Há fortes evidências dos efeitos negativos da exposição gestacional a poluentes atmosféricos. No entanto, mecanismos de atuação de poluentes não são bem compreendidos. Alterações fisiológicas anômalas na progenitora, durante o período de gravidez, podem causar mudanças permanentes na prole, que podem desencadear futuras doenças na vida adulta. Portanto, o estudo dessas alterações maternas é importante. A metabolômica é definida como a análise global do metaboloma de um organismo em experimentos comparativos, com o objetivo de observar mudanças relativas da abundância dos metabólitos, o aparecimento ou desaparecimento de metabólitos, e pode fornecer uma melhor compreensão dos mecanismos de funcionamento celular dos organismos a nível molecular. Nesse trabalho, um estudo experimental de exposição gestacional materna, ao material particulado fino (MP2,5), foi realizado, para avaliar os efeitos dessa exposição no metabolismo, por meio da análise metabolômica global da urina de camundongos fêmeas progenitoras expostas ao MP2,5 (grupo teste) ou a ar filtrado (grupo controle) durante a gestação. Um método cromatográfico e de preparo de amostra para metabolômica urinária por HILIC-MS foram otimizados. Para a otimização da condição cromatográfica, foram investigados a influência de aditivos, concentração de sal e pH da fase móvel, bem como, a rampa do gradiente. A melhor condição foi escolhida por meio da avaliação do formato de pico, da intensidade relativa e do CV do tempo de retenção para 15 m/z selecionados, assim como, pelo número total de molecular features e CV da intensidade desses molecular features. A melhor condição obtida apresenta 20 mmol/L de formiato de amônio em sua composição do solvente B da fase móvel e 95% acetonitrila e 5% solução aquosa 400 mmol/L de formiato de amônio na composição do solvente A. Para o preparo de amostra, foram examinados diferentes solventes orgânicos e suas misturas, para a precipitação de proteínas da urina. O isopropanol foi o solvente apresentou os melhores resultados para o preparo de amostra. Dessa forma, com o método analítico otimizado, as amostras de urina de camundongos fêmeas prenhas foram submetidas à analise metabolômica global por HILIC-MS. O metaboloma dos animais foi bastante alterado pela exposição gestacional ao material particulado. Observou-se alteração dos níveis de carnitinas, aminoácidos, peptídeos, entre outros. Há também indícios de que a poluição atmosférica alterou a microbiota intestinal dos animais, devido ao aumento de N-óxido de trimetilamina, um metabólito que também é relacionado ao processo de aterosclerose. Níveis de metabólitos relacionados ao metabolismo da histidina também foram alterados devido a exposição ao MP2,5. Níveis de carnitina e acilcarnitinas foram aumentados no teste, sugerindo alteração da produção de energia na mitocontria. / There are strong evidences regarding negative effects of gestational exposure to air pollution. However, the mechanisms of action of air pollutants are not well established. Maternal anomalous physiological changes during pregnancy may cause permanent changes in offsprings, that might initiate future diseases in adult life. Therefore, the study of those maternal changes during pregnancy is important. Metabolomics is defined as the global analysis of the metabolome of an organism in comparative studies, for the measurement of relative changes in the metabolite abundance, appearance or disappearance. Metabolomics might provide a better understanding of cellular functioning at the molecular level. In this work, an experimental study of maternal gestational exposure to fine particulate matter (PM2.5) was accomplished to evaluate the effects of this exposure to the metabolism, by an untargeted metabolomics analysis of urine from pregnant mice exposed to PM2.5 or to filtered air during pregnancy. A chromatographic and sample preparation methods for urinary untargeted metabolomics analysis by HILIC-MS were optimized. For the chromatography optimization, the influence of mobile phase additives, salt concentration and pH, as well as, the gradient ramp were investigated. The best condition was chosen by the evaluation of peak shape, relative intensity and retention time CV of 15 selected m/z, as well as, the total number of molecular features and the intensity CV of those molecular features. The best condition comprises of 20 mmol/L of ammonium formate as solvent B, and 95% acetonitrile and 5% 400 mmol/L of ammonium formate as solvent A, in the composition of the mobile phase. For the sample preparation, different solvents, along with, their mixtures were examined for the urine protein precipitation. Isopropanol was the solvent that presented the best results for sample preparation. Thus, after the analytical method optimized, urine samples from the progenitors were submitted to untargeted metabolomics analysis by HILIC-MS. The animals\' metabolome were significantly changed by the gestational exposure to particulate matter. It was observed changes in the levels of carnitines, amino acids, peptides, among others. There is some indication that the air pollution has altered the gut microbiota, due to the enhancement of trimethylamine N-oxide (TMAO), a metabolite that is also related to the atherosclerosis process. The level of metabolites related to histidine metabolism were also altered due to PM2.5 exposure. Carnitine and acylcarnitine levels were also increased in the test group, suggesting an altered energy production in the mitochondria.

Ambient air pollution, including airborne particulate matter (PM) derived from combustion of fossil fuels (FF) or biomass (BM), has detrimental inflammatory effects on human health. Myeloid antigen presenting cells, including dendritic cells (DCs) regulate immune responses in the airway and sample inhaled PM. This study tests the hypothesis that PM interacts with multiple environmental sensing pathways in DCs with outcomes that depend on particle size and composition as determined by combustion source. The effects of different sized PM (< 10μm, PM10; < 2.5μm, PM2.5), derived from the combustion of FF or BM, on human monocyte-derived or ex vivo sputum DCs, were examined. DC activation status, cytokine production and aryl hydrocarbon receptor (AhR) signalling were assessed by flow-cytometry, multiplex ELISA and qRT-PCR, following exposure to PM. Pathway-specific antagonists were used to explore underlying mechanisms. Particle size and combustion source influenced the effects of PM on DCs. Irrespective of combustion source, PM10 but not PM2.5, induced MoDC maturation and stimulated production of inflammatory cytokines, including IL-1β and IL-18, indicative of inflammasome activation. These responses were dependent, at least in part, on TLR4 as was the induction of IDO by PM10. AhR signalling was induced by PM in both MoDC and ex vivo sputum DC. It was stimulated by both PM10 and PM2.5 and was induced more strongly by BM-derived PM. AhR activation was independent of DC maturation and TLR4 signalling. Additionally, BM- but not FF-derived PM increased NADH levels in DC suggestive of altered metabolism. Thus, PM induces a complex programme of DC activation, influenced by size and combustion source, which includes classical maturation, inflammasome dependent cytokine release and AhR signalling as well as potential metabolic changes. In the airway, exposure to different PM and the changes in DCs induced by them may lead to altered responses to inhaled antigen.

>Magister Scientiae - MSc / Ambient air pollution is the biggest environmental threat to human health. According to the World Health Organisation (WHO), ambient air pollution kills millions of people worldwide every year. Airborne particulate matter (APM) affects more people than any other air pollutant and has been linked with various adverse health outcomes, especially fine fractions (commonly abbreviated to PM2.5). PM2.5 penetrates lung tissue to enter the cardiovascular system where it poses the greatest risk. Detailed ambient APM studies are rare in Africa. Such studies are needed to better understand the characteristics, origins and trends of particulate pollution. This study was conducted in Cape Town (the first of its kind for the area) as part of a bigger project on ambient PM2.5 and soot concentrations in South Africa. PM2.5 filter samples were collected at a fixed sampling site in the suburb of Kraaifontein from April 2017 to April 2018, yielding 121 days of data. PM2.5 mass concentration and absorption coefficient determinations were done using gravimetric analysis and smoke stain reflectometry (SSR). Mean PM2.5 concentration for the study period was 13.4 ± 8.1 μg.m-3 (range: 1.17-39.1 μg.m-3) that fell below the South African National Ambient Air Quality Standard (SA NAAQS) annual limit of 20 μg.m-3 but exceeded the WHO annual limit of 10 μg.m-3. Mean absorption coefficient for the same period was 1.38 ± 1.23 m-1.10-5 (range: 0.00-5.38 m-1.10-5) which did not exceeded any limits. Source-region analyses were performed using single, 24-hour backward trajectories and trajectory clusters derived from the Hybrid Single Particle Lagrangian Integrated Trajectory (HYSPLIT) model. Six single trajectories were identified; the most frequent were trajectories Atlantic-Ocean (38.8 %) and Indian-Ocean (26.4 %). Cluster analyses yielded three to four clusters per season. Dominating clusters were Atlantic-Ocean (61.8 %) and Indian-Ocean (29.5 %) and Inland (8.50 %). Contributions by local sources (within 40 km of the sampling site) to PM mass in samples far exceeded those of distant sources through long-range transport (LRT).

The body of work in this dissertation focuses on the properties of an environmental thin film system, including the roughness and composition of the surface. The deposition of particles, such as airborne soil and plant pollen, from the atmosphere creates a thin film known as “environmental film” or “urban film” that covers virtually all of Earth’s solid surfaces. Environmental films have been shown to accumulate a variety of chemicals, including toxic pollutants. To investigate the means by which environmental films uptake chemicals, model films are made in the lab and real films are collected outside. Model films serve to mimic the properties of native films and allow for a simple analysis of a complex system. Native films serve to provide real field samples to analyze. The properties of model and native films are characterized using reflected light to determine what the film is made of and microscopes capable of imaging small particles. The results of the model film study indicate a model capable of reproducing the surface roughness and other properties of native films. This study serves as a platform with the goal of making model films that better mimic native films. The results of the native film study indicate successful imaging using microscopes capable of revealing the structure and chemical composition of the films. This imaging adds an important contribution to the field that has not previously been performed.

Airborne particulate matter (PM) consists of solid and liquid particles suspended in air. PM causes many negative health effects when inhaled like exacerbations of asthma, chronic obstructive pulmonary disease (COPD), and premature death. The health impacts of PM depend on the physical size and chemical composition of the inhaled particles. Particles less than 2.5 micrometers can penetrate the deep lung and enter the bloodstream. Understanding the composition of PM helps study human exposures and evaluate PM sources to support control and mitigation strategies. This thesis examines PM in power plant emissions, in ambient air, and in homes, with an emphasis on characterizing hazardous metals. PM emissions from the University of Iowa power plant were examined during a transition away from coal. Biomass is a renewable fuel and when used in place of or co-fired alongside coal it directly reduces emissions of fossil CO2 and PM to the atmosphere. PM emissions were examined under two scenarios: the first was a stoker boiler that fired 100% renewable energy pellets. PM and metal emissions reduced by 59 and 80% respectively despite increases to polycyclic aromatic hydrocarbons (PAHs) that was attributed to emission of unburned carbon fuel. The second test was done in a circulating fluidized bed boiler firing 78% oat hulls, 17% coal, and 5% energy pellets. Decreases in PM, PAHs, and metal emissions were 32, 33, and 50%, respectively. Reductions in PM, PAHs, and metals when firing renewable fuels provide environmental advantages to local air quality while trying to eliminate the use of coal. Ambient air quality was examined downwind of the power plant. The levels of PM2.5 in Iowa City in 2016 ranged from 1.4-32.1 µg m-3 with an annual mean of 7.5 µg m-3. These levels are below the National Ambient Air Quality Standards (NAAQS) for 24-hours (35 µg m-3) and the annual average (12 µg m-3). On average, the analyzed metals accounted for 10.7 ± 5.3% of PM2.5. Potassium, calcium, and zinc were the most abundant metals (averaging 250 ± 10, 170 ± 40, and 11.2 ± 0.5 ng m-3, respectively) and are associated with geological sources and biomass burning. Metals associated with fossil fuel combustion such as arsenic, lead, or vanadium were observed at low levels (averaging 0.58 ± 0.01, 1.32 ± 0.03, and 0.19 ± 0.01 ng m 3, respectively). Harmful metals were at least an order of magnitude lower than the World Health Organization’s guideline concentrations in Iowa City PM2.5. Overall, the hazardous metals in ambient PM2.5 observed in Iowa City were not at concerning levels. The work in this chapter helps provide a metal speciation profile for future studies and these measurements can be used to assess future changes in PM2.5 metal concentrations. Indoor PM collected in homes of Eastern Iowa COPD patients was characterized for its metal content. COPD patients are at higher risk of developing respiratory infections, which cause acute exacerbations of COPD—the leading cause of mortality in COPD patients—and airborne PM increases risk of infection. From the 21 homes studied 6-87 mg of indoor PM was collected over 30 days in winter. Crustal metals such as magnesium, iron, and aluminum were the most concentrated in indoor PM, having mass fractions from 3000-25,000 ng mg-1. Toxic metals like vanadium and arsenic were at lower levels from 3-65 ng mg-1. Between homes the relative abundance of metals was similar, but the absolute abundance was highly variable. Analyzing indoor PM is essential since Americans on average spend 80% of their day indoors making it a significant environment for PM exposure. This chapter provides a chemical profile for different homes when studying the impact PM has on respiratory health. This thesis also provides new measurements of the chemical composition of PM at the point of emission, in ambient air, and within homes. Advancing knowledge of PM composition in different environments is vital in understanding its impacts on human health. Improvements to local air quality with reductions to PM and metal emissions were observed when firing alternative fuels. Ambient PM2.5 concentrations in Iowa City were lower than NAAQS levels and the composition of both ambient and indoor PM was evaluated. With greater understanding of PM composition, better control strategies can be studied and employed to improve local air quality.

During wintertime temperature inversion episodes the concentrations of particulate air pollution, also defined as particulate matter (PM), in Utah’s Cache Valley have often been highest in the nation, with concentrations surpassing more populated and industrial areas. This has attracted much local and national attention to the area and its pollution. The Cache Valley has recently been declared to be in non-attainment of provisions of Federal law bringing to bear Federal regulatory attention as well. While there is epidemiological evidence indicating that PM is detrimental to public health, there is much less information indicating by which biological and molecular mechanisms PM can exert harm. This study was undertaken to better understand the mechanisms by which ambient PM collected in the Cache Valley can be harmful to human lung cells. Cache Valley PM was found to be mildly cytotoxic only at concentrations that were much greater than physiologically achievable, and such concentrations were difficult to obtain with the limited amounts of captured ambient PM. The limited cytotoxicity was despite apparent PM-induced pro-apoptotic signaling such as caspase-3 upregulation, and activation of caspase-12 and calpain. Cache Valley PM was found to be stressful to cells, triggering endoplasmic reticulum stress and the unfolded protein response. Cache Valley PM was also found to be inflammogenic leading to activation of pro-inflammatory transcription factors, increases in the release of pro-inflammatory cytokines and chemokines, as well as the upregulation of the activating receptors of these cytokines. The proinflammatory effects and absence of apoptosis, despite pro-apoptotic signaling of the Cache Valley PM on human lung cells appeared to stem from increased activation of the central pro-growth protein Akt with subsequent inactivation of the tumor suppressor P-TEN. These findings have indicated novel mechanisms of PM-related cellular stress and inflammation contributing needed information on what may be underlying mechanisms of PM associcated illnesses.

The lungs represent one of the earliest interfaces for pathogens and noxious stimuli to interact with the body. As such, careful maintenance of the permeability barrier is vital in providing homeostasis within the lung. Essential to maintaining this barrier is the tight junction, which primarily acts as a paracellular seal and regulator of ionic transport, but also contributes to establishing cell polarity, cell-to-cell integrity, and regulating cell proliferation and differentiation. The loss of these tight junctions has been documented to result in alterations in inflammation, and ultimately the development of many respiratory disorders such as COPD, Asthma, ARDS, and pulmonary fibrosis. One critical contributor that creates this permeability barrier is the tight junctional protein Claudin. While studies have begun to elucidate the various functions and roles of various Claudins, our understanding is still limited. To initially investigate these proteins, we looked at both temporal and spatial expression patterns for family members during development. A consistent pattern was demonstrated in mRNA expression for the majority of Claudin members. In general, Claudin expression underwent rapid increase during time periods that correlate with the pseudoglanduar/canalicular periods. One notable exception was Claudin 6 (Cldn6), which demonstrated decreasing levels of mRNA expression throughout gestation. We also sought to understand expression dynamics during the addition of maternal secondhand smoke (SHS) which resulted in an almost universal decrease in Claudin proteins. To more fully explore expression mechanisms that affect Claudin-6 (Cldn6), we exposed pulmonary alveolar type II (A549) cells to cigarette smoke extract (CSE) and found that it transcriptionally regulated Cldn6 expression. Using a luciferase reporter, we determined that transcription was negatively regulated at multiple promoter response elements by CSE, and transcription was equally hindered by hypoxic conditions. These findings identified Cldn6 as a potential target of SHS and other respiratory irritants such as diesel particulate matter (DPM). We next sought to assess whether an increase in Cldn6 was sufficient to provide a protective advantage against harmful exogenous exposure. To test this, we utilized a doxycycline induced Cldn6 over-expressing mouse, and subjected it to SHS for 30 days to stimulate an inflammatory state. Our findings demonstrated that Cldn6 transgenic animals have decreased inflammation as evidence by decreased total cell infiltration into the airways, decreased polymorphonuclocyte (PMNs) extravasation, total protein in bronchoalveolar lavage fluid (BALF), and decreased cytokine secretion. Anti-inflammatory advantages were also discovered during experiments involving acute exposure to DPM. In both cases, while stimulation of transgenic mice with SHS or DPM diminished Cldn6 expression, anti-inflammatory evidence emerged suggesting that genetic up-regulation of Cldn6 likely causes the recruitment of other tight junctional components during an organism's response to environmental assault.

This thesis concerns various aspects of the influence of atmospheric particulate matter on the terrestrial system. In Part I, the radiative influence of particulate matter on the production of crops is explored using the CERES crop model. It is known that the presence of aerosols in the atmosphere simultaneously reduces the amount of sunlight reaching the surface and increases the fraction of that light which is diffuse. Reduction of the total amount of sunlight tends to slow the rate of photosynthesis occurring in plants while increasing the fraction which is diffuse tends to increase the net rate of photosynthesis by more evenly distributing sunlight throughout all layers of a plant canopy. The CERES crop model was modified to estimate the influence of both a reduction in total sunlight and an increase in the diffuse fraction. Model simulations were performed for rice, maize and wheat at a variety of locations and found the likely influence of aerosols on crop production to be a 0-10% reduction in yield compared to the base case. In Part II, the concentration and size distribution of water-insoluble aerosols (WIA) is explored. It has been well-established that atmospheric particulate matter influences the planetary radiation budget both directly and indirectly. The magnitude of these influences is related to particle solubility. A new technique was developed to provide these measurements in real-time. This instrumentation was evaluated in a laboratory setting and implemented into several field studies. Results from these sampling campaigns indicate that in areas heavily influenced by motor vehicle traffic, the WIA concentration is dominated by particulate soot. Many episodes of high levels of crustal dust were also observed. At these times, the WIA size distribution shifted toward larger sizes in a characteristic manner. This suggests that this method is useful for detecting insoluble mineral aerosols as well as particulate soot and that examination of the WIA size-distribution may provide a basis to distinguish between the two.

Water samples were collected along salinity gradients during different seasons from three estuaries (Tseng-Wen, Gao-Ping, Er-Ren) and coasts in/off southwestern Taiwan. In order to assess the partitioning of trace metals between solution and particle, the concentrations of dissolve and particulate trace metals (Cd, Cu, Ni, Pb) along with their chemical affinity fractions, were determined. This study investigated the variations in distribution and partitioning of the different metals in estuarine waters, and examined the effects of oxides and particulate organic matter on the partitioning of trace metals in waters from different estuaries. Fractionation of dissolved trace metal species was based on ion exchange (Chelex-100 and AG MP-1 resins) separation techniques. The fractions obtained were operationally defined as labile (Chelex), organic (AG MP-1) and inert. Particles were extracted to three phases (surface adsorbed phases, Fe¡VMn oxide/organic phases and refractory phases) using sequential extraction techniques. Seasonally variable distributions of dissolved trace metals were found in the Tseng-Wen estuary. The behavior of trace metals was mainly influenced by anthropogenic input during the dry season in the upper Tseng-Wen estuary, while mixing processes controlled the distribution of trace metals during the wet season. The dilution effect was a major factor in the metal distribution in the Gao-Ping estuary due to high river discharge. The higher concentration of metals in the Er-Ren estuary, in contrast to other estuaries indicated that the Er-Ren estuary has serious pollution concerns. According to the results of particulate metal fractions obtained, Cd and Pb existed predominantly in the surface adsorbed phase. The speciation and spatial distribution of Cd were similar to those of Mn, indicating that the formation of authigenic Mn oxides affected the distribution of Cd in estuaries. The percentage of oxide /organic phase for Cu accounted for 25% of total particulate Cu, but dose not correlate well with particulate organic carbon, implying that organic carbon is not the only factor controlling particulate Cu distribution. Ni was present mainly in lattice phase, except in the Er-Ren estuary where anthropogenic Ni loading was high. In the Tseng-Wen and Gao-Ping estuaries, the percentages of lattice phase of all metals determined in this study during the wet season were higher than those during the dry season. These seasonal variations are probably resulted from different flushing times in dry/wet seasons, which control the extent of geochemical processes for trace elements.

The Tapered Element Oscillating Microbalance (TEOM) sampler is an EPA approved Federal Equivalent Method Sampler for measuring PM10 concentrations. The Center for Agricultural Air Quality Engineering and Science (CAAQES) owns two Rupprecht and Patashnick (R&P) Series1400a monitors. The R&P Series 1400a monitor uses the TEOM method to measure particulate matter (PM) concentrations and was approved by EPA in 1990 as an automated equivalent method PM10 sampler. Since its approval, many state air pollution regulatory agencies (SAPRAs) have located R&P Series 1400a monitors at community-oriented monitoring sites. Some SAPRAs have even located TEOM samplers at the property line of major sources to determine if the source is meeting its permit requirements for PMc emissions. This thesis presents the results of PM10 and TSP concentrations measured with TEOM and low-volume gravimetric samplers at two Texas cattle feedlots. The purpose of this research was to compare the performance of the R&P Series 1400a monitor to the low-volume gravimetric sampler when sampling PM from a feedlot. Furthermore, this research was conducted to avoid the inappropriate regulation of cattle feedlots that may occur in the future as a consequence of the TEOM sampler being used to measure PMc emissions. The results of this research demonstrate that relationship between the R&P Series 1400a monitor and the low-volume gravimetric sampler is linear. In general, it was observed that the TEOM sampler measured higher PM10 and TSP concentrations than the low-volume gravimetric sampler when sampling downwind from a cattle feedlot. The opposite results were observed when sampling was conducted upwind from the feedlot. The collected data demonstrates that the concentration difference between the two sampling methods is linearly dependent with the concentration intensity for the upwind sampling locations. This trend was shown to be statistically significant. Another linear relationship was observed between the concentration difference and the particle size (mass median diameter and geometric standard deviation) of the sampled dust. Although this trend was not statistically significant, it is believed that additional downwind concentration measurements would validate this relationship.