Although tachykinins have potent vascular actions, their effect on renal resistance blood vessels is currently unknown. The vasoactive properties of tachykinins and related analogs were assessed in isolated perfused rat kidney. At a basal perfusion pressure (PP) of 75 $\pm$ 6 mm Hg (n = 5), bolus injections of substance P (SP) had no significant vasoactive effect. Following a sustained increase in baseline PP (134 $\pm$ 10 mm Hg) produced by phenylephrine (1 $\mu$M), SP evoked a dose-dependent increase in PP. The largest dose of SP increased PP by 60 $\pm$ 5 mm Hg. The vasoconstrictor response to SP was not blocked by phentolamine when angiotensin II was used to increase basal tone. Thus, the response to SP is not mediated by norepinephrine. Pressor responses to SP were not potentiated by peptidase inhibitors, captopril and thiorphan. SP(1-7) had no effect on PP, suggesting that the pressor response to SP is C-terminal dependent and tachykinin receptor mediated. The selective NK-1 receptor agonist, (Sar$\sp9$,Met(O$\sb2)\sp{11}\rbrack$SP, had no effect on PP. In contrast, both the selective NK-2 and NK-3 receptor agonists, GR-64349 and (MePhe$\sp7$) NKB, produced dose-dependent pressor responses (116 $\pm$ 8 and 134 $\pm$ 15 mm Hg increases in PP at 33 nmol, respectively) and were more potent than SP. Infusion of capsaicin (500 nM) produced an initial increase in PP following by a more prolonged decrease in PP. Clamping the renal vein produced a marked increase in PP. The localization of NK-3 receptors in rat kidney evaluated by film autoradiography using $\sp{125}$I- (MePhe$\sp7\rbrack$NKB revealed a high density of specific binding sites on the proximal ureter and renal pelvis, moderate density in the renal vein and its large branches, and a low density in the inner strip of outer medulla, but no specific binding on the renal artery system and cortex. High resolution autoradiograms demonstrated $\sp{125}$I- (MePhe$\sp7\rbrack$NKB binding sites on the tunica media of the renal vein and tunica muscularises of renal pelvis and ureter. Specific binding of $\sp{125}$I-BHSP was found in association with the renal artery and renal pelvis. No specific SP binding sites were associated with renal vein. These data indicate that the pressor effect of tachykinins in the isolated rat kidney can be mediated by NK-2 and/or NK-3 receptors. The latter may be on the vascular smooth muscle of the renal vein.

Tumor necrosis factor α (TNFα) is a pleiotropic cytokine that is predominantly produced by activated macrophages. The effects of TNFα are as diverse as the cells with which it interacts, e.g., stimulating fibroblast growth, exerting cytotoxic/cytostatic; activity against various human and murine cell lines, promoting inflammation through upregulation of endothelial adhesion molecules and IL-8 production. Yet TNFα is best known, and in fact was originally described, for its role in the bacterial-induced hemorrhagic necrosis of tumors and exacerbation of septic shock in which aberrant TNFα production leads to vascular collapse, cachexia, multiple organ failure, and ultimately death in as many as 100,000 people each year in the United States alone. LPS, a component of the outer cell wall of gram-negative bacteria, is the principal inducer of macrophage TNFα production. TNFα production can be enhanced by IFNγ which also induces upregulation of TNFα receptors allowing for the establishment of a TNFα autocrine loop. It has been hypothesized that autocrine TNFα stimulation plays a critical role in the induction of macrophage effector function, e.g., nitric oxide production. This dissertation represents efforts to evaluate the respective roles of the TNFα receptors in the induction of macrophage effector function, in addition to examining the mechanism by which autocrine TNFα exerts its effects on macrophages. Exploiting the species specificity of the murine TNFα receptor type 2 (TNF-R2), splenic macrophages were stimulated with human TNFα (which binds to TNF-R1 but not TNF-R2), in the presence of IFNγ. Human TNFα was effective in the induction of nitric oxide production, albeit at concentrations 12.5-fold greater than those required by murine TNFα to achieve the same result. Addition of anti-TNF-R1 completely inhibited the murine TNFα mediated induction of macrophage effector function. However, treatment with anti-TNF-R2 resulted in partial inhibition of macrophage activation. Taken together this data suggests that the primary TNFα mediated signals involved in macrophage activation are transduced through TNF-R1, although TNF-R2 appears to contribute to the intensity of the macrophage response. To evaluate the role of autocrine TNFα signaling in the induction of macrophage effector function, immortalized macrophages from normal C57Bl/6J mice (B6/J2) and C57Bl/6J mice containing gene targeted disruptions of the TNF-R1 and TNF-R2 genes (TRN) were stimulated under CD14-dependent and CD14-independent conditions. Although the B6/J2 and TRN clones mounted similar NO responses to LPS in the presence of serum, the TRN macrophages generated a weak nitric oxide response as compared to B6/J2 when stimulated with LPS under serum-free conditions. The involvement of TNFα autocrine stimulation in the CD14-independent activation was corroborated by the ability of soluble TNF-R1 to inhibit the response of B6/J2 macrophages to LPS in serum-free medium. CD14-independent LPS stimulation of TRN and B6/J2 resulted in equivalent levels of IL-1β, TNFα, and NOS gene expression, as determined by RT-PCR, and in release of equivalent amounts of biologically active TNFα. However, western blot analysis revealed that NOS protein production by TRN was as much as 50% less than that produced by B6/J2. These results indicate that autocrine TNFα stimulation contributes to the signaling pathways initiated by ligation of CD14-independent LPS receptors and may be involved in NOS post-transcriptional regulation.

The purpose of this study was to compare Registered Nurse (RN) current and future demand between Tennessee rural and urban licensed hospitals and nursing homes. Comparisons of Registered Nurse demand by principal duty or position and by educational level were also undertaken. The educational level of RNs was examined using the proportion of RNs at each of five educational levels: diploma, associate, baccalaureate, master's, and doctorate. The variables examined were 1988 and 1989 vacancy rates and proportion of budgeted positions, change in vacancy rates and proportions from 1988 to 1989, projected changes in positions and proportions from 1988 to 1992 and to 1995. Nurse administrators from one hundred fifty four hospitals (63% response rate) and two hundred twelve nursing homes (72% response rate) were surveyed for responses about current and future RN demand. This study is a secondary analysis of data that were collected as a part of a larger research project conducted by the Tennessee Board of Regents Task Force on Nurse Supply and Demand. The findings were that rural hospitals had significantly higher vacancy rates for total nurse positions in 1988 and general duty positions in 1989. Urban hospital administrators reported higher vacancy rates for RN positions at the diploma level and projected significantly greater changes in clinical specialist and master's level positions for 1992 and 1995. Rural hospitals had a higher proportion of associate degree nurses in 1989, and urban hospitals had a higher percentage of master's prepared RNs for 1988 and 1989. Urban hospitals projected a greater change in the proportion of RNs at the doctoral level for 1995. Rural nursing homes projected higher numbers of associate degree nurses for 1992 and 1995. The major conclusion was that the nursing shortage in Tennessee hospitals was more severe than that reported on the national level and greater in rural hospitals. The shortage in Tennessee nursing homes was reported to be much less acute than the national shortage, and the critical shortages were limited to a few facilities, both rural and urban.

The signaling mechanisms for muscarinic receptor-mediated vasoconstriction in coronary resistance arteries were studied in KCl-arrested isolated rat hearts perfused at a constant flow rate. The cholinergic agonists acetylcholine and bethanechol were given by bolus injection or constant infusion. The coronary vascular resistance was monitored by measuring the changes in perfusion pressure. The selective muscarinic agonist bethanechol caused a similar vasoconstrictor response as ACh, but with less potency and efficacy. Bolus injection of bethanechol evoked a phasic vasoconstriction in a dose-dependent manner, while infusion of bethanechol evoked a tonic vasoconstriction without producing tachyphylaxis. Coronary vascular responses to bethanechol were further examined in the absence and presence of a specific inhibitor or blocker for the potential signaling components. The bethanechol-induced phasic vasoconstriction was eliminated by perfusion with a Ca2+ -free medium. The maximal vasoconstriction to bethanechol was suppressed by 31% in the presence of the Ca2+ -dependent Cl- -channel blocker A-9-C. The L-type voltage-operated Ca2+ channel blocker nifedipine decreased the maximal constrictor response to bethanechol by 59%, while the putative receptor-operated Ca 2+ channel blocker SK&F 96365 converted this vasoconstriction into vasodilation which was not affected by the nitric oxide synthase inhibitor L-NAME. Coronary vascular responses to bethanechol were almost abolished by a combination of nifedipine and SK&F 96365. The protein kinase C inhibitor chelerythruine reduced bethanechol-evoked peak vasoconstriction by 79%. The bethanechol-induced tonic vasoconstriction was rapidly converted into vasodilation by the concomitant infusion of SK&F 96365 or nifedipine, but the simultaneous infusion of chelerythrine gradually attenuated this response. These data suggest that the novel receptor-operated Ca2+ channel, voltage-operated Ca2+ channel, and protein kinase C are the most crucial signaling components for muscarinic receptor-mediated coronary vasoconstriction in the isolated rat heart. Extracellular Ca 2+ influx via receptor-operated Ca2+ channels and voltage-operated Ca2+ channels is essential to both phasic and tonic vasoconstrictor responses to bethanechol. Protein kinase C plays a pivotal role in the regulation of bethanechol-evoked vasoconstriction by sensitizing the contractile apparatus and modulating the activity of Ca 2+ channels.

Recent research has indicated that dissolved organic matter (DOM) may plan an important role in the ability of natural waters to complex metals. This research was conducted because the quantitative nature of this role is uncertain. Gas-liquid chromatography was used to study the hydrolyzable amino acids at twelve sampling sites on the Williamson River at monthly intervals for two years. The relative abundances showed little spacial or temporal variation. The two-year averages for total amino acids ranged from about 0.5 (mu)M to about 8 (mu)M. A separation technique was used to show that (GREATERTHEQ) 96% of the dissolved amino acids were associated with aquatic humus. Since it was found that amino acids contributed less than 1% to humic carbon and since a published report found that carbohydrates contributed less than 2% to humic carbon, this research provided the necessary data to conclude that DOM in the Williamson River is essentially aquatic humus. Humus complexation capacity is often operationally defined as amount of metal bound per unit weight of humus. This research has shown that the titrimetric methods commonly used to obtain this parameter underestimate its magnitude. However, it was shown that these methods can be combined with acidic functional group analyses to determine upper and lower limit for this parameter. For Williamson River humus, the range was 7.2 - 15.4 (mu)mols copper per mg humic carbon. Titrations of humus into a copper-oxalate metal-ion buffer enabled the determination of the copper-humus binding "constant" at humas : copper ratios found in the Williamson River, (LESSTHEQ) 4300. The binding "constant" was a variable and a function of pH. At a humus: copper ratio of 4300, the values of the function at pH 5.0, 5.5, 6.0, and 6.5 were: 3.0 x 10('6), 8.9 x 10('6), 3.0 x 10('7), and 1.7 x 10('8). Current models of metal-humus complexation, were shown to be inappropriate via rigorous mathematical examination and via application to computer-simulated titrations. A model, in which it is assumed that the concentrations of binding sites in humus are normally distributed with respect to the log of the metal binding constant for each site, is proposed. Application of this model to simulated titrations and to experimental data proved it to be superior to other current models.

Chronic exposure to some oxysterols might contribute to deterioration of human or environmental health. Oxysterols are both biomarkers of oxidative stress as well as mediators of its damage, and play a central role in many independent, but converging, disease processes, such as atherosclerosis, Alzheimer's disease, and age-related macular degeneration. Therefore, the aim of this thesis was to identify enzymes capable of transforming oxysterols to either reduce their toxicity or facilitate their metabolism or excretion. 7-ketocholesterol (7KC), being amongst the most cytotoxic and recalcitrant of these compounds, was the main focus of this work. We isolated various bacteria capable of utilizing 7KC as a sole carbon and energy source. One of these, Rhodococcus jostii RHAl , was subjected to rigorous transcriptomic and mutational analysis to elucidate its 7KC degradation pathway, which was similar, but not identical, to that of cholesterol. Metabolite screening revealed the reduction and subsequent removal of the 7-keto moiety prior to the step catalyzed by HsaC, the enzyme responsible for cleavage of sterol ring A. Furthermore, cloning and expression of a number of reductases from two gene clusters that were highly up-regulated during growth on 7KC identified three reductases that are active against several closely related structural analogs, though not 7KC itself. 7KC and a number of analogs were assayed for toxicity against human fibroblasts Several enzymes were overexpressed in these fibroblasts by transient transfection with mammalian expression vectors to screen for their ability to mitigate 7KC-induced cytotoxicity. A LAMP1/cholesterol oxidase chimera was found to be significantly cytoprotective to exposure to up to 50 μM 7KC compared to mock transfection as well as 7KC-transforming enzymes targeted to either the mitochondria or cytosol. Additionally, transfection with LAMP1 alone and treatment with 0.9% hydroxypropyl β-cyclodextrin also reduced toxicity. Therefore, it seems likely that addressing 7KC toxicity within the lysosome is critical for cytoprotection. This work provides preliminary evidence to support this approach, and may have implications for the treatment of oxysterol-associated diseases. However, further research is needed to evaluate the effects and safety of heterologous gene expression within the lysosome, both in vitro and in vivo.

Pediatric female and young adult female patients who undergo computed tomography (CT) scanning may be at higher risk for developing radiation-induced breast cancer later in life. Thus, the purpose of this thesis was to both accurately quantify dose and explore new strategies for CT breast dose reduction. In order to determine dose reduction, dose quantification was first assessed through the development and validation of an empirical model for describing attenuation in CT and second through evaluation of the precision of dosimetry-related measurements obtained using three different models of CT scanners. Breast dose-savings was evaluated using CT dose index phantoms, anthropomorphic phantoms, and Monte Carlo computer modeling. Modifications to current scanning procedures, such as proper patient centering and beginning data acquisition with the x-ray tube facing a patient's posterior, were shown to minimize breast dose. Novel techniques, including varying the x-ray tube voltage during scanning and incorporation of a dynamic x-ray beam filter over the breasts, were also found to successfully reduce breast dose.

Sustainability thinking has advanced considerably in the last few decades. Despite these advances, there remains a significant disconnect between thoughts and actions in this field. Sustainability metrics are one method proposed to help address this problem. A literature review of the most commonly used metrics and evaluation systems will first be presented, along with a critical evaluation of each system against the basic principles of sustainability. Next, a new evaluation framework will be proposed that addresses some of the shortcomings of the existing evaluation methods. This framework is designed to follow the best available thinking on a number of sustainability issues and overall, give a clear and concise evaluation of sustainability while still maintaining accuracy and scientific relevancy. Finally, a theoretical application of the framework to evaluating the sustainability of the city of Houston, Texas will be given, showing the necessary metrics required to follow the principles of the framework.

Positron Emission Tomography (PET) is a widely used imaging modality for diagnosing patients with cancer. Recently, there have been three novel developments in PET imaging aiming to increase PET image quality and quantification. This thesis focuses on the optimization of PET image quality on these three developments. The first development is the fully 3D PET data acquisition and reconstruction. 3D Acquisitions are not constrained in collecting events in single 2D planes and can span across different planes. 3D acquisition provides better detection since it can accept more events. Also it can result in lower radiation dose to the patient and shorter imaging times. With the application of 3D acquisition, a fully 3D iterative reconstruction algorithm was also developed. The aim of the first project in this thesis is to evaluate the PET image and raw data quality when this fully 3D iterative reconstruction algorithm is applied. The second development in PET imaging is the time-of-flight (TOF) PET data acquisition and reconstruction. TOF imaging has the ability to measure the difference between the detection times, thus localize the event location more accurately to increase the image quality. The second project in this thesis focuses on optimizing the TOF reconstruction parameters on a newly developed TOF PET scanner. Then the improvement of TOF information on image quality is assessed using the derived optimal parameters. Finally the effect of scan duration is evaluated to determine whether similar image quality could be obtained between TOF and non-TOF while using less scan time for TOF. The third development is the interest in building PET / magnetic resonance (MR) multi-modality scanner. MR imaging has the ability to show high soft tissue contrast and can assess physiological processes, which cannot be achieved on PET images. One problem in developing PET/MR system is that it is not possible with current MR acquisition schemes to translate the MR image into an attenuation map to correct for PET attenuations. The third project in this thesis proposed and assessed an approach for the attenuation correction of PET data in potential PET/MR systems to improve PET image quality and quantification.

This research addresses the potential ecotoxicity of two emerging carbonaceous materials: C 60 and biochar. The use of these materials is rapidly increasing, as well as their potential for widespread applications. Thus, information about unintended consequences associated the widespread use, incidental or accidental release, and disposal of these emerging materials is needed. The environmental impacts of C 60 , its stable water suspension (nC 60 ), and biochar are assessed here using bacteria and earthworms as model receptors. The antibacterial activity of nC 60 can be mitigated by the presence of natural organic matter as a soil constituent or dissolved in the water column. Sorption to soil might decrease the bioavailability of nC 60 and thus its toxicity to bacteria. Aqueous organic matter also may mitigate nC 60 toxicity. Pristine C 60 showed toxicity to the earthworm's reproduction and was rapidly bioaccumulated by earthworms, although to a lower extent than smaller phenanthrene molecules that are more hydrophobic; thus, the large molecular size of C 60 hinders its bioaccumulation. Less bioaccumulation occurred at higher C 60 concentration in soil, which is counterintuitive and reflects that higher C 60 concentrations that exceed the soil sorption capacity exist as larger precipitates that are less bioavailable. Earthworms avoided soils amended with high concentrations of dry biochar, and experienced significant weight loss after 28-day exposure. The avoidance response was likely to avert desiccation rather than to avoid potential toxicants (i.e., PAHs formed during biochar production by pyrolysis) or nutrient scarcity. By wetting the biochar to field capacity before exposing the worms, this adverse effect can be completely mitigated. Overall, this research provides a foundation for ecotoxicity assessment associated with exposure to C 60 or biochar, and establishes a method by which other emerging materials can be evaluated for their potential environmental impacts.