In humans, von Willebrand factor (vWF) is a key component in hemostasis and acts as a 'cellular adhesive' by letting the circulating platelets bind to exposed subendothelium. It also acts as a carrier and stabilizer of factor VIII (FVIII). A dysfunction or reduction of vWF leads to von Willebrand disease (vWD), resulting in bleeding phenotype which affects 1% of the population. Currently there are a variety of animal models used for the study of vWF and vWD; however, they do not possess the advantages found in zebrafish. Therefore, we set out to establish zebrafish as a model for the investigation of vWF and vWD through the use of bioinformatics and various molecular techniques. Using bioinformatics we found that the vWF gene is located on chromosome 18, that the GPIb? protein sequence is conserved. Confirmation of vWF production was shown by means of immunostaining and by RT-PCR, in thrombocytes as well as in veins and arteries. Evidence of vWF involvement in hemostasis and thrombosis was shown using MO and VMO technology to produce a vWD like phenotype, resulting in an increase in TTO and TTA, as well as a reduction in FVIII when blood was tested using the kPTT assay, coinciding …

Clustering techniques are important for gene expression data analysis. However, efficient computational algorithms for clustering time-series data are still lacking. This work documents two improvements on an existing profile-based greedy algorithm for short time-series data; the first one is implementation of a scaling method on the pre-processing of the raw data to handle some extreme cases; the second improvement is modifying the strategy to generate better clusters. Simulation data and real microarray data were used to evaluate these improvements; this approach could efficiently generate more accurate clusters. A new feature-based algorithm was also developed in which steady state value; overshoot, rise time, settling time and peak time are generated by the 2nd order control system for the clustering purpose. This feature-based approach is much faster and more accurate than the existing profile-based algorithm for long time-series data.

Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most abundant environmental pharmaceutical contaminants. In this study, a proteomic analysis was conducted to identify proteins differentially expressed in gill tissue of zebrafish (Danio rerio) after a 14-day exposure to the NSAIDs ibuprofen or naproxen. A total of 104 proteins with altered expression as indicated by 2-dimensional electrophoresis were analyzed by liquid chromatography with ion trap mass spectrometry (MS/MS). A total of 14 proteins fulfilled our requirements for identification which included consistency among replicate gels as well as successful MS/MS ion searches with the MASCOT database. The most prominent feature of the differential protein expression observed after NSAID exposure was an up-regulation of proteins belonging to the globin family which are involved in the transport of oxygen from gills and availability of heme molecules required for synthesis of cyclooxygenase. Differential expression was observed at exposure concentrations as low as 1-10 µg/L indicating that altered gene expression may occur in fish subjected to environmentally realistic levels of NSAID exposure.

Adult chickens employ endothermy – internal generation of heat that maintains a constant body temperature (Tb). Prior to hatching, chicken embryos are ectothermic - controlling Tb by external heat sources. Upon hatching, the hatchling transitions from an ectotherm to an endotherm that has been shown to be delayed by hypoxia. In this study, whole animal oxygen consumption () and liver, heart, and skeletal muscle citrate synthase activity (CSA) and were measured during this transition to endothermy in chickens incubated in normoxia and hypoxia (15% O2). The only significant differences in occurred in 48 hour old hatchlings where was lower in normoxic hatchlings. There were no differences in CS activity between age and incubation oxygen levels. Additionally, preliminary 2-D protein gels of embryo and hatchling liver show changes in the proteome upon hatching. Results suggest that hypoxia had no significant effect on CSA and a minimal effect on .

The pyrimidine biosynthetic pathway is essential and similar in all bacteria. The pathway from Pseudomonas is regulated by nucleotides which bind to the upstream region of the pyrBC’ gene complex. Work in our lab mapped the genes and showed that the pyrB and pyrC’ were part of an overlap complex. The Pseudomonas aeruginosa has one circular chromosome. A former Pseudomonas now called Burkholderia cepacia is similar to P. aeruginosa except that it contains three circular chromosomes (CI, CII, CIII) and one large plasmid. The primary chromosome named CI contains the pyrBC’. To our knowledge there has been no report of the activity of ATCase in Pseudomonas and contrasted with that of Burkholderia. Here, we compare the activity of ATCase in P. aeruginosa and B .cepacia. Cells of both organisms were grown in Pseudomonas minimal medium and in Enriched medium. The ATCase was extracted and partially purified from each sample. It is hypothesized that the B. cepacia has greater activity for ATCase than do the Pseudomonas.

Research continues to show that pharmaceutical environmental contamination has caused adverse environmental effects, with one of the most studied effects being feminization of fish exposed to pharmaceutical endocrine disruptors. Additionally, there are also public health risks associated with pharmaceuticals because in-home reserves of medications provide opportunities for accidental poisoning and intentional medication abuse. Pharmaceutical take back programs have been seen as a remedy to these concerns; however a thorough review of peer-reviewed literature and publicly available information on these programs indicates limited research has been conducted to validate these programs as a purported solution. Furthermore, there are significant data gaps on key factors relating to take back program participants. The purpose of this dissertation was therefore to address these gaps in knowledge and ultimately determine if take back programs could actually improve public and environmental health. This was accomplished by conducting social and scientific research on a take back program called Denton Drug Disposal Day (D4). Socioeconomic, demographic, and geographic characteristics of D4 participants were investigated using surveys and geographic analysis. Impacts on public health were determined by comparing medications collected at D4 events with medications reported to the North Texas Poison Center as causing adverse drug exposures in Denton …

Pyrmnesium parvum, commonly known as golden algae, is an algal species that under certain circumstances releases toxins which can lead to fish kills and the death of other economically and ecologically important organisms. One of the major objectives of the study was to investigate whether a relationship exists between land use and Prymnesium parvum abundance in littoral sites of Lake Texoma, USA. Another objective was to investigate whether a relationship exists between other physical variables and counts of P. parvum. Lastly, developing a valid model that predicts P. parvum abundance was an objective of the study. Through stepwise regression, a small but highly significant amount of the variation in P. parvum counts was found to be explained by wetlands, soil erodibility and lake elevation. The developed model provides insight for potential golden algae management plans, such as maintaining wetlands and teaching land owners the relationship between soil erosivity and golden algae blooms.

This thesis utilizes a theoretical framework which links biophysical and social domains of ecosystems via ecosystem services (ES), in order to conduct a socio-ecological assessment of urban watersheds in three communities in Chilean and Argentine regions of southern Patagonia. Results from this study show that expanding urban areas may be undermining the ability of local watersheds to provide for high quality ES posing potential risks to community wellbeing. Secondly, researchers and decision makers influencing regional natural resource management share similar values to general community members but do not capture the diversity of values that exist within the broader community, and dialogue between these groups on management issues is poor. A community-based management structure is recommended for the creation of adaptive and locally relevant management strategies.