Factor VII, the initiator of the extrinsic coagulation cascade, circulates in human plasma mainly in its zymogen form, Factor VII and in small amounts in its activated form, Factor VIIa. However, the mechanism of initial generation of Factor VIIa is not known despite intensive research using currently available model systems. Earlier findings suggested serine proteases Factor VII activating protease, and hepsin play a role in activating Factor VII, however, it has remained controversial. In this work I estimated the levels of Factor VIIa and Factor VII for the first time in adult zebrafish plasma and also reevaluated the role of the above two serine proteases in activating Factor VII in vivo using zebrafish as a model system. Knockdown of factor VII activating protease did not reduce Factor VIIa levels while hepsin knockdown reduced Factor VIIa levels. After identifying role of hepsin in Factor VII activation in zebrafish, I wanted to identify novel serine proteases playing a role in Factor VII activation. However, a large scale knockdown of all serine proteases in zebrafish genome using available knockdown techniques is prohibitively expensive. Hence, I developed an inexpensive gene knockdown method which was validated with IIb gene knockdown, and knockdown all serine proteases …

The Trinity River is an urbanized, effluent-dominated river, and is heavily relied upon for drinking water. The benthic macroinvertebrate community has been monitored for over 20 years, with the focus of this dissertation on three studies (1987-88, 2005, and 2011). Water quality improvement following dechlorination resulted in increased benthic metrics. Overall habitat quality, in-stream cover, surface water total organic carbon, sediment total organic carbon, near-field urban land-use, near-field forested land-use, surface water surfactant toxic units, and depositional sediment biofilm fatty acids all have statistically significant relationships with benthic macroinvertebrate metrics. These relationships are better defined with increased taxonomic resolution at the genus/species level for all benthic taxa, including Chironomidae and Oligochaeta. It is recommend that benthic identifications for state and city water quality assessments be done at the genus/species level. A novel method for quantifying depositional sediment biofilm fatty acids has been produced and tested in this dissertation. Benthic metrics are directly related to fatty acid profiles, with several essential fatty acids found only at upstream sites.

Symbiotic nitrogen fixation occurs in plants harboring nitrogen-fixing bacteria within the plant tissue. The most widely studied association is between the legumes and rhizobia. In this relationship the plant (legumes) provides the bacteria (rhizobia) with reduced carbon derived from photosynthesis in exchange for reduced atmospheric nitrogen. This allows the plant to survive in soil, which is low in available of nitrogen. Rhizobia infect and enter plant root and reside in organs known as nodules. In the nodules the bacteria fix atmospheric nitrogen. The association between the legume, Medicago truncatula and the bacteria Sinorhizobium meliloti, has been studied in detail. Medicago mutants that have defects in nodulation help us understand the process of nitrogen fixation better. One such mutant is the Mtnip-1. Mtnip-1 plants respond to S. meliloti by producing abnormal nodules in which numerous aberrant infection threads are produced, with very rare rhizobial release into host plant cells. The mutant plant Mtnip-1 has an abnormal defense-like response in root nodules as well as defects in lateral root development. Three alleles of the Mtnip/latd mutants, Mtnip-1, Mtlatd and Mtnip-3 show different degrees of severity in their phenotype. Phylogenetic analysis showed that MtNIP/LATD encodes a protein belonging to the NRT1(PTR) family of …

Natural history collections house immense amounts of data, but the majority of data is only accessible by locating the collection label, which is usually attached to the physical specimen. This method of data retrieval is time consuming and can be very damaging to fragile specimens. Digitizing the collections is the one way to reduce the time and potential damage related to finding the collection objects. The Elm Fork Natural Heritage Museum is a natural history museum located at the University of North Texas and contains collections of both vertebrate and invertebrate taxa, as well as plants. This project designed a collection digitization workflow for Elm Fork by working through digitizing the Benjamin B. Harris Herbarium. The collection was cataloged in Specify 6, a database program designed for natural history collection management. By working through one of the museum’s collections, the project was able to identify and address challenges related to digitizing the museum’s holdings in order to create robust workflows. The project also produced a series of documents explaining common processes in Specify and a data management plan.

Due to the wide spread occurrence of medroxyprogesterone acetate (MPA), a pharmaceutical compound, in wastewater effluent and surface waters, the objectives of this work were to determine the tissue specific uptake and bioconcentration factor (BCF) for MPA in common carp. BCFs were experimentally determined for MPA in fish using a 14-day laboratory test whereby carp where exposed to 100 μg/L of MPA for a 7-day period followed by a depuration phase in which fish were maintained in dechlorinated tap water for an additional 7 days. MPA concentrations in muscle, brain, liver and plasma were determined by liquid chromatography/mass spectrometry (LC/MS). The results from the experiment indicate that MPA can accumulate in fish, however, MPA is not considered to be bioaccumulative based on regulatory standards (BCF ≥ 1000). Although MPA has a low BCF value in common carp, this compound may cause reproductive effects in fish at environmentally relevant concentrations.

Glyphosate based herbicides are among the most widely used herbicides in the world. The purpose of this study was to determine developmental toxicity of glyphosate, the active ingredient in the common herbicide Roundup, on developing chicken embryos. Few studies have examined toxic effects of glyphosate alone versus the full compound formulations of Roundup, which include adjuvants and surfactants. Adjutants and surfactants are added to aid in solubility and absorption of glyphosate. In this study chicken embryos were exposed at the air cell on embryonic day 6 to 19.8 or 9.9 mg / Kg egg mass of glyphosate in Roundup or glyphosate only. Chickens treated with 19.8 and 9.9 mg / Kg glyphosate in Roundup showed significant reduction in survivability compared to glyphosate alone treatments and controls. On embryonic day 18, embryos were sacrificed for evaluation of developmental toxicity using wet embryo mass, dry embryo mass, and yolk mass as indicators. Morphology measurements were taken on liver mass, heart mass, tibiotarsus length and beak length. Embryos treated with 19.8 mg / Kg glyphosate and 9.9 mg / Kg glyphosate in Roundup showed significant reductions in wet and dry embryo mass and yolk mass. Tibiotarsus length in 9.9 mg / Kg glyphosate …

Arbuscular mycorrhizal fungi, colonizing over 80% of all plants, were long thought absent in wetlands; however, recent studies have shown many wetland plants harbor arbuscular mycorrhizae (AM) and dark septate endophytes (DSE). Wetland services such as biodiversity, shoreline stabilization, water purification, flood control, etc. have been estimated to have a global value of $14.9 trillion. Recognition of these vital services is accompanied by growing concern for their vulnerability and continued loss, which has resulted in an increased need to understand wetland plant communities and mycorrhizal symbiosis. Factors regulating AM and DSE colonization need to be better understood to predict plant community response and ultimately wetland functioning when confronting natural and human induced stressors. This study focused on the effects of water quality, hydrology, sedimentation, and hurricanes on AM and DSE colonization in three wetland species (Taxodium distichum, Panicum hemitomon, and Typhal domingensis) and plant communities of coastal wetlands in Southeast Louisiana and effects of an antimicrobial biocide, triclosan (TCS), on AM (Glomus intraradices) spore germination, hyphal growth, hyphal branching, and colonization in fresh water wetland plants (Eclipta prostrata, Hibiscus laevis, and Sesbania herbacea) from bottom land hardwood forest in north central Texas. The former, mesocosm studies simulating coastal marsh vegetation …

Improvements in mass spectrometry (MS)-based strategies for characterizing the plant lipidome through quantitative and qualitative approaches such as shotgun lipidomics have substantially enhanced our understanding of the structural diversity and functional complexity of plant lipids. However, most of these approaches require chemical extractions that result in the loss of the original spatial context and cellular compartmentation for these compounds. To address this current limitation, several technologies were developed to visualize lipids in situ with detailed chemical information. A subcellular visualization approach, direct organelle MS, was developed for directly sampling and analyzing the triacylglycerol contents within purified lipid droplets (LDs) at the level of a single LD. Sampling of single LDs demonstrated seed lipid droplet-to-droplet variability in triacylglycerol (TAG) composition suggesting that there may be substantial variation in the intracellular packaging process for neutral lipids in plant tissues. A cellular and tissue visualization approach, MS imaging, was implemented and enhanced for visualizing the lipid distributions in oilseeds. In mature cotton seed embryos distributions of storage lipids (TAGs) and their phosphatidylcholine (PCs) precursors were distribution heterogeneous between the cotyledons and embryonic axis raising new questions about extent and regulation of oilseed heterogeneity. Extension of this methodology provides an avenue for understanding metabolism …

A study of macroinvertebrate colonization and assemblages, including secondary productivity of the familiar bluet damselfly or Enallagma civile Hagen (Odonata: Coenagrionidae), associated with the aquatic macrophytes Heteranthera dubia (Jacq.) MacMill. (water stargrass) and Potamogeton nodosus Poir. (American pondweed) was conducted at the Dallas Floodway Extension Trinity River Project (DFE) Lower Chain of Wetlands (LCOW), Dallas, TX, from September 2010 through November 2011. Macroinvertebrate abundance, taxa richness, Simpson's index of diversity, and Simpson's evenness from the two macrophytes and from three different wetland cells of varying construction completion dates, water sources, and native aquatic vegetation establishment were analyzed along with basic water quality metrics (temperature °C, pH, dissolved oxygen mg/L, and conductivity µs/cm). E. civile nymphs were separated into five developmental classes for secondary productivity estimations between macrophytes and wetland cell types. Mean annual secondary productivity in the DFE LCOW among two macrophytes of E. civile was 1392.90 ash-free dry weight mg/m²/yr, standing stock biomass was 136.77 AFDW mg/m2/yr, cohort production / biomass (P/B) ratio was calculated to be 4.30 / yr and the annual production / biomass (P/B) ratio was 10.18 /yr.