This study was conducted to investigate the urban geochemistry of the city of Windsor (Ontario) and to provide added source apportionment information to work being carried out by the Canadian government. The goal of this study was to investigate the distribution, spatial variation and sources of manganese in urban Windsor soil. The literature indicates that human exposure to high levels of manganese, via inhalation, can cause respiratory and/or neurological effects. At the outset of the present study it was first hypothesized that vehicular traffic was the dominant source of anthropogenic manganese. An alternative hypothesis was that there were multiple anthropogenic sources of manganese in Windsor. The sample collection scheme was designed to determine (1) the current and background soil concentrations of manganese in Windsor, (2) the spatial distribution of manganese in order to reveal sources of manganese, and (3) the manganese content of moss-sequestered airborne particles, which can potentially deposit onto the soil surface, using low-technology biomonitoring. The first phase of the study consisted of a preliminary soil survey which identified elevated areas of soil manganese concentrations. During this survey, the field efficiency of a field portable X-ray fluorescence (FPXRF) instrument, as well as sample preparation methods were evaluated. Efficiency of the FPXRF was determined by comparison to ICP-MS, a traditional trace element analysis method. The preliminary soil survey identified several areas of elevated (ranging from 884 to 2390 ppm) soil manganese which were further investigated during the second, more complete, soil survey. The moss biomonitoring technique of using moss bags was used to collect airborne particles for semi-quantitative analysis. Analysis of soil samples included total manganese and other trace elements, pH, moisture and carbon content, and manganese speciation. Urban Windsor soil manganese distribution revealed both natural and anthropogenic sources of soil manganese and three distinct soil sample types, transect, baseline and natural. In general, manganese in Windsor had a west-to-east trend of decreasing levels in soil and moss-sequestered airborne particles. The latter showed a modern-day elemental signature while the former (collocated soil) a legacy elemental signature. It was concluded that both the FPXRF instrument and the moss biomonitoring technique can be useful screening tools in studies of urban environments.

Friedreichs ataxia (FRDA) is a progressive spinocerebellar ataxia (SCA) inherited as an autosomal recessive trait. The neurodegeneration, cardiomyopathy, diabetes mellitus and skeletal deformities characteristic to FRDA result from a deficiency in the mitochondrial protein frataxin. Frataxin chaperones iron to heme and iron-sulfur clusters and its deficiency causes mitochondrial iron accumulation and oxidative stress.<p> To address the effect of frataxin deficiency on mitochondrial iron chemistry, mitochondria were isolated from FRDA and control fibroblasts. X-ray absorption spectroscopy showed that ferrihydrite was the predominant form of iron in both. Near edge analysis showed that the ferrihydrite in the FRDA mitochondria resembled the highly organized ferrihydrite of ferritin. Western blotting confirmed that FRDA mitochondria had 3-fold more holoferritin containing stainable iron. I conclude that mitochondria from FRDA fibroblasts mineralize excess iron as ferrihydrite within mitochondrial ferritin.<p> To address how cellular iron dysregulation affected metal distribution in brain and spinal cord, a new synchrotron imaging technique, rapid-scanning x-ray fluorescence (RS-XRF) was employed and validated. Brain structures were readily identified by their unique metal content and distribution. This showed that RS-XRF could be used to reveal metal pathologies associated with diseases of metal metabolism such as FRDA. Since human FRDA tissues were not available for a detailed study, RS-XRF was employed to study the distribution of metals in normal cerebellum, a major site of FRDA-associated neurodegeneration, and to localize and quantify metals in the brain and spinal cord from a patient with a SCA of unknown aetiology. The motivation for this work is the prospect of future systematic studies on metal pathology in neurodegenerative diseases with direct application to FRDA. Novel findings arising from this work were the metal segmentation of the dentate nucleus, the high copper content of the olivary region and the different metal content of lesions at different stages of neurodegeneration. My results suggest that not only iron, but also copper and zinc may play a role in the physiopathology of neurodegeneration. Therefore, all three metals should be investigated in FRDA and other SCA of both known and unknown aetiologies to identify possible new therapeutic targets.

The primary aim of the work presented in this thesis is to elucidate novel information regarding the uptake, storage, distributions, and functions of both copper and zinc in mammalian cells by predominantly using a combination of the high resolution imaging modalities, synchrotron radiation X-ray fluorescence microscopy (SXRF) and standard fluorescence imaging. Results from studies using cell permeable, metal ion selective fluorescent probes suggested the presence of labile pools of copper and zinc localized within the mitochondria and Golgi apparatus. Furthermore, SXRF imaging of a cell line defective in the copper transporter, Atox1, revealed intriguing differences in the Cu distribution of Atox1-/- cells compared to the corresponding wild-type cells. Finally, spatially well-resolved SXRF elemental maps of single, adherent mouse cells revealed remarkable changes in the distributions of both zinc and copper as the cells progressed through the cell cycle. Taken together, findings suggested major roles for copper and zinc within a native biological setting.

This research developed an on-line measurement systemfor determining the amount of fluorochemicals on carpet fibers using energy-dispersive X-ray fluorescence (EDXRF).This system is designed as a complementary tool to an existingchemical burn test certified by the American Association ofTextile Chemists and Colorists (AATCC), which is performed off-line on randomly selected carpet samples and time consuming.This research reviewed XRF principles and determined parameters that affect XRF spectra such as measurement time, measurement number, X-ray tube voltage, X-ray tube current, primary beam filter, and carpet characteristics. For this application, XRF calibrations must be performed for carpets of all styles and types. However, preparing actual carpet calibration samples is expensive. This research introduced a methodology to synthetically generate reference spectra using XRF spectra from standard fluorochemical stock solution samples and from base carpet samples for each carpet type to be tested. Thus, actual, physical standards are not required for each carpet type or style. This study showed that the synthetically generated XRF spectra alone were not always sufficient to guarantee the confidence interval required by the certified AATCC test. Thus, it is recommended that for on-line implementation, burn test results should be used to create a historical data base for each carpet type to reduce margin of error for calibrations generated from the synthetic spectra.

Mercury (Hg) compounds are some of the most toxic compounds of any heavy metal on earth. Due to long-range transport from point sources Hg can be found world-wide in air, soil, water, and living organisms. Mercury compounds can cause a number of adverse effects, with the unborn fetus, infants, and children being most susceptible. Zebrafish (Danio rerio) are an excellent vertebrate model system for toxicological studies, including developmental effects. The overall objective of this research was to investigate the effects of inorganic forms of Hg in zebrafish larvae. Unique accumulation patterns were observed using synchrotron X-ray fluorescence imaging after zebrafish were exposed to one of four Hg compounds (i.e. mercuric chloride, mercury bis-L-cysteineate, methylmercury chloride, methylmercury L-cysteineate). Specifically, we noted chemical form dependant and tissue-specific Hg accumulation including the sensory cells of the olfactory epithelia and the neuromasts. Phenylthiourea (PTU) is commonly used to block zebrafish melanogenesis to generate transparent larvae to aid with enhanced visualization of immunohistochemical and vital stains. It was determined that PTU dramatically alters Hg toxicity through chemical interaction with Hg so that further studies were conducted in the absence of PTU. To investigate the effects of Hg on primary neurons, the immunohistochemistry protocol using anti-acetylated tubulin was performed and the results demonstrated that mercuric chloride damages primary neurons particularly in the olfactory pits. To study potential detoxification of Hg in zebrafish we examined the efficacy of two sequestration agents, dimercaptosuccinic acid and alpha lipoic acid, as well as endogenous selenium. The levels of Hg were not significantly lower following treatment with either sequestration agent under the conditions used in this research. Previous work examining the antagonistic relationship between Hg and selenium has been conducted by dosing animals with both Hg and selenium (Se). We discovered a mixed chalcogenide of the general form HgSxSe(1-x) forming in vivo following exposure to mercuric chloride without the addition of selenium. Indeed the selenium may have been remobilized from natural stores in the pigment spots. The research presented herein demonstrates that the target tissues for Hg depend strongly on chemical form. In particular inorganic Hg can accumulate in a number of important tissues including sensory systems. The formation of insoluble and non-toxic HgSxSe(1-x) in zebrafish larvae suggests that endogenous selenium may play critical roles in modulating toxicity.

The Neolithic and late Iron Age pottery from the settlement site of Pool, Sanday, Orkney, was studied on two levels. Firstly, a morphological and technological study was carried out to establish a sequence for the site. Secondly an assessment was made of the usefulness of X-ray Fluorescence Analysis, Inductively Coupled Plasma Spectrometry and Petrological analysis to coarse ware studies, using the Pool assemblage as a case study. Recording of technological and typological attributes allowed three phases of Neolithic pottery to be identified. The earliest phase included sherds of Unstan Ware. This phase was followed by an assemblage characterised by pottery with incised decoration, which was stratified below a traditional Grooved Ware assemblage. The change in pottery styles and manufacturing methods with the Grooved Ware indicated that it evolved elsewhere. Grass tempered and burnished pottery characterised the Iron Age assemblage. Pottery samples from all phases of the site were analysed by XRF and ICPS. In addition, pottery from late Iron Age sites in the area was analysed for comparison with the Pool Iron Age pottery. XRF and ICPS analyses did not distinguish between either different phases at Pool or different Orcadian sites. This was attributed to the similarities in geological deposits over much of Orkney and the variations which can occur within a clay source. A clay survey was carried out in the vicinity of the site, and samples taken for comparison with the Pool pottery. Identification of rocks and minerals in thin section, and grain-size analysis, indicated that the Pool pottery was made locally to the site, and that both primary and secondary clays were used. It was concluded that petrological analysis is more suitable than elemental analysis in the study of coarse wares.

This study was conducted to investigate the urban geochemistry of the city of Windsor (Ontario) and to provide added source apportionment information to work being carried out by the Canadian government. The goal of this study was to investigate the distribution, spatial variation and sources of manganese in urban Windsor soil. The literature indicates that human exposure to high levels of manganese, via inhalation, can cause respiratory and/or neurological effects. At the outset of the present study it was first hypothesized that vehicular traffic was the dominant source of anthropogenic manganese. An alternative hypothesis was that there were multiple anthropogenic sources of manganese in Windsor. The sample collection scheme was designed to determine (1) the current and background soil concentrations of manganese in Windsor, (2) the spatial distribution of manganese in order to reveal sources of manganese, and (3) the manganese content of moss-sequestered airborne particles, which can potentially deposit onto the soil surface, using low-technology biomonitoring. The first phase of the study consisted of a preliminary soil survey which identified elevated areas of soil manganese concentrations. During this survey, the field efficiency of a field portable X-ray fluorescence (FPXRF) instrument, as well as sample preparation methods were evaluated. Efficiency of the FPXRF was determined by comparison to ICP-MS, a traditional trace element analysis method. The preliminary soil survey identified several areas of elevated (ranging from 884 to 2390 ppm) soil manganese which were further investigated during the second, more complete, soil survey. The moss biomonitoring technique of using moss bags was used to collect airborne particles for semi-quantitative analysis. Analysis of soil samples included total manganese and other trace elements, pH, moisture and carbon content, and manganese speciation. Urban Windsor soil manganese distribution revealed both natural and anthropogenic sources of soil manganese and three distinct soil sample types, transect, baseline and natural. In general, manganese in Windsor had a west-to-east trend of decreasing levels in soil and moss-sequestered airborne particles. The latter showed a modern-day elemental signature while the former (collocated soil) a legacy elemental signature. It was concluded that both the FPXRF instrument and the moss biomonitoring technique can be useful screening tools in studies of urban environments.

The objective of this thesis is to categorize carpet types according to their XRF spectra and verify if further classification of carpets is possible for use of an XRF analysis system in the carpet manufacturing line. This thesis consists of (1) implementing and studying effective algorithms for automated analysis of X-ray spectra, (2) comparing known algorithms for X-ray spectra analysis, and (3) implementing our own algorithm for classification of carpets spectra obtained for further fluorine online analysis of XRF inspected carpets. This research is intended for quick and accurate automated analysis of raw XRF spectra and matching analysis results to a database of XRF spectra of raw carpets. The research uses spectrum signal processing and spectrum analysis regarding efficacy of combined methods for XRF inspected carpets. X-Ray Fluorescence is a key technology for detection of chemical elements. Fluorine is a key element for carpet's quality. XRF has been chosen to be a potential candidate to measure fluorine since it is a versatile tool for low concentration element detection. Due to specific XRF background spectrum for each different carpet type, carpet samples may need specific calibrations for further computation of carpet fluorine concentration. Automating the detection of the carpet type is intended to help in automating the XRF calibration.

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Metal ions play essential roles in biological functions including catalysis, protein stability, DNA-protein interactions and cell signaling. It is estimated that 30% of proteins utilize metals in some fashion. Additionally, methods by which metal ions can be visualized have been utilized to study metal concentrations and localizations in relation to disease. Understanding the roles metals play in biological systems has great potential in medicine and technology. Chapters 1 and 2 of this dissertation analyzes the structure and function of the Mn-dependent enzyme oxalate decarboxylase (OxDc) and Chapter 2 presents a bioinformatic analysis of the cupin superfamily that provides the structural scaffold of the decarboxylase. The X-ray crystal structure of the W132F variant was determined and utilized together with EPR data to develop a computational approach to determining EPR spectra of the enzyme’s two metal-binding centers. Furthermore, a variant in which the catalytic Glu162 was deleted revealed the binding mode of oxalate, the first substrate-bound structure of OxDc. OxDc is a member of the cupin superfamily, which comprises a wide variety of proteins and enzymes with great sequence and functional diversity. A bioinformatics analysis of the superfamily was performed to analyze how sequence variation determines function and metal utilization. Chapters 3 and 4 discuss the expansion of lanthanide-binding tags (LBTs) to in cellulo studies. Lanthanide-binding tags are short sequences of amino acids that have high affinity and selectivity for lanthanide ions. An EGF-LBT construct used to quantify EGF receptors on the surface of A431 and HeLa cells. The results from the LBT quantification are consistent with previous studies of EGFR receptors in these cell types, validating the use of this method for future studies. The potential of using LBTs for X-ray fluorescence microscopy (XFM) was also investigated. LBT-labeled constructs were utilized to investigate if membrane bound as well as cytosolic LBT-containing proteins could be visualized and localized to their cell compartments via XFM; both membrane-localized and cytosolic proteins were successfully visualized. With the high resolution (< 150 Å) obtainable with new synchrotron beamline configurations LBTs could be used to study nanoscale biological structures in their near-native state.