A study of Asian clam (Corbicula fluminea Müller) colonization in relation to changes in aquatic vegetation community as a result of management of Hydrilla verticillata (L. f.) Royle with grass carp was conducted at the Lewisville Aquatic Ecosystem Research Facility (LAERF), Lewisville, TX, from April 2015 through October 2016. Percent vegetation cover, C. fluminea abundance and water quality metrics (pH, turbidity, conductivity, DO, calcium, chlorophyll a) from 16 experimental subjects were analyzed. Treatments included four replicated grass carp stocking densities; 1-control with no fish stocked (n = 4), 2-low density of 40-43 fish per vegetated ha (n = 4), 3-medium density of 72-81 fish per vegetated ha (n = 4) and 4-high density of 110-129 fish per vegetated ha (n = 4). Data analysis showed statistical significance in the relation of C. fluminea abundance to percent vegetation cover (multiple linear regression, r2 = 0.820), grass carp stocking densities (two-way analysis of variance, p = <0.001) and chlorophyll a (multiple linear regression, r2 = 0.339). Findings of this research indicate the possibility that management of hydrilla had enabled establishment of secondary invasive species.

This thesis addresses three interrelated challenges of disease mapping and contributes a new approach for improving visualization of disease burdens to enhance disease surveillance systems. First, it determines an appropriate threshold choice (smoothing parameter) for the adaptive kernel density estimation (KDE) in disease mapping. The results show that the appropriate threshold value depends on the characteristics of data, and bandwidth selector algorithms can be used to guide such decisions about mapping parameters. Similar approaches are recommended for map-makers who are faced with decisions about choosing threshold values for their own data. This can facilitate threshold selection. Second, the study evaluates the relative performance of the adaptive KDE and spatial empirical Bayes for disease mapping. The results reveal that while the estimated rates at the state level computed from both methods are identical, those at the zip code level are slightly different. These findings indicate that using either the adaptive KDE or spatial empirical Bayes method to map disease in urban areas may provide identical rate estimates, but caution is necessary when mapping diseases in non-urban (sparsely populated) areas. This study contributes insights on the relative performance in terms of accuracy of visual representation and associated limitations. Lastly, the study contributes …

Plant architecture is an important agronomic trait driven by meristematic activities. Indeterminate meristems set repeating phytomers while determinate meristems produce terminal structures. The centroradialis/terminal flower1/self pruning (CETS) gene family modulates architecture by controlling determinate and indeterminate growth. Cotton (G. hirsutum) is naturally a photoperiodic perennial cultivated as a day-neutral annual. Management of this fiber crop is complicated by continued vegetative growth and asynchronous fruit set. Here, cotton CETS genes are phylogenetically and functionally characterized. We identified eight CETS genes in diploid cotton (G. raimondii and G. arboreum) and sixteen in tetraploid G. hirsutum that grouped within the three generally accepted CETS clades: flowering locus T (FT)-like, terminal flower1/self pruning (TFL1/SP)-like, and mother of FT and TFL1 (MFT)-like. Over-expression of single flower truss (GhSFT), the ortholog to Arabidopsis FT, accelerates the onset of flowering in Arabidopsis Col-0. In mutant rescue analysis, this gene driven by its native promoter rescues the ft-10 late flowering phenotype. GhSFT upstream sequence was used to drive expression of the uidA reporter gene. As anticipated, GUS accumulated in the vasculature of Arabidopsis leaves. Cotton has five TFL1-like genes, all of which delay flowering when ectopically expressed in Arabidopsis; the strongest phenotypes fail to produce functional flowers. Three …

Plant microtubules play essential roles in cell processes such as cell division, cell elongation, and organelle organization. Microtubules are arranged in highly dynamic and ordered arrays, but unlike animal cells, plant cells lack centrosomes. Therefore, microtubule nucleation and organization are governed by microtubule-associated proteins, including a microtubule-severing protein, katanin. Mutant analysis and in vitro characterization has shown that the highly conserved katanin is needed for the organization of the microtubule arrays in Arabidopsis and rice as well as in a variety of animal models. Katanin is a protein complex that is part of the AAA+ family of ATPases. Katanin is composed of two subunits, katanin-p60, a catalytic subunit and katanin-p80, a regulatory subunit. Spastin is another MT-severing protein that was identified on the basis of its homology to katanin. In animal cells, spastin is also needed for microtubule organization, but its functionality has not yet been investigated in plants. To initiate an exploration of the function of katanin-p60 and spastin in Zea mays, my research goal was to generate tools for the expression and purification of maize katanin-p60 and spastin proteins in vitro. Plasmids that express katanin-p60 and spastin with N-terminal GST tags were designed and constructed via In-Fusion® cloning …

Myosin subfragment-2 (S2) is a coiled coil linker between myosin subfragment-1 and light meromyosin (LMM). This dissertation examines whether the myosin S2 coiled coil could regulate the amount of myosin S1 heads available to bind actin thin filaments by modulating the stability of its coiled coil. A stable myosin S2 coiled coil would have less active myosin S1 heads compared to a more flexible myosin S2 coiled coil, thus causing increased force production through acto-myosin interaction. The stability of the myosin S2 coiled coil was modulated by the binding of a natural myosin S2 binding protein, myosin binding protein C (MyBPC), and synthetic myosin S2 binding proteins, stabilizer and destabilizer peptide, to myosin S2. Competitive enzyme linked immunosorbent assay (cELISA) experiments revealed the cross specificity and high binding affinity of the synthetic peptides to the myosin S2 of human cardiac and rabbit skeletal origins. Gravitational force spectroscopy (GFS) was performed to test the stability of myosin S2 coiled coil in the presence of these myosin S2 binding proteins. GFS experiments demonstrated the stabilization of the myosin S2 coiled coil by the binding of MyBPC and stabilizer peptide to myosin S2, while the binding of destabilizer peptide to the same resulted …

Recently, three distinct isoforms of phospholipase D (PLD) were identified in Arabidopsis thaliana. PLD α represents the well-known form found in plants, while PLD β and γ have been only recently discovered (Pappan et al., 1997b; Qin et al., 1997). These isoforms differ in substrate selectivity and cofactors required for activity. Here, I report that PLD β and γ isoforms were active toward N-acylphosphatidylethanolamine (NAPE), but PLD α was not. The ability of PLD β and γ to hydrolyze NAPE marks a key difference from PLD α. N-acylethanolamines (NAE), the hydrolytic products of NAPE by PLD β and γ, inhibited PLD α from castor bean and cabbage. Inhibition of PLD α by NAE was dose-dependent and inversely proportional to acyl chain length and degree of unsaturation. Enzyme kinetic analysis suggested non-competitive inhibition of PLD α by NAE 14:0. In addition, a 1.2-kb tobacco (Nicotiana tabacum L.) cDNA fragment was isolated that possessed a 74% amino acid identity to Arabidopsis PLD β indicating that this isoform is expressed in tobacco cells. Collectively, these results provide evidence for NAE producing PLD activities and suggest a possible regulatory role for NAE with respect to PLD α.

No apparent repression of pyr gene expression in Pseudomonas aeruginosa is observed upon addition of exogenous pyrimidines to the growth medium. Upon introduction of the subcloned Escherichia coli pyrBI genes for aspartate transcarbamoylase (ATCase) into a P. aeruginosa pyrB mutant strain, repression was observed in response to exogenously fed pyrimidine compounds. The results proved that it is possible to bring about changes in pyrimidine nucleotide pool levels and changes in transcriptional regulation of gene expression as a result. Thus, the lack of regulatory control in P. aeruginosa pyr gene expression is not due to an inability to take up and incorporate pyrimidine compounds into metabolic pools, or to an inability of the RNA polymerase to respond to regulatory sequences in the DNA but is probably due to a lack of specific regulatory signals in the promoter of the genes themselves.

The TOL plasmids of Pseudomonas putida encode enzymes required for the oxidation of toluene and other related aromatic compounds. These genes are organized into two operons, the xylUWCMABN operon (upper), and the xylXYZLTEGFJQKIH operon (lower). Here we report the nucleotide sequence of a 7107 bp segment of the TOL pDK1 plasmid encoding the region just upstream of the "upper" operon through the genes encoding xylUWCMA. Sequence analysis, comparison of base-usage patterns, codon-usage patterns, and intergenic distances between genes help support the idea that the "upper" and "lower" operons have evolved independently in different genetic backgrounds and have only more recently been brought together in TOL and related catabolic plasmids.

The purpose of the research was to treat immunodeficient mice, implanted with colon-25 tumors, with continuous and interrupted electrochemical therapy (ECT). ECT involves the placement of two electrodes, an anode near the center of the tumor and a cathode into the tumor periphery. A constant voltage is applied across the electrodes for a given period of time. The data showed that the interrupted and continuous ECT resulted in a decrease in mean tumor growth as compared to that of the sham controls. The histology of both ECT groups showed an increase presence of large vacuoles, randomly distributed tumor cells as well as the presence of "crevicing" in the medullary tissue. The differential leukocyte counts showed a distinct neutrophilia and lymphopenia in all groups at day 20 post tumor implantation. The results from the experimental groups appeared to support the findings of previous investigators.

Aspartate transcarbamoylase (ATCase) was purified from 16 selected bacterial species including existing Pseudomonas species and former species reassigned to new genera. An enormous diversity was seen among the 16 enzymes with each class of ATCase being represented. The smallest class, class C, with a catalytically active homotrimer, at 100 kDa, was found in Bacillus and other Gram positive bacteria. In this report, the ATCases from the Gram negatives, Shewanella putrefaciens and Stenotrophomonas maltophilia were added to class C membership. The enteric bacteria typify class B ATCases at 310 kDa, with a dodecameric structure composed of two catalytic trimers coupled to three regulatory dimers. A key feature of class B ATCases is the dissociability of the holoenzyme into regulatory and catalytic subunits which were enzymatically active. In this report, the ATCase from Pseudomonas indigofera was added to class B ATCases. The largest class, at 480 kDa, class A, contains the fluorescent Pseudomonas including most members of the 16S rRNA homology group I. Two polypeptides are produced from overlapping pyrBC' genes. The former, pyrB, encodes a 34 kDa catalytic polypeptide while pyrC' encodes a 45 kDa dihydroorotase-like polypeptide. Two non active trimers are made from six 34 kDa chains which are cemented …

Limitations from reduced light and increasing water depth on Nelumbo lutea seedlings were determined in tank experiments. Survival was high in all tested light levels. Total biomass increased significantly with increasing light. Biomass allocation shifted significantly to root production between 3 and 6 weeks in the 10 and 24% levels. Survival decreased with increasing planting depth, and biomass of survivors reduced significantly between 0.5, 1.0, and 1.5 m depths. Nelumbo lutea and Myriophyllum spicatum populations were monitored for one season in a 0.7 ha pond to track changes in species dominance. Myriophyllum spicatum dominated early, and N. lutea dominated from July through October, suppressing M. spicatum at all depths. Competitive interactions between N. lutea and M. spicatum were investigated for two seasons in a container experiment situated within a pond. Where established, N. lutea dominated in the presence of M. spicatum. However, N. lutea could not be established in depths greater than 1 meter.

A toxicological study was initiated because of concerns about allegations that the Texas Water Commission that effluent from the Dallas Central Wastewater Treatment Plant, which discharges into the Trinity River, was affecting downstream water quality. Monthly, flow-weighted composite effluent samples were collected. Grab samples were also collected upstream and downstream from the effluent from April 1989 to August 1991. Toxicity tests were conducted on these samples using Ceriodaphnia dubia as the test organism. Samples were collected four times during this study in which rainfall occurred prior to sampling. In every instance, this "first flush" of the watershed during a rising hydrograph was toxic to C. dubia upstream. Analyzing toxicity by season resulted in a statistically significantly lower neonate production in the effluent than in the river samples during the months of June, July, and August. This impact on neonate production was suspected of being caused by organic pesticides which are used for insect control on lawns. The effluent was never acutely toxic to C. dubia. Primarily, toxic occurrences in either the effluent or the river samples were primarily of a chronic nature. Overall, survival of C. dubia was affected more frequently at the upstream site than in the effluent or …

Carbonic anhydrase is a ubiquitous zinc-metalloenzyme that catalyzes the interconversion of carbon dioxide and carbonate and has been found to play a wide range of roles in animals, plants and bacteria. Cotton genomic and cDNA libraries were screened for the plastidial isoform of carbonic anhydrase. The nucleotide sequences of two 1.2 Kb partial cDNA clones were determined. These clones exhibit high homology to carbonic anhydrases from other dicot plants and possess all the expected peptide motifs. For example, serine and threonine rich chloroplastic targeting peptide and conserved zinc binding residues are both present. These clones were utilized to isolate two carbonic anhydrase genes that were shown to encode different isoforms by PCR and RFLP analysis.

Acute effects of a pulsed external electric field (PEEF) at 20 V/cm and a d.c. EEF at 90 V/cm on light transmittance in an isolated compound crayfish nerve was measured. In a third series, the nerve was pre-treated with the Na+ channel blocker tetrodotoxin (TTX). A PEEF produced an irreversible increase in the variation of light transmittance in normal nerves but a reversible increase in TTX treated nerves. This data was statistically insignificant. The d.c. EEFs produced a reversible and statistically significant enhancement of variation in light transmittance in both untreated and TTX-treated nerves. The findings may be due to either (1) an alteration in the ion/fluid flux within the nerve or (2) a physical alteration of protein molecules in the membranes.

This study employed a new model of mild-to-moderate head trauma to specifically identify the role of dural mast cell (MC) histamine in trauma-induced increased permeability in the blood-brain barrier (BBB). A single line was scored partially through the left dorsal parietal skull. Immediately following the trauma, degranulation was seen in 39% of the MCs on the left and in 2% on the right. After a 20 min survival period, left duras showed 55% with MC degranulation (fewer with complete degranulation) compared to 34% on the right. In the other experiments two parallel lines were scored following the injection of Evan's blue. Histamine assay showed histamine increased in the left cortex to 154% at 5 min, 174% at 10 min, and 151% at 20 min. Fluorescent quantitation of extravasated Evan's blue at 20 min following the trauma gave an increase of 1385% over the value measured for the right cortex. Zolantidine, a selective histamine H2 receptor antagonist, administered at 10- and 20- mg/kg 30 min before the trauma blocked 65% of the Evan's blue extravasation compared with the control and 2.5 mg group.

This thesis consists of one major section with two subsections. The first subsection investigates the activity of Arthrobacter globiformis aspartate transcarbamylase's specific activity with increasing concentrations of the enzyme's substrate. Dihydroorotase (DHOase) activity was also measured with increasing concentrations of the substrate dihydroorotate. The second subsection collected data in order to classify the enzyme, resulting in a classification into the category of class A ATCases with bifunctional ATCase-DHOase complexes. The thesis provides evidence to broaden understanding of the ATCase and DHOase enzymes for members of the family that Arthrobacter belongs to.

Sensilla styloconica on the proboscis of 107 species of North American and tropical butterflies were comparatively studied using the scanning electron microscope. Focus was on 76 species of North American Nymphalidae representing 45 genera and 11 subfamilies. Nomenclature for generalized and specific types of nymphalid sensilla is proposed. Written descriptions and micrographs are presented for each species studied. Morphological features were generally consistent for all or most species within genera and sometimes within subfamilies, with specified exceptions. Statistical analysis revealed significant differences for six of eight variables tested between two distinct feeding guilds of North American Nymphalidae. Average number, density, extent of proboscis coverage with sensilla, their total length, and shoulder spine length were all significantly greater in the non-nectar feeding guild than in nectar feeders, and may indicate adaptation for greater efficiency in feeding on flat surfaces. The greater frequency of apical shoulder spines in non-nectar feeders may represent adaptation for protection of sensory pegs from mechanical abrasion during feeding, or for anchoring the flexible proboscis tip to the surface. Correlation analysis revealed 9 out of 28 positive correlations in nectar feeders and 5 out of 28 in non-nectar feeders. Results of preliminary cladistic analysis were not considered to …

Estrogen is the natural agonist of the estrogen receptor (ER). However, certain plant-derived compounds or phytoestrogens have been identified that mimic estrogens and act as agonists and/or antagonists of ERs, depending on subtype and target tissue. Understanding how phytoestrogens interact with ERs, and therefore effect the estrogenic response, may prove beneficial in hormone replacement therapy and in the prevention and treatment of hormone-related diseases. Using Thin Layer Chromatography, gas chromatography/mass spectrometry (GC/MS), and proton nuclear nagnetic resonance (HNMR), I identified 4-ethoxymethylphenol (4EM) found in Maclura pomifera. While most phytoestrogens are heterocyclic compounds, 4EM is a simple phenol that acts as an agonist of ER-alpha and -beta in HeLa and MCF-7 cells. To study the effect of 4EM on ER-alpha and -beta activity, I performed transient transfection assays and showed that 4EM activates ER dependent gene transcription in a dose dependent manner in both ER subtypes. Further, 4EM- mediated transcription in ER-alpha, like estrogen, was enhance in the presense of co-activators, SRC-1 (steroid receptor coactivator-1), CBP (CREB binding proteins), and E6-AP (E6-associated protein) and inhibited by trans-4- hydroxytamoxifen (4HT). I found that 4EM was specific for ER and did not activate transcription of the progesterone receptor in HeLa cells.

This manual has been designed for a class of twenty- four students concurrently enrolled in the lecture course. The laboratory aids in the learning process and fosters an interest in the science of genetics. This manual and the experiments contained within are both informative and fun. The manual correlates with and expands upon the genetics course. Each investigation, with the exception of the Drosophila melanogaster project, can be completed in a 3-4 hour timeframe. This manual provides a “hands on” experience of theories simply discussed in the lecture course. This manual is intended to be a one-source manual where each investigation is designed to include an adequate introduction. Special attention has been given for each investigation with both the student and instructor in mind.

This thesis presents a model for developing site-specific contingency plans to be used during spill response, remediation, and post-spill monitoring using the Strategic Petroleum Reserve's Bryan Mound site as a case study. Bird and vegetation observations provide baseline data for biological conditions, and sediment sampling for total petroleum hydrocarbons serves as a chemical component of the model. Results demonstrate previously unknown conditions that would hinder remediation and affect the persistence of petroleum contaminants. Results also established previously unmapped dominant bird and vegetation types likely to be impacted by a spill at the site. This model points to a reconsideration of individual facilities' responsibilities when planning for large-scale disasters and protecting the sensitive ecosystems surrounding their sites.

Methanococcus jannaschii is a thermophilic methane producing archaebacterium. In this organism genes encoding the aspartate transcarbamoylase (ATCase) catalytic (PyrB) and regulatory (PyrI) polypeptides were found. Unlike Escherichia coli where the above genes are expressed from a biscistronic operon the two genes in M. jannaschii are separated by 200-kb stretch of genome. Previous researchers have not been able to show regulation of the M. jannaschii enzyme by the nucleotide effectors ATP, CTP and UTP. In this research project we have genetically manipulated the M. jannaschii pyrI gene and have been able to assemble a 310 kDa E. coli like enzyme. By using the second methionine in the sequence we have shown that the enzyme from this organism can assemble into a 310 kDa enzyme and that this enzyme is activated by ATP, CTP and inhibited by UTP. Thus strongly suggesting that the second methionine is the real start of the gene. The regulation of the biosynthetic pathway in Pseudomoans aeruginosa has previously been impossible to study due to the lack of CTP synthase (pyrG) mutants. By incorporating a functional uridine (cytidine) kinase gene from E. coli it has been possible to isolate a pyrG mutant. In this novel mutant we have …

The Trinity River flows through the Dallas-Ft. Worth Metroplex in north central Texas where it receives effluents from numerous point sources including seven large regional wastewater treatment facilities. Historically, the Trinity River has been impacted by massive wastewater loadings which often constitute > 80% of the total river discharge during low flow periods. Normally, high mass loadings correspond to the summer months, compounding the effects of a naturally stressful period, characterized by high temperatures and low dissolved oxygen concentrations. Samples from 12 stations were collected quarterly over an 18 month period from the Trinity River and two tributaries. Water samples were analyzed for a variety of water quality variables, including metals, priority pollutants, pesticides, and general water quality parameters. Water samples were also tested for acute and subchronic effects with several test species. Fish were collected at each station and assemblages were characterized using traditional classification techniques and the Index of Biotic Integrity. In addition, sediment samples were assessed for toxic effects which could have adversely impacted fish recruitment and in situ biomonitoring experiments were performed. Quantitative habitat characterization analyses were performed to gain additional information that could possibly explains differences in fish assemblage structure related to habitat variability. Data …

Drumming behavior is described for the first time in 33 North American Plecoptera species, and signals of an additional five species, Pteronarcys pictetii, Acroneuria abnormis, Paragnetina media, Clioperla clio and Isogenoides zionensis, are further detailed. An out-group comparison of behavioral characters in all 104 world species whose drumming is known showed that the behavior is more advanced in the Arctoperlaria Group Systellognatha than in the Group Euholognatha. In general, tapping, monophasy, touching, sequenced exchange and less than 50 taps/answer are ancestral states, and rubbing, grouping, phasing, tremulation, interspersed exchange and equal or more than 50 taps/answer are derived states. There has been some co-evolution between abdominal structure and drumming behavior. Scanning Electron Micrographs of 30 species showed that the primitive state of tapping is ascociated with three classes of abdominal structure: (1) absence of derived structures, (2) lobes or vesicles, and (3) hammers. The derived behavior of rubbing, however, occurs only in species with derived structures, and is predominant in species having vesicles and hammers. Drumming can be used as a line of evidence to aid in defining genera and species, since the behavior has a variable degree of specificity or exclusiveness in all species, particularly in groups of species …

Cyanide is a well known toxicant that arises in the environment from both biological and industrial sources. Bacteria have evolved novel coping mechanisms for cyanide and function as principal agents in the biosphere for cyanide recycling. Some bacteria exhibit the unusual ability of growing on cyanide as the sole nitrogen source. One such organism is Pseudomonas fluorescens NCIMB 11764 (Pf11764) which employs a novel oxidative mechanism for detoxifying and assimilating cyanide. A unique complex of enzymes referred to as cyanide oxygenase (CNO) is responsible for this ability converting cyanide to ammonia which is then assimilated. Because one component of the four member CNO complex was previously shown to act on cyanide independent of the other members, its characterization was sought as a means of gaining a better understanding of the overall catalytic mechanism of the complex. Preliminary studies suggested that the enzyme belonged to a subset of nitrilase enzymes known as cyanide dihydratases (CynD), however, a cynD-like gene in Pf11764 could not be detected by PCR. Instead, a separate nitrilase (Nit) linked to cyanide metabolism was detected. The corresponding nit gene was shown to be one of a conserved set of nit genes traced to a unique cluster in bacteria …