A new format for microfluidic based DNA biosensors is presented in which the biorecognition element (single stranded DNA probes) is immobilized as a continuous density gradient of probes along the length of a microfluidic channel instead of a standard array format commonly used in microarray technologies or DNA based biosensors. The development of continuous density gradients of immobilized probe was achieved by electrokinetically subjecting probes that were terminated with an appropriate functional group for a surface coupling reaction to increasing convective velocity along the length of the microfluidic channel. This gradient format was able to discriminate between a fully complementary target and one containing 3 BPM based on the spatial pattern of hybridization for picomole quantities of DNA targets. Temperature mediated destabilization of DNA hybrids demonstrated that the density of immobilized probes plays an important role in the thermodynamic stability of DNA hybrids. In addition, it was found that efficiency, selectivity and melt temperature of DNA hybrids for surface based hybridization is dependent on the density of the probe molecules.

The main goal of this research is to provide a novel bioanalytical approach to better understand α-synuclein (AS) aggregation linked to Parkinson’s disease (PD) and characterize the implications of contributing factors such as the presence of metal ions and potential therapeutics that would inhibit or reverse AS fibrillation. Current bioanalytical techniques have reported the fibrillation process of AS however, the detection of prefibrillar formation or the nucleation phase of AS has yet to be characterized. This research aimed to address this issue and monitor the primary stages of AS fibrillation from natively soluble monomer to fibrillar aggregates. The electrochemical measurement of these processes utilized the intrinsic electroactivity of 4 tyrosine (Tyr) residues in AS observed at ~0.6 V (vs. Ag/AgCl) to monitor its early fibrillation kinetics. The research presented here provided valuable evidence of the conformational changes attributed to prefibrillar forms of AS.

Passive air samplers (PAS) using XAD-resin were deployed at a wide variety of sites around the world for four years to asses the spatial and temporal trends of legacy organochlorine pesticides (OCPs) and current use pesticides (CUPs) in the global atmosphere. Legacy OCPs are prevalent in developing countries, whereas certain CUPs dominate in North America and Europe. OCP levels are declining world wide. Concentrations from the XAD-based PAS agreed with those from polyurethane foam (PUF) disk PAS within a factor of 4 for most sites. The comparison revealed that the sampling rate of the PUF-based PAS is more dependent on wind speed, whereas that of the XAD-PAS has a higher dependence on temperature. Analysis of PAS deployed across arid, subtropical Botswana showed that recent use has more impact on present day air concentrations than historical use. Year-long measurements by high volume air sampling in Botswana yielded higher HCB levels in winter, and higher endosulfan levels in summer. Those variations are neither related to temperature fluctuation nor seasonal hydrological events, and are therefore more likely caused by pesticide usage pattern. Very low levels of OCPs were found in the warm, dry, low organic matter soils of Botswana, including in soils from historical use areas. Such soils appear to have a low long-term storage capacity for pesticides allowing for rapid volatilization. They are thus not long-term sources of pesticides to the atmosphere. Endosulfan sulfate levels were observed to increase in tropical soils with increasing elevation. Water samples from high altitude cloud forests in Costa Rica contained very low concentrations of CUPs and it is unlikely that that those levels pose a threat to amphibians in cloud forests. Laboratory experiments and literature analysis showed that the inert gas stripping method for the determination of air-water partitioning equilibria is susceptible to surface sorption artifacts for chemicals with a interface-air partition coefficient log (KIA/m) > -3. Using larger bubbles reduces the surface-to-volume ratio and produces accurate air-water partition coefficients for chemicals with log (KIA/m) < -1.2.

The thickness shear mode acoustic wave (TSM) sensor, operated in a flow-through format, has been widely used in bioanalytical research. My research is mainly focused on the study of surface-attached biomolecules and cells using the TSM sensor, including lesions in DNA, conformational change of calmodulin, as well as the properties and attachment of rat aortic smooth muscle cells. Aldehydic apurinic or apyrimidinic sites (AP sites) that lack a nucleobase moiety are one of the most common forms of toxic lesions in DNA. In this work, synthesized oligodeoxyribonucleotides containing one, two, or three abasic sites were hybridized to complementary sequences immobilized on the gold electrode of the TSM device by affinity binding. The influence of AP sites on local base stacking energy and geometry caused a dramatic destabilization of the DNA duplex structure, which was detected by the TSM sensor. The signals detected by TSM correlated well with the thermostability of DNA duplexes in solution. The results indicate that both the number of sites and their localization in the double-stranded structure influence the stability of a 19 b.p. duplex. TSM was also used to detect the binding of ions or peptides to surface-attached calmodulin. The interaction between calmodulin and ions induced an increase in resonant frequency and a decrease in motional resistance. In addition, these signal changes were reversible upon washing with buffer. The response was interpreted as a decrease in surface coupling induced by exposure of hydrophobic domains on the protein, and an increase in the length of calmodulin by approximately 3 Å. In addition, the interaction of the protein with peptide together with calcium ions was detected successfully, despite the relatively low molecular mass of the 2-kDa peptide. In addition, the attachment of smooth muscle cells to various surfaces has been monitored by TSM. These surfaces include laminin, fibronectin and bare gold. The results of these experiments in terms of changes of frequency (fs) and resistance (Rm) were analyzed. The responses of the surface-bound cells to the introduction of various ions, depolarisation events and damage subsequent to exposure to hydrogen peroxide were also observed. Morphological changes in the cells, as confirmed by atomic force microscopy and scanning electron microscopy, are correlated with results from the TSM sensor.

Passive air samplers (PAS) using XAD-resin were deployed at a wide variety of sites around the world for four years to asses the spatial and temporal trends of legacy organochlorine pesticides (OCPs) and current use pesticides (CUPs) in the global atmosphere. Legacy OCPs are prevalent in developing countries, whereas certain CUPs dominate in North America and Europe. OCP levels are declining world wide. Concentrations from the XAD-based PAS agreed with those from polyurethane foam (PUF) disk PAS within a factor of 4 for most sites. The comparison revealed that the sampling rate of the PUF-based PAS is more dependent on wind speed, whereas that of the XAD-PAS has a higher dependence on temperature. Analysis of PAS deployed across arid, subtropical Botswana showed that recent use has more impact on present day air concentrations than historical use. Year-long measurements by high volume air sampling in Botswana yielded higher HCB levels in winter, and higher endosulfan levels in summer. Those variations are neither related to temperature fluctuation nor seasonal hydrological events, and are therefore more likely caused by pesticide usage pattern. Very low levels of OCPs were found in the warm, dry, low organic matter soils of Botswana, including in soils from historical use areas. Such soils appear to have a low long-term storage capacity for pesticides allowing for rapid volatilization. They are thus not long-term sources of pesticides to the atmosphere. Endosulfan sulfate levels were observed to increase in tropical soils with increasing elevation. Water samples from high altitude cloud forests in Costa Rica contained very low concentrations of CUPs and it is unlikely that that those levels pose a threat to amphibians in cloud forests. Laboratory experiments and literature analysis showed that the inert gas stripping method for the determination of air-water partitioning equilibria is susceptible to surface sorption artifacts for chemicals with a interface-air partition coefficient log (KIA/m) > -3. Using larger bubbles reduces the surface-to-volume ratio and produces accurate air-water partition coefficients for chemicals with log (KIA/m) < -1.2.

An important research area in medicine is molecular diagnostics of cancers and infectious diseases, which can be diagnosed, managed and treated more effectively with genetic information. We have developed an integrated sample to answer bacterial detection platform combining a simple, universal bacterial lysis approach and sensitive nanomaterial electrochemical biosensors. Lysis is rapid and effective at releasing intercellular nucleic acid targets. The platform was directly challenged with unpurified lysates and successful at determining the presence of clinically relevant concentrations within 30min from sample to answer. Another important aspect of biosensor development is the development of cheap and efficient methods for manufacturing nanostructured microelectrodes. Previously, we have used costly silicon wafers for fabrication. Here we explored alternate inexpensive materials for fabrication including printed circuit boards, plastics and glass. We show that plain borosilicate glass is effective for templated bottom-up fabrication, with comparable performance to expensive silicon based nanostructured microelectrodes. Current state-of-the-art readout of many biomarkers is hampered by serially addressing arrays of low cost biosensors, without the use of high cost active electronics. Here we have developed a new concept, solution-based electrochemical circuits, which makes highly multiplexed sensing feasible on the surface of low-cost, glass chips. This method utilizes the idea that physical separation of liquid on an insulator can result in electrochemical isolation. Using this we can reduce the number of outputs to 2&radic;n, where n would be the number of serially connected sensors. We use urinary tract infections as a model system and prove that we can accurately detect species and antimicrobial resistance in multiplexed formats at clinically relevant concentrations.

Pharmaceuticals, personal care products and dissolved organic matter (OM) are introduced to soil via irrigation with reclaimed wastewater. This thesis examines the basic factors that influence sorption of these components in soil. Sorption of dissolved OM samples of varying composition to clay surfaces was examined. Results indicate that preferential sorption is dependent on clay type but not necessarily OM composition. Analysis of soils revealed aliphatic components, carbohydrates and amino acids are prevalent at the soil-water interface whereas aromatics are inaccessible at the soil-water interface. No clear relationship between sorption affinity of 17β-estradiol, sulfamethoxazole, carbamazepine and phenanthrene and soil OM aromaticity or aliphaticity was observed. A negative relationship between sorption and O-alkyl content may be due to these components blocking contaminant access to high affinity sorption sites. Therefore, application of reclaimed wastewater to soils with O-alkyl-rich OM may result in higher mobility of contaminants.

Pharmaceuticals, personal care products and dissolved organic matter (OM) are introduced to soil via irrigation with reclaimed wastewater. This thesis examines the basic factors that influence sorption of these components in soil. Sorption of dissolved OM samples of varying composition to clay surfaces was examined. Results indicate that preferential sorption is dependent on clay type but not necessarily OM composition. Analysis of soils revealed aliphatic components, carbohydrates and amino acids are prevalent at the soil-water interface whereas aromatics are inaccessible at the soil-water interface. No clear relationship between sorption affinity of 17β-estradiol, sulfamethoxazole, carbamazepine and phenanthrene and soil OM aromaticity or aliphaticity was observed. A negative relationship between sorption and O-alkyl content may be due to these components blocking contaminant access to high affinity sorption sites. Therefore, application of reclaimed wastewater to soils with O-alkyl-rich OM may result in higher mobility of contaminants.

An important research area in medicine is molecular diagnostics of cancers and infectious diseases, which can be diagnosed, managed and treated more effectively with genetic information. We have developed an integrated sample to answer bacterial detection platform combining a simple, universal bacterial lysis approach and sensitive nanomaterial electrochemical biosensors. Lysis is rapid and effective at releasing intercellular nucleic acid targets. The platform was directly challenged with unpurified lysates and successful at determining the presence of clinically relevant concentrations within 30min from sample to answer. Another important aspect of biosensor development is the development of cheap and efficient methods for manufacturing nanostructured microelectrodes. Previously, we have used costly silicon wafers for fabrication. Here we explored alternate inexpensive materials for fabrication including printed circuit boards, plastics and glass. We show that plain borosilicate glass is effective for templated bottom-up fabrication, with comparable performance to expensive silicon based nanostructured microelectrodes. Current state-of-the-art readout of many biomarkers is hampered by serially addressing arrays of low cost biosensors, without the use of high cost active electronics. Here we have developed a new concept, solution-based electrochemical circuits, which makes highly multiplexed sensing feasible on the surface of low-cost, glass chips. This method utilizes the idea that physical separation of liquid on an insulator can result in electrochemical isolation. Using this we can reduce the number of outputs to 2&radic;n, where n would be the number of serially connected sensors. We use urinary tract infections as a model system and prove that we can accurately detect species and antimicrobial resistance in multiplexed formats at clinically relevant concentrations.

Little is currently known regarding the nature and consequences of interactions between photoactive surfaces, including mineral dust and ‘urban film’, and gas-phase pollutants in urban environments. In order to address this knowledge gap, this thesis explores the photochemical reactivity of these environmental surfaces in controlled laboratory settings. The photoenhanced ozonation of pyrene, a toxic product of incomplete combustion, proceeds at different rates and via different mechanisms at three model ‘urban film’ surfaces. These results are important because they suggest that the reactivity of a molecule on simplified surfaces may not accurately reflect its reactivity in the real environment. The photooxidation of isopropanol at the surface of TiO2, here used as a proxy for the photoactive component of mineral dust, yields gas-phase acetone. This chemistry is amplified by nitrate, a major surficial component of atmospherically processed dust. These results suggest that dust has the potential to convert non-absorbing species to photochemically active species, and thereby serve as a source of reactive organic radicals for further gas- or surface-phase chemistry. Oxalic acid, the most atmospherically abundant dicarboxylic acid, is efficiently oxidized to gas-phase CO2 at the surface of Mauritanian sand and Icelandic volcanic ash. These experiments indicate that the lifetime of oxalic acid may be limited in arid regions by Fe and Ti-catalyzed aerosol-phase photochemistry. Fluorotelomer alcohols (FTOHs), a class of industrial chemicals used in the production of surface coatings, undergo photooxidation at the surface of sand and ash to yield toxic and persistent perfluorinated carboxylic acids (PFCAs). These results provide the first evidence that the metal-catalyzed heterogeneous oxidation of FTOHs may act as a local source of aerosol-phase PFCAs. Illumination of Nigerien sand in the presence of gas-phase SO2 leads to the formation of surface-sorbed sulfate. This chemistry proceeds more efficiently on fine sand than on coarse sand. In chamber experiments, the illumination of SO2 in the presence of realistically produced dust aerosol results in new particle formation. Together, these results suggest that SO2 photochemistry at the dust surface has the potential to change not only dust hygroscopicity but also the net scattering potential of dust-containing air masses.