columns can be used at a pH from 1 to 14 and can be used at temperatures up to 200˚C. The separation of amines was carried out at high pH values where the amino groups were deprotonated. Primary, secondary, tertiary and quaternary amines were separated using a pH=13.7 mobile phase that contained only TMSOH, methanol and water. Good peak shapes were observed for all, except n-alkylamines and samples that contained both amino groups and alcohol groups.

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Viena svarbiausių augalų genetikos problemų yra jų atsparumas abiotiniams veiksniams. Tokie veiksniai kaip šaltis, ozonas ar ultravioletinė spinduliuotė sukelia oksidacinį stresą ir ROS formavimąsi augalų ląstelėse. Visi augalai turi antioksidantines sistemas, vykdančias ROS detoksikaciją. Žalioji kreisvė (Crepis capillaris L.(Wallr.)) ir atsparūs šalnoms bulvių somatiniai asimetriniai hibridai buvo tiriami atsparumui abiotiniams veiksniams, įvertinant morfometrinius, biocheminius ir genetinius rodiklius po UV-B –(2, 4, 8 kJ/m2) ir ozono (40 ir 80 ppb) poveikio. Nustatyta, kad jautrūs stresiniams veiksniams yra morfometriniai parametrai- žalia ir sausa biomasė, lapų plotas ir skaičius. Biocheminiai parametrai, tirpių baltymų kiekis ir antioksidantinio fermento superoksido dismutazės (SOD) aktyvumas, tirtuose augaluose proporcingai didėjo priklausomai nuo dozės dydžio. Padidėjęs antioksidantinio SOD fermento aktyvumas po abiotinių veiksnių poveikio vertintinas kaip augalo adaptacinis atsakas į UV-B sukeltą oksidacinį stresą. Tiriant atsparumą šalčiui nustatyta, kad atsparus šalčiui bulvių hibridas H269, įgijo iš donoro Solanum commersonii DNR fragmentą, susijusį su padidinta mitochondrijų elektronų pernašos sistemos genų raiška, kuri siejama ROS koncentracijos reguliavimu ląstelėje ir šalčio atsako genų reguliavimu branduoliniame genome. Parodėme, kad atsparumai abiotiniams veiksniams- ozonui, UV-B ir šalčiui augaluose funkcionuoja kaip vientisa gynybos sistema, apimanti... [toliau žr. visą tekstą] / One of the major issues of plant genetics is their tolerance and resistance to abiotic factors. Low temperatures, ozone and ultraviolet radiation are the factors that increase ROS formation and oxidative stress in a plant cell. All plants have effective antioxidant systems to detoxify ROS. Researches were carried out to evaluate the resistance of Smooth Hawksbeard (Crepis capillaris L. (Wallr)) and frost resistant asymmetric somatic potato hybrids to abiotic stresses, estimating the morphometrical, biochemical and genetic data after treating the plants with UV-B (2, 4, 8 kJ/m2) and ozone (40 ir 80 ppb) doses. Test data shows, that morphometrical parameters such as fresh and dry weight along with leaf area and count are very sensitive to stress factors. Biochemical parameters such as soluble protein content and the activity of antioxidant enzyme superoxide dismutase (SOD) have risen pro rata from the dose given. The increment of SOD after treating the plants with simulated abiotic factors is annotated as adaptation to UV-B stress. Analysis of the cold acclimation has shown, that potato hybrid H269 has gained DNA fragment from the donor species Solanum commersonii. This fragment is associated with over-expressed genes of mitochondrial electron transfer system which regulates the concentration of ROS in the cell and controls the expression of nuclear genes related with cold-acclimation. With this analysis we have showed that plant resistance to abiotic factors such as ozone... [to full text]

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Suspended titanium dioxide (TiO2) nanoparticles produce hydroxyl radicals (•OH) that synergistically aid in disinfection when irradiated with UV light. To exploit the benefits of TiO2 photocatalysis without having to remove them, TiO2 was deposited onto 3M Company’s nanostructured thin film (NSTF). •OH production by suspended and TiO2-NSTF was determined by para-chlorobenzoic acid (pCBA), a •OH probe compound. Both techniques of introducing TiO2 to the samples produced equivalent •OH without forming bromate, a regulated byproduct in drinking water at all UV and TiO2 levels. Formation of bromate by ozone in brackish water and seawater were used to compare the disinfection byproduct (DBP) formation between the disinfection methods. Additionally, monitoring bromate, typically performed by ion chromatography, is complicated by chloride and other anions present in brackish water or seawater. Thus, a spectrophotometric method to measure bromate in saline systems is introduced.

No description available.

Endocrine disrupting compounds (EDCs) in the environment have recently emerged as a major issue in Canada and around the globe. The primary objective of this thesis was to investigate the fate of EDCs in two wastewater treatment processes, membrane bioreactors (MBRs) and ultraviolet (UV) disinfection. Two submerged MBR systems using hollow fiber membranes from two membrane manufacturers were tested. The results from a bench-scale and a pilot scale MBR for the treatment of swine wastewater with high concentration of EDCs showed that over 94% of the estrogenic activity (EA) in the influent was reduced through the MBR process. Biological degradation was the dominant removal mechanism for the removal of EDCs in MBRs. Over 85% of the influent EA was reduced by biodegradation through the MBR process. The other MBR system was built to study the removal mechanisms of two estrogens in a hybrid MBR with the addition of powdered activated carbon (PAC). The effects of PAC dosing on MBR overall performance was studied as well. It was found that PAC dosing could increase the removal rates of 17β-estradiol (E2) and 17α-ethinylestradiol (EE2) by 3.4% and 15.8%, respectively and result in a slower rate of trans-membrane pressure (TMP) increase during MBR operation, which could significantly reduce the operating cost for membrane cleaning and/or replacement. The operating cost for PAC dosing could be offset by the benefit achieved from reducing the cost for membrane maintenance. The slower rate of TMP increase in the PAC-MBR was associated with the lower concentrations of soluble extracellular polymeric substances and colloidal organic compounds in the PAC-MBR sludge. The degradation kinetics of three estrogens, estrone (E1), E2, and EE2 in de-ionized water by UV irradiation was studied. The experimental results showed both the apparent concentrations and overall EA of all three investigated estrogens in water decreased with direct UV irradiation. To further study the impact of UV on the overall EA of wastewater, the EA of pre-UV and post-UV samples from five wastewater treatment plants were measured in both liquid and solid phase by Yeast Estrogen Screen assay. It was found that the EA of wastewater decreased after UV disinfection in three of the investigated plants whereas it increased in the other two plants. This observation needs to be further studied because it might have significant impacts on the application of UV systems for wastewater disinfection.