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The purpose of the current study was to test the hypothesis that chronic hypoxia associated with sleep-disordered breathing relates to abnormal Nitric Oxide (NO) production and vascular endothelial growth factor (VEGF) expression patterns that contribute to aberrancy of specific determinates of cardiovascular and cardiopulmonary function before, during, and after graded exercise. These patterns may further reflect pathologic alteration of signaling within the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt-1) transduction network. To this end, 7 medically diagnosed OSA patients (3 male, 4 female), mean age 48 years and 7 apparently healthy control subjects (3 male, 4 female), mean age 42 years, underwent baseline venous blood draws and maximal bicycle ergometry. Mononuclear cells isolated from peripheral blood were utilized as reporter cells for measurement of VEGF, Akt-1, hypoxia inducible factor-1 alpha (HIF-1 alpha), and vascular endothelial growth factor receptor-2 (VEGFR2) gene expression by redundant oligonucleotide DNA microarray and real-time PCR technologies. Circulating angiogenic progenitor cells expressing VEGFR2 were profiled by flow cytometry. Plasma and serum concentrations of VEGF, nitrates/nitrites, catecholamines, and dopamine were measured by enzyme-linked immunosorbent assay (ELISA) and high performance liquid chromatography (HPLC). Arterial blood pressure, cardiac output, oxygen consumption and total peripheral resistance were determined at Baseline, 100W, and peak ergometric stress by standard techniques. There were no apparent differences (p < .05) observed in biochemical markers relating to vascular function and adaptation including, serum nitrates/nitrites, norepinephrine, dopamine, and plasma VEGF. No differences were found relative to cardiac output, stroke volume, cardiopulmonary or myocardial oxygen consumption, expired ventilation, heart rate, arteriovenous oxygen difference, total peripheral resistance, and mean arterial pressure. Due to methodological issues related to the redundant oligonucleotide DNA microarray and real-time PCR gene expression analyses, results of these experiments were uninterpretable. Thus, the research hypothesis was rejected. Conversely, significant (p < .05) differences were observed in waist: hip ratios, recovery: peak systolic blood pressure ratio at 1 minute post-exercise and %VEGFR2 expression. OSA was associated with elevations in both waist: hip ratios and recovery: peak systolic blood pressure ratio at 1 minute post-exercise as well as significant depression of %VEGFR2 profiles. Moreover, significant negative correlations were found regarding waist: hip ratios and %VEGFR2 expression (r = -.69;p =.005) and recovery: peak systolic blood pressure ratio at 1 minute post-exercise and %VEGFR2 expression (r = -.65;p =.01). These findings did not provide evidence that NO-dependent vasoactive mechanisms are suppressed nor did they support the supposition that angiogenic mechanisms are pathologically activated in sleep-disordered breathing. / Ph. D.

This dissertation comprises multiple studies surrounding the middle atmosphere's chemistry, composition, and dynamics. The middle atmosphere refers to the region from ~ 10 km to ~ 100 km and consists of the Stratosphere, Mesosphere, and Lower Thermosphere. The Stratosphere, Mesosphere, and Thermosphere are bounded by pauses where the strongest changes in chemical composition, movement, density, and thermal behavior take place. While several studies in the past have investigated the chemical composition of the middle atmosphere and quantified the distribution of various species from the stratosphere to the lower thermosphere, seasonal variations and redistribution of species resulting from transport events make it important to continuously monitor the middle atmosphere. Dynamic events such as Sudden Stratospheric Warmings (SSW) impact the temperature gradient and the zonal mean wind pattern in the stratopause. Descent events triggered by SSWs result in enhanced transport of species from the lower thermosphere to the stratosphere. Temperature increments during SSWs have an important impact on Polar Stratospheric Clouds (PSCs), resulting in Antarctic ozone enhancement and a smaller ozone hole. The middle atmosphere is, thus, home to a diverse range of dynamics and chemistry, making it a critical subject that warrants attention from the science community. The continuous monitoring of the middle atmosphere is important to this end. Several satellite missions in the past have been dedicated to monitoring the middle atmosphere and collecting data for decades. However, continual revisions and revaluations of measurement approaches and the introduction of novel space instruments are necessary to compensate for the limitations associated with existing missions, expand the extant specimen database, and improve phenomenon-centric observations. The Solar Occultation for Ice Experiment (SOFIE) is one of the two instruments on the Aeronomy of Ice in the Mesosphere (AIM) spacecraft. The studies presented in this dissertation primarily focus on the use of SOFIE observations combined with results from other science missions, an atmospheric model, and other datasets. Chapter I is an overview of the research goals and the motivations that propelled this research. In Chapter II, a validation study of the Version 1.3 SOFIE ozone data against the Atmospheric Chemistry Experiment (ACE) and the Michelson Interferometer for Passive Atmospheric Sounding (MIPAS) ozone data is presented. The SOFIE-ACE and SOFIE-MIPAS data pairs demonstrate similar variability in the ozone vertical profile. SOFIE vertical ozone profiles agree best with ACE from 30 - 70 km and MIPAS from 30-64 km. The mean difference values averaged over all seasons and both hemispheres are typically < 24% with ACE and < 20 % with MIPAS. Atomic oxygen is an important species in the mesopause region (~ 80 – 100 km) that impacts the region's ozone photochemistry and radiative balance. In Chapter III, SOFIE ozone measurements used to derive daytime atomic oxygen are compared to coincident retrievals from the Sounding of the Atmosphere using Broadband Emission Radiometry (SABER) instrument and the Naval Research Laboratory Mass Spectrometer Incoherent Scatter radar (NRLMSIS 2.0) model. The datasets agree qualitatively. Results indicate a strong seasonal variation of atomic oxygen with summer and wintertime maxima at ~ 84 km and 94 km, respectively. The middle atmospheric composition is redistributed by the transport of species during SSWs. In Chapter IV, the 2019 SSW in the northern hemisphere that triggered a large transport event from the lower thermosphere to the stratosphere is evaluated using SOFIE, ACE, and the Modern-Era Retrospective analysis for Research and Applications (MERRA-2) observations. The event was similar to the major SSW-triggered descent events in the northern hemisphere since 2004 and led to the enhancement of nitric oxide produced by Energetic Particle Precipitation, attributed to unusual meteorology. The transport peak descended by ~ 5-6 km every 10 days. An SSW event occurred in the southern hemisphere in 2019 and led to enhanced ozone in the stratosphere. In Chapter V, satellite instruments, ground station data, and measurements from NASA Ozone Watch are used to conclude that large temperature increments evaporated PSCs, resulting in the lower conversion of halogen reservoir species into ozone-destroying forms. Thus, a large ozone enhancement was recorded in 2019. Chapter VI concludes all findings and Chapter VII summarizes future work. / Doctor of Philosophy / The middle atmosphere is the region between ~ 10 and 100 km in the atmosphere and is comprised of the Stratosphere, Mesosphere, and Lower Thermosphere. The middle atmosphere is a dynamic region, and the chemistry of this region is subject to variations occurring naturally or those triggered by anomalous events such as Sudden Stratospheric Warmings (SSW). Several species in the middle atmosphere need to be measured continuously or reevaluated for improved understanding. Dynamical events in the middle atmosphere are responsible for transporting and redistributing species in the middle atmosphere. Thus, the continuous monitoring of the middle atmosphere is necessary. Novel approaches with improved techniques and approaches are thus important to explore the middle atmosphere and quantify the chemistry of the region. The Solar Occultation for Ice Experiment (SOFIE) instrument is an instrument onboard the Aeronomy of Ice in the Mesosphere (AIM) spacecraft. SOFIE typically measures at high latitudes and looks at a wide range of wavelengths. This dissertation uses SOFIE and other datasets to evaluate the varying chemistry and dynamics of the middle atmosphere. The dissertation addresses four research problems and assimilates them to evaluate the middle atmosphere. Ozone is an important species in the middle atmosphere, which is present in the highest quantity in the stratosphere, followed by the lower thermosphere (~ 85 – 100 km). Ozone is important as it absorbs ultraviolet radiations and impacts the stratospheric radiative balance. Missions in the past have monitored ozone in the middle atmosphere. Novel approaches and improved observation techniques to compensate for the limitations of past missions and the continuous measurement of ozone are necessary. Thus, ozone retrievals from SOFIE are validated against independent and established datasets to demonstrate the robustness and usability of the SOFIE ozone data product within the atmospheric science community. Atomic oxygen is an important species in the mesopause region (~ 80 – 100 km) because of its role in ozone photochemistry and impact on the radiative balance of the region. It is technologically challenging to make direct measurements of atomic oxygen; thus, most conventionally, derived measurements and model results are used. To this date, atomic oxygen has been understood in a limited capacity with several inaccuracies. To improve the understanding of atomic oxygen and fill the current knowledge gaps, atomic oxygen is derived from SOFIE ozone measurements during the daytime using the Chapman equations for ozone photochemistry. Further, the derived atomic oxygen is compared to other established datasets from satellite instrument-derived measurements and model predictions. The seasonal variability of atomic oxygen is evaluated with a focus on the difference in its behavior during summer and winter. Lastly, inter-hemispheric differences in atomic oxygen distribution are evaluated. Apart from the natural atmospheric variation in species, SSW-triggered transport events redistribute species in the atmosphere. The 2019 SSW event in the northern hemisphere was similar to those in 2004, 2006, 2009, and 2013. Large quantities of nitric oxide were transported from the lower thermosphere to the stratosphere. Air poor in water vapor and methane was also transported. Atomic oxygen was transported from the lower thermosphere to several kilometers below in amounts higher than usual. The increased nitric oxide concentration in the stratosphere due to the transport catalytically destroyed the ozone in the region. The vertical transport rates were calculated to understand the speed of the descent. The low geomagnetic index in 2019, like in all years besides 2004, indicates that these events are attributed to unusual meteorology. An SSW event took place in the southern hemisphere in 2019 during the Antarctic winter. This led to a large increase in temperature, which evaporated the Polar Stratospheric Clouds (PSCs). PSCs provide their surface for converting halogen reservoir species into ozone-destroying reactive forms. The absence of PSCs during and immediately after the SSW event led to a lower conversion of halogen reservoir species into reactive forms. Satellite instrument measurements agree with theoretical expectations. The 2002 SSW in the SH led to similar outcomes and are compared to the 2019 event. Large enhancements in ozone in 2019 led to the smallest ozone hole since ~ 1982.

Yes / Sublethal carbon monoxide (CO) exposure is frequently associated with myocardial arrhythmias, and our recent studies have demonstrated that these may be attributable to modulation of cardiac Na+ channels, causing an increase in the late current and an inhibition of the peak current. Using a recombinant expression system, we demonstrate that CO inhibits peak human Nav1.5 current amplitude without activation of the late Na+ current observed in native tissue. Inhibition was associated with a hyperpolarizing shift in the steady-state inactivation properties of the channels and was unaffected by modification of channel gating induced by anemone toxin (rATX-II). Systematic pharmacological assessment indicated that no recognized CO-sensitive intracellular signaling pathways appeared to mediate CO inhibition of Nav1.5. Inhibition was, however, markedly suppressed by inhibition of NO formation, but NO donors did not mimic or occlude channel inhibition by CO, indicating that NO alone did not account for the actions of CO. Exposure of cells to DTT immediately before CO exposure also dramatically reduced the magnitude of current inhibition. Similarly, L-cysteine and N-ethylmaleimide significantly attenuated the inhibition caused by CO. In the presence of DTT and the NO inhibitor Nω-nitro-L-arginine methyl ester hydrochloride, the ability of CO to inhibit Nav1.5 was almost fully prevented. Our data indicate that inhibition of peak Na+ current (which can lead to Brugada syndrome-like arrhythmias) occurs via a mechanism distinct from induction of the late current, requires NO formation, and is dependent on channel redox state. / This work was supported by the British Heart Foundation

Unexpectedly high concentrations of NO₂ have been noted in stack emissions from industrial gas turbines. NO₂ formation appears to occur through the so called "HO₂ mechanism II in which NO combines with HO₂ to produce NO₂ and OH. In this study, the formation of NO₂ was investigated through computer modeling and experimental testing. Computer modeling utilized the CHEMKIN chemical kinetics program and a subset of a previously published C-H-O-N system mechanism. Experimental work was conducted using a high pressure flow reactor designed and built in the course of the study. The effects of pressure, temperature, and the presence of a NO₂ promoting hydrocarbon, methane, were investigated. It was discovered that as pressure increased from 1 atm. to 8.5 atm., the rate and amount of NO converted to NO₂ also increased. There also appeared to be a temperature "window" between approximately 800 and 1000 K in which NO to NO₂ conversion readily occurred. The presence of methane was seen to enhance NO conversion to NO₂, and a ratio of [CH₄]/[NO] was found to be a useful parameter in predicting NO₂ formation. Significant NO conversion to NO₂ was noted for [CH₄]/[NO] > 1 at the hydrocarbon injection point. Experimental results validated those trends obtained from modeling with a modified C-H-O-N mechanism. / Master of Science

The effect of temporal variations in equivalence ratio on the NO_x emissions of a premixed methane-air flame was measured in a burner. The NO_x emissions are compared among steady flames with spatially uniform equivalence ratio distributions, steady flames with spatially nonuniform equivalence ratio distributions, and unsteady flames with temporal equivalence ratio fluctuations. Time-varying equivalence ratio was measured optically, time-varying temperatures were measured with thermocouples, and mean NO_x emissions were measured by probe sampling and a chemiluminescent analyzer. These measurements quantify the effect of temporal unsteadiness and spatial nonuniformity of equivalence ratio on NO_x emissions. For lean flames, both spatial nonuniformities and temporal fluctuations in equivalence ratio contribute to an increase in NO_x emissions with respect to steady uniform flames at the same mean flame temperatures. For lean flames, higher amplitude temperature fluctuations result in larger increases in NO_x with respect to steady flames. The dissertation also describes the optical technique for nonintrusive temporal measurements of equivalence ratio fluctuations and techniques for thermocouple compensation at frequencies up to 10 Hz. / Ph. D.

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No / Cortical spreading depression (CSD) is a temporary disruption of local ionic homeostasis that propagates slowly across the cerebral cortex, and may contribute to the pathophysiology of stroke and migraine. Previous studies demonstrated that nitric oxide (NO) formation promotes the repolarisation phase of CSD, and this effect may be cyclic GMP (cGMP)-mediated. Here, we have examined how phosphodiesterase (PDE) inhibition, either alone or superimposed on NO synthase (NOS) inhibition, alters CSD and the associated changes in extracellular cGMP. Microdialysis probes incorporating an electrode were implanted into the frontoparietal cortex of anaesthetised rats for quantitative recording of CSD, pharmacological manipulations, and dialysate sampling for cGMP measurements. CSD was induced by cathodal electrical stimulation in the region under study by microdialysis. Extracellular cGMP increased, but only slightly, during CSD. Perfusion of either zaprinast or sildenafil through the microdialysis probe, at concentrations that inhibited both PDE5 and PDE9 (and possibly other PDE), increased significantly extracellular cGMP. Unexpectedly, these levels remained high when NOS was subsequently inhibited with N¿-nitro- -arginine methyl ester hydrochloride ( -NAME, 1 mM). The most interesting pharmacological effect on CSD was obtained with sildenafil. This drug altered neither CSD nor the subsequent characteristic effect of NOS inhibition, i.e. a marked widening of CSD. The fact that NOS inhibition still widened CSD in the presence of the high extracellular levels of cGMP associated with PDE inhibition, suggests that NO may promote CSD recovery, independently of cGMP formation.

No / Background: The molecular regulation of endothelial nitric oxide synthase (eNOS) in blood platelets and the signalling events induced by platelet-derived NO are poorly defined. In particular, the ability of von Willebrand factor (VWF) to stimulate cyclic guanosine monophosphate (cGMP) formation in platelets has produced conflicting data. Objectives: To determine the mechanisms leading to eNOS activation and clarify the downstream signaling pathways activated by platelet-derived NO in response to VWF. Methods: We used three independent markers of NO signaling, [3H] l-citrulline production, cGMP accrual and immunoblotting of vasodilator¿stimulated phosphoprotein (VASP) to examine the NO signaling cascade in response to VWF. Results: VWF increased NO synthesis and bioavailability, as evidenced by increased [3H] l-citrulline production and cGMP accrual, respectively. VWF-induced eNOS activation was GPIb-IX-dependent and independent of integrin ¿IIbß3. cGMP formation in response to VWF required Ca2+ mobilization, Src family kinases, phosphatidylinositol 3-kinase and phospholipase C, but not protein kinase C. This suggests that a cross-talk between the signaling mechanisms regulates platelet activation and NO synthesis. VWF-induced cGMP accrual was completely blocked by apyrase and indomethacin, demonstrating an essential role for platelet-derived ADP and thromboxane A2 (TxA2). Elevated cGMP levels led to increased VASP phosphorylation at serine239 that was both protein kinase G (PKG)- and protein kinase A (PKA)-dependent. Conclusions: We demonstrate that VWF activates eNOS through a specific Ca2+-dependent GPIb receptor-signaling cascade that relies on the generation of platelet-derived ADP and TxA2. Furthermore, we provide the first evidence to suggest that platelet derived-NO/cGMP activates PKA in addition to PKG.

Yes / Exposure to CO causes early afterdepolarization arrhythmias. Previous studies in rats have indicated that arrhythmias arose as a result of augmentation of the late Na+ current. The purpose of the present study was to examine the basis for CO-induced arrhythmias in guinea pig myocytes in which action potentials (APs) more closely resemble those of human myocytes. Whole-cell current- and voltage-clamp recordings were made from isolated guinea pig myocytes as well as from human embryonic kidney 293 (HEK293) cells that express wild-type or a C723S mutant form of ether-a-go-go–related gene (ERG; Kv11.1). We also monitored the formation of peroxynitrite (ONOO−) in HEK293 cells fluorimetrically. CO—applied as the CO-releasing molecule, CORM-2—prolonged the APs and induced early afterdepolarizations in guinea pig myocytes. In HEK293 cells, CO inhibited wild-type, but not C723S mutant, Kv11.1 K+ currents. Inhibition was prevented by an antioxidant, mitochondrial inhibitors, or inhibition of NO formation. CO also raised ONOO− levels, an effect that was reversed by the ONOO− scavenger, FeTPPS [5,10,15,20-tetrakis-(4-sulfonatophenyl)-porphyrinato-iron(III)], which also prevented the CO inhibition of Kv11.1 currents and abolished the effects of CO on Kv11.1 tail currents and APs in guinea pig myocytes. Our data suggest that CO induces arrhythmias in guinea pig cardiac myocytes via the ONOO−-mediated inhibition of Kv11.1 K+ channels. / British Heart Foundation