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  Partner: UNT Libraries
 Department: Department of Biological Sciences
 Degree Level: Master's
 Collection: UNT Theses and Dissertations
N-Acylethanolamine (NAE) Profiles Change During Arabidopsis Thaliana Seed Germination and Seedling Growth.
An understanding of the potential roles as lipid mediators of a family of bioactive metabolites called N-acylethanolamines (NAEs) depends on their accurate identification and quantification. The levels of 18C unsaturated NAEs (e.g. NAE18:2, NAE 18:3, etc.) in wild-type seeds (about 2000 ng/g fw) generally decreased by about 80% during germination and post-germinative growth. In addition, results suggest NAE-degradative fatty acid amide hydrolase (FAAH) expression does not play a major role in normal NAE metabolism as previously thought. Seedlings germinated and grown in the presence of abscisic acid (ABA), an endogenous plant hormone, exhibited growth arrest and secondary dormancy, similar to the treatment of seedlings with exogenous N­lauroylethanolamine (NAE12:0). ABA-mediated growth arrest was associated with higher levels of unsaturated NAEs. Overall, these results are consistent with the concept that NAE metabolism is activated during seed germination and suggest that the reduction in unsaturated NAE levels is under strict temporal control and may be a requirement for normal seed germination and post-germinative growth.
N-Acylethanolamines and Plant Phospholipase D
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.
Adaptive Advantages of Carotenoid Pigments in Alpine and Subalpine Copepod Responses to Polycyclic Aromatic Hydrocarbon Induced Phototoxicity
Alpine zooplankton are exposed to a variety of stressors in their natural environment including ultraviolet radiation. Physiological coping mechanisms such as the accumulation of photoprotective compounds provide these zooplankton protection from many of these stressors. Elevated levels of carotenoid compounds such as astaxanthin have been shown to help zooplankton survive longer when exposed to ultraviolet radiation presumably due to the strong antioxidant properties of carotenoid compounds. This antioxidant capacity is important because it may ameliorate natural and anthropogenic stressor-induced oxidative stress. While previous researchers have shown carotenoid compounds impart increased resistance to ultraviolet radiation in populations of zooplankton, little work has focused on the toxicological implications of PAH induced phototoxicity on zooplankton containing high levels of carotenoid compounds. This thesis discusses research studying the role that carotenoid compounds play in reducing PAH induced phototoxicity. By sampling different lakes at elevations ranging from 9,500' to 12,700' in the front range of the Colorado Rocky Mountains, copepod populations containing different levels of carotenoid compounds were obtained. These populations were then challenged with fluoranthene and ultraviolet radiation. Results discussed include differences in survival and levels of lipid peroxidation among populations exhibiting different levels of carotenoid compounds, and the toxicological and ecological implications of these results.
Adherence and Haemagglutination of Moraxella Catarrhalis.
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M. catarrhalis is a gram-negative diplococci frequently associated with infections of the upper respiratory tract. During the past decade, some preliminary studies have attempted to elucidate mechanisms of adherence and haemagglutination of M. catarrhalis. These studies have reported, in many cases, inconsistent results. There are two purposes of this research. First, identify mechanisms that may potentially be associated with the adherence and haemagglutination of M. catarrhalis. Second, suggest research directions that may be fruitful in clarifying these mechanisms.
Alterations in Fatty Acid Amide Hydrolase (Faah) Transcript Levels and Activity Lead to Changes in the Abiotic Stress Susceptibility of Arabidopsis Thaliana
N-Acylethanolamines (NAEs) are a class of bioactive lipids, and FAAH is one of the enzymes responsible for degrading NAEs in both plants and animals. in plants, FAAH appears to be closely associated with ABA, a phytohormone which has long been associated with plant stress responses, since the overexpression of FAAH in Arabidopsis results in ABA hypersensitivity. Therefore, it is reasonable to speculate that alterations in FAAH transcript levels will result in altered stress responses in plants. to investigate this hypothesis experiments were carried out in which wild type (WT), FAAH-overexpressing (OE), and T-DNA insertional FAAH knockouts of Arabidopsis (faah) were grown in MS media under stress conditions. the stress conditions tested included chilling stress, heavy metal stress induced by cadmium or copper, nutrient limitations induced by low phosphorus or low nitrogen, salt stress induced with NaCl, and osmotic stress induced with mannitol. the OE plants were consistently hypersensitive to all stress conditions in relation to wild type plants. Inactive FAAH overexpressors did not have the hypersensitivity to the salt and osmotic stress of the active OE plants and were instead tolerant to these stresses. FAAH2 (faah2) knockouts and FAAH 1 and 2 double knockouts (faah 1+2) were based on some root development parameters somewhat more tolerant than WT plants, but more sensitive in terms of shoot growth. Collectively the data suggests that FAAH activity may interact with stress-responsive pathways in plants, perhaps including pathways involving ABA.
Analysis and Development of Post Secondary Curriculum on Sustainability
This thesis examines existing curricula at colleges and universities about sustainability and uses results to develop an introductory post secondary course curriculum. The proposed course is organized around three major elements - - science, philosophy, and economics - - all integral to understanding sustainability. Materials needed to teach the proposed 3-semester hour course including syllabus, teaching modules, transparencies, handouts, and exams were developed. Suggestions on how to teach a one-semester hour course on sustainability and a workshop on sustainability are also presented. The following research and curriculum development was a project established and funded by the Texas Energy Office, Renewable Resources and Sustainability Program.
Analysis and expression of the cotton gene for the D-12 fatty acid desaturases 2-4 (FAD2-4)
A genomic clone containing a 16.9-kb segment of cotton DNA was found to encompass a D-12 fatty acid desaturases (FAD2-4) gene. The FAD2-4 gene has a single, large intron of 2,780 bp in its 5'-untranslated region, just 12 bp upstream from the ATG initiation codon of the FAD2-4 opening reading frame. A number of prospective promoter elements, including several light-responsive sequences, occur in the 5'-flanking region. The coding region of the gene is 1155 bp with no introns, and would encode a FAD2-4 polypeptide of 384 amino acids. The putative protein had four membrane-spanning helices, hallmarks of an integral membrane protein, and would probably be located in the endoplasmic reticulum. The FAD2-4 gene is indeed a functional gene, since yeast cells transformed with a plasmid containing the coding region of the gene synthesize an appreciable amount of linoleic acid (18:2), not normally made in wild-type yeast cells. The FAD2-4 gene has many structural similarities to the cotton FAD2-3 gene that was also analyzed in this laboratory.
Analysis of the One-Horned Rhinoceros (Rhinoceros Unicornis) Habitat in the Royal Chitwan National Park, Nepal.
This study analyzes the remaining suitable habitat of the one-horned rhinoceros, Rhinoceros unicornis, in Royal Chitwan National Park of Nepal. An April 2003 Landsat image was classified into eight land cover types: wetland, sand, water, mixed forest, sal forest, agriculture, settlement, and grassland. This image was converted into habitat suitability maps using cover, food, and water. The rhinoceros prefers grassland habitat with oxbow lakes and closed canopy during the monsoon season. Nominal values of five parameters were used to create a map of habitat suitability index. The map was categorized into four habitat classes: highly unsuitable, unsuitable, moderately suitable habitat, and suitable. Landscape metrics, patch metrics and class metrics associated with habitat were determined through the use of FRAGSTATS.
Anatomical and Morphological Responses of Cardiospermum Halicacabum L. (Balloon Vine), to Four Levels of Water Availability
C. halicacabum (Sapindaceae) is an invasive plant that is considered a nuisance species in Texas riparian environments. Little is known of the tolerance of C. halicacabum to flooding and drought; however, this information may provide insight into the characteristics that contribute to C. halicacabum purported invasiveness. C. halicacabum seedlings (n = 92) were exposed to one of four levels of water availability (flooded, saturated, intermediate and dry) over six weeks under greenhouse conditions. Plant performance was affected by water availability; however, there was no effect on survivorship. Flooded and saturated plants exhibited morphological adaptations; producing adventitious roots, hypertrophy, and aerenchyma tissue. Morphological measures, anatomical responses, and patterns of biomass allocation all indicate that C. halicacabum is able to survive periodic inundation, perform in saturation, and establish and thrive on the drier end of a moisture gradient.
Animal Contribution to Human Medicine
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The use of animal models in research has led to a fierce debate between animal rights activists and scientists. The former claim that little useful information is gained from animal studies and the suffering of animals does not preclude any treatments which may be used to treat human illnesses. Yet, research scientists claim that in vivo animal models are of absolute necessity to developing treatments and cures to disease. To determine the necessity of animal use, one must examine the models currently in research. Have the animal models for disorders such as cystic fibrosis and muscular dystrophy given scientists enough information to develop effective treatments? This paper will examine the role of animal subjects in several disease research protocols, as well as the applicability of the research.
Applications of Remote Sensing and GIS to Modeling Fire for Vegetative Restoration in Northern Arizona
An accurate fire model is a useful tool in predicting the behavior of a prescribed fire. Simulation of fire requires an extensive amount of data and can be accomplished best using GIS applications. This paper demonstrates integrative procedures of using of ArcGIS™, ERDAS Imagine™, GPS, and FARSITE© to predict prescribed fire behavior on the Kaibab-Paiute Reservation. ArcGIS was used to create a database incorporating all variables into a common spatial reference system and format for the FARSITE model. ArcGIS Spatial Analyst was then used to select optimal burn sites for simulation. Our predictions will be implemented in future interagency efforts towards vegetative restoration on the reservation.
The aquatic insect communities of Holbrook Creek and Cochetopa Creek in Colorado.
The first objective for this problem in lieu of thesis project was to gather, identify to the lowest practical taxonomic level and organize all available aquatic insects collected from high altitude Colorado aquatic systems during the summers of 1994, 1996, 1998, and 2002 for the University of North Texas Environmental Science Field Course (BIOL 5650). The curated collection will be housed in the Elm Fork Natural History Museum, located at the University of North Texas. The second objective was to provide a summary and discussion of the occurrence and distribution of the aquatic insects collected from Mt. Blanca in 1994, 1996, and 1998 and to create a taxa list of aquatic insects collected from Cochetopa Creek during the summer of 2002.
Aspartate Transcarbamoylase of Aeromonas Hydrophila
This study focused on the enzyme, aspartate transcarbamoylase (ATCase) from A. hydrophila, a Gram-negative bacterium found in fresh water. The molecular mass of the ATCase holoenzyme from A. hydrophila is 310 kDa. The enzyme is likely composed of 6 catalytic polypeptides of 34 kDa each and 6 regulatory polypeptides of 17 kDa each. The velocity-substrate curve for A. hydrophila ATCase is sigmoidal for both aspartate and carbamoylphosphate. The Km for aspartate was the highest to date for an enteric bacterium at 97.18 mM. The Km for carbamoylphosphate was 1.18 mM. When heated to 60 ºC, the specific activity of the enzyme dropped by more than 50 %. When heated to 100 ºC, the enzyme showed no activity. The enzyme's activity was inhibited by ATP, CTP or UTP.
Assessing Effects of an Environmental Education Field Science Program Fostering Responsibility at an Urban Middle School
The study investigated the ability of an extracurricular program to influence environmental responsibility of sixth and seventh graders. The Children's Environmental Attitude and Knowledge Survey (CHEAKS) was evaluated for appropriateness in assessing the worth of this particular environmental education strategy emphasizing water quality fieldwork and technology. CHEAKS is designed with psychometric reliability and validity that may be used in comparing disparate programs. Wilcoxon two sample tests were used to analyze data gathered from two student groups; one participated in an "Enviro-Mentals Club"; the other received no treatment. Analysis showed no significant change in environmental attitudes between groups, but did show significance (p <= 0.05) in environmental knowledge growth. Therefore, the investigated program had marginal success in influencing environmental responsibility.
Assessing Outcomes of a Recycling Education and Service Program within an Elementary School
During the spring 2004 a pilot school recycling program was implemented within Robert E. Lee Elementary. The primary goal of the program was to determine how recycling education in the school would affect curbside recycling rates within the surrounding community. The program was a cooperative effort between the University of North Texas, City of Denton Solid Waste Department and Keep Denton Beautiful. Throughout the first months of the study during the spring 2004, an increase in curbside recycling within the Robert E. Lee Elementary attendance zone was observed, with a dramatic decrease in participation over the summer and a rapid increase once again during the second full semester of the study. In a survey conducted with 3rd and 5th grade students at the pilot project school, most students expressed positive attitudes about recycling. Students whose survey responses indicated a high level of knowledge about what could be recycled were 37% more likely to claim to recycle regularly, than those students that scored low on the knowledge portion of the survey. Although the total amount of waste generation (recyclable and non-recyclable) at Robert E. Lee Elementary did not decrease during the study, the campus was able to divert recyclable material from their trash at a much higher rate than two other local elementary campuses with paper-only recycling and no associated recycling education program. Based upon the success of the recycling program at Robert E. Lee Elementary, the City of Denton Recycling Division has agreed to move forward with offering recycling to more schools within the Denton Independent School District during the 2005-2006 school year.
Assessing the Effects of a Municipal Wastewater Treatment Plant Effluent on Zooplankton, Phytoplankton and Corbicula Flumina in a Constructed Wetland
Wetland wastewater treatment offers low-cost, energy efficient alternatives to conventional wastewater technologies. In this study, an artificial wetland was constructed at the City of Denton, Texas Pecan Creek Water Reclamation Plant to facilitate diazinon removal from treated effluent.
Assessing the Spatial and Temporal Distribution of MTBE and BTEX Compounds in Lake Lewisville, Texas February 1999 - February 2000
The spatial and temporal distribution of Methyl Tertiary-Butyl Ether (MTBE) and BTEX (Benzene, Toluene, Ethylbenzene, Xylenes) compounds were assessed in a multipurpose reservoir, Lake Lewisville, Texas between February 1999 and February 2000. Concentrations of MTBE ranged from 0.0 - 16.7 mg/L. Levels of MTBE in the lake were related to watercraft. BTEX concentrations were never detected above 2.0 mg/L during the sampling period. Finished drinking water from Denton and the Upper Trinity Regional Water District (UTRWD) Treatment Plants were also tested for MTBE and BTEX. MTBE and BTEX were not detected in UTRWD water samples. Denton's finished water samples never exceeded 2.2 mg/L for MTBE and BTEX was not detected except for one replicate of 1.1 mg/L toluene.
An Assessment of Storm Water Toxicity from the Dallas/Fort Worth Metroplex and Denton, Texas
With the advent of national storm water regulations, municipalities with populations greater than 100,000 are required to obtain National Pollutant Discharge Elimination System Permits (NPDES) for storm water discharges. In addition to the sampling required for the permit process, the City of Fort Worth contracted with the University of North Texas' Institute of Applied Sciences to conduct acute toxicity testing using Pimephales prcmelas and Ceriodaphnia dubia on storm water samples received from the Dallas/Fort Worth Metroplex. A Toxicity Identification Evaluation (TIE) was performed on four samples that exhibited acute toxicity to C. dubia. High levels of metals as well as diazinon were some of the probable toxicants found.
Attenuation of Escherichia Coli Aspartate Transcarbamoylase Expressed in Pseudomonas Aeruginosa Mutant and Wild Type Strains
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.
Bacterial Challenge in Lumbricus Terrestris: A Terrestrial Invertebrate Immunotoxicity Model.
A bacterial challenge assay was developed utilizing the earthworm, Lumbricus terrestris, in order to assess potential immunotoxic effects from exposure to specific polychlorinated biphenyl congeners. Earthworms were inoculated with Aeromonous hydrophila, establishing a 10-day LD50. In vitro assays for effects of PCBs on phagocytosis agreed with mammalian studies, demonstrating potent suppression of phagocytosis by the non-coplanar PCB congener 138 and no suppression by the coplanar congener 126. However, when the effects of the two PCB congeners were evaluated for suppression of resistance to a whole animal infection challenge assay, coplanar PCB 126 decreased the ability of L. terrestris to withstand infection while non-coplanar PCB 138 did not.
Bacterial Cyanide Assimilation: Pterin Cofactor and Enzymatic Requirements for Substrate Oxidation
Utilization of cyanide as the sole nitrogen source by Pseudomonas fluorescens NCIMB 11764 (Pf11764) occurs via oxidative conversion to carbon dioxide and ammonia, the latter satisfying the nitrogen requirement. Substrate attack is initiated oxygenolytically by an enzyme referred to as cyanide oxygenase (CNO), which exhibits properties of a pterin-dependent hydroxylase. The pterin requirement for Pf11764 CNO was satisfied by supplying either the fully (tetrahydro) or partially (dihydro) reduced forms of various pterin compounds at catalytic concentrations (0.5 µM). These compounds included, for example, biopterin, monapterin and neopterin, all of which were also identified in cell extracts. A related CNO-mediated mechanism of cyanide utilization was identified in cyanide-degrading P. putida BCN3. This conclusion was based on (i) the recovery of CO2 and NH3 as enzymatic reaction products, (ii) the dependency of substrate conversion on both O2 and NADH, and (iiii) utilization of cyanide, O2 and NADH in a 1:1:1 reaction stoichiometry. In contrast to findings reported for Pf11764, it was not possible to demonstrate a need for exogenously added pterin as a cofactor for the PpBCN3 enzyme system. However, results which showed that cells of PpBCN3 contained approximately seven times the amount of pterin as Pf11764 (of which a significant portion was protein-bound) were interpreted as indicating that sufficient bound CNO-cofactor exists, thus eliminating any need for a supplemental source.
A Behavioral Model for Detection of Acute Stress in Bivalves
A behavioral model for acute responses in bivalves, was developed using time series analysis for use in a real-time biomonitoring unit. Stressed bivalves closed their shell and waited for the stressful conditions to pass. Baseline data showed that group behavior of fifteen bivalves was periodic, however, individuals behaved independently. Group behavior did not change over a period of 20 minutes more than 30 percent, however, following toxic exposures the group behavior changed by more than 30 percent within 20 minutes. Behavior was mathematically modeled using autoregression to compare current and past behavior. A logical alarm applied to the behavior model determined when organisms were stressed. The ability to disseminate data collected in real time via the Internet was demonstrated.
Beta-adrenergic Blockade Via Atenolol and Its Effects on Blood Pressure, Heart Rate, and Renal Morphology in the Developing Chicken Gallus Gallus Domesticus
Chicken embryos were chronically exposed to the ?1- blocker atenolol during one of three stages: mesonephros (E7-E9), mesonephros-metanephros (E11-E13), or metanephros (E15-E17). Mesonephros group hearts were larger than all other groups (P < 0.01). Mesonephros and metanephros group kidneys were larger than all remaining groups (P < 0.0001). The mesonephros group nephron number was ~40% lower than control values (P = 0.002). Glomerular areas were 26% and 18% larger than the control group in the mesonephros and metanephros groups, respectively (P < 0.001). These data suggest an E7-E9 critical window of cardiovascular and renal development for atenolol. Acute atenolol exposure in E15 embryos showed an increase in mean arterial pressure with all but the highest dose. All doses significantly decreased heart rate.
Bioconcentration and Morphological Effects of Triclosan on Three Species of Wetland Plants
Triclosan (TCS) is an antimicrobial compound found in several types of common household products. After being washed down the drain, TCS will then end up in the local watershed. Although numerous studies have been conducted to evaluate the fate and effects of TCS in aquatic environments, there have been no studies evaluating the role arbuscular mycorrhizal fungi (AM) play in a plants response to TCS exposure. Three species of wetland plants native North Texas were inoculated with AM spores and exposed to 0, 0.4 g/L and 4.0 g/L TCS concentrations. Root morphology of E. prostrata and S. herbacea showed AM and exposure responses. S. herbacea produced the greatest amounts biomass and TCS bioaccumulation, in all but one treatment. It also displayed opposing results to E. prostrata in measures of root length, root surface area, relative root mass, relative shoot mass and shoot:root ratio. TCS root tissue concentrations increased with increased exposures for both E. prostrata and S. herbacea. Even though E. prostrata had the lowest levels in each measure of biomass production, it had the highest amount of root TCS bioaccumulation in the AM inoculated 4.0 g/L treatment. H. laevis was between the other two species in terms of biomass production, and did not demonstrate any exposure or inoculation effects in morphology or TCS accumulation. Overall, no clear patterns were detected, which highlights the fact that further study is required to completely understand the effects compounds like TCS can have on plant community structure, and ultimately ecosystem function.
Bioconcentration of Triclosan, Methyl-Triclosan, and Triclocarban in the Plants and Sediments of a Constructed Wetland
Triclosan and triclocarban are antimicrobial compounds added to a variety of consumer products that are commonly detected in waste water effluent. The focus of this study was to determine whether the bioconcentration of these compounds in wetland plants and sediments exhibited species specific and site specific differences by collecting field samples from a constructed wetland in Denton, Texas. The study showed that species-specific differences in bioconcentration exist for triclosan and triclocarban. Site-specific differences in bioconcentration were observed for triclosan and triclocarban in roots tissues and sediments. These results suggest that species selection is important for optimizing the removal of triclosan and triclocarban in constructed wetlands and raises concerns about the long term exposure of wetland ecosystems to these compounds.
Biology and Energetics of Tropisternus Lateralis Nimbatus (SAY) (Coleoptera: Hydrophilidae) in a Playa on the Southern High Plains of Texas and Aquatic Coleoptera Diversity from Seven Playas on the Southern High Plains of Texas
A study of the biology of Tropisternus lateralis, a hydrophilid beetle, was conducted during the flood period of a single playa on the Southern High Plains of Texas from early June 1995 through early September 1995. Mechanism of colonization, tolerance/avoidance to drought, larval density, and secondary production were analyzed. T. lateralis colonized playas from surrounding aquatic habitats and avoided drought through aerial dispersion. Once in the playa, larval density increased over time. Secondary production was 1.31 g/m2/.25 yr. In addition, aquatic Coleoptera diversity was studied in seven playas on the Southern High Plains of Texas. A total of twenty three species were identified from the study region. Nine species not reported in playa literature were identified.
Bioreactor Landfill Cell Feasibility Study ' Reference to City of Denton Subtitle-D Permit #1590A Landfill
The City of Denton Landfill, Permit #1590A, utilizes “Dry-Tomb” techniques for disposal and promotion of municipal solid waste stabilization, as described by the Resource Conservation and Recovery Act (RCRA) prohibition in 40 CFR. Bioreactor research suggests re-circulating leachate increases biodegradation rates and reduces long-term monitoring from fifty years to less than ten years. Current procedures that are followed at Denton's landfill, literature review and the use of the Hydrologic Evaluation of Landfill Performance (HELP) model, suggest that a bioreactor landfill cell is worthy of further research. Re-circulating leachate and augmenting it with additional liquid will increase biodegradation and the need to design and build a landfill gas collection system to capture methane for energy recovery uses.
Biotic Factors and Temperature Tolerances via Critical Thermal Methodology in Goldfish
CTMinimum and CTMaximum were measured in 620 goldfish to determine if biotic factors, in particular starvation, condition factor and size, affect temperature tolerance. Twenty-eight days of starvation adversely affected both upper and lower temperature tolerance. Condition factor was related to upper but not lower temperature tolerance.
A Carcinogenic Agent Elaborated by Liver Cells from Lymphosarcoma-Bearing Mice
Liver cells from lymphosarcoma-bearing DBA/1J mice were shown, by parabiotic culture with normal liver cells from isologous mice, to elaborate an agent which could pass a 25 mu filter and transform the normal cells to a malignant state.
Cassette Systems for Creating Intergeneric Hybrid ATCases
Cassette systems for creating intergeneric hybrid ATCases were constructed. An MluI restriction enzyme site was introduced at the carbamoylphosphate binding site within the pyrB genes of both Pseudomonas putida and Escherichia coli. Two hybrids, E. coli pyrB polar domain fused with P. putida pyrB equatorial domain and P. putida pyrB polar domain fused with E. coli pyrB equatorial domain, are possible. The intergeneric E. coli-P. putida hybrid pyrB gene was constructed and found to encode an active ATCase which complemented an E. coli Pyr- strain. These hybrids are useful for kinetic and expression studies of ATCase in E. coli.
Characterization of Arthrobacter Globiformis Aspartate Transcarbamylase Concentrations of Substrates
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.
Characterization of Aspartate Transcarbamoylase and Dihydroorotase in Moraxella Catarrhalis
Bacterial aspartate transcarbamoylases (ATCase's) are divided into three classes that correspond to taxonomic relationships within the bacteria. The opportunistic pathogen Moraxeila catarrhalis has undergone several reclassifications based on traditional microbiological criteria. The previously uncharacterized ATCase from M. catarrhalis was purified to homogeneity and its chemical properties characterized. The ATCase from M. catarrhalis is a class C ATCase with an apparent molecular mass of 480-520 kDa. The M. catarrhalis ATCase is a dodecomer composed of six 35 kDa polypeptides and six 45 kDa polypeptides. The enzyme has an unusually high pH optimum of greater than pH 10. The enzyme exhibited hyperbolic kinetic with a Km for aspartate of 2 mM. A single, separate 78 kDa dihydroorotase from M. catarrhalis was identified and it was not associated with ATCase. These data support the reclassification of M. catarrhalis out of the Neisseriaceae family.
A Characterization Of Jackson Blue Spring, Jackson County, Florida
Jackson Blue is a first magnitude spring in the karst terrane of northeast Florida. Previous studies have identified inorganic fertilizer as the source of high nitrate levels in the spring. Agricultural land use and karst vulnerability make Jackson Blue a good model for conservation concerns. This work offers an aggregation of studies relating to the springshed, providing a valuable tool for planning and conservation efforts in the region. An analysis of nitrate levels and other water quality parameters within the springshed did not reveal significantly different values between agricultural and forested land use areas. Confounding factors include: high transmissivity in the aquifer, interspersed land use parcels, and fertilizer application in forested areas due to commercial pine stand activity.
Characterizing Spontaneous Neurophysiological Activity with Measures of Statistical Serial Dependence: a Summary Statistic and an Extension of the Joint Interval Histogram
Two measures which indicate statistical serial dependence were evaluated. The n-dimensional Average Stored Information index (ndASI) is a measure of conditional information, which compares the entropies of higher order conditional distributions to estimate average statistical serial dependence. The generalized ranked joint interval histogram (RJIH-o) is a new nonparametric graphical analysis method. It extends the joint interval histogram by depicting longer interval sequences and can be interpereted precisely analagously to the joint interval histogram. The generalized ranked joint interval histogram correctly represents independence in a Poisson process model and statistical serial dependence in a directionally reinforced Markov model. The generalized ranked joint interval histogram correctly depicts the underlying periodic and strange attractors in a standard map model. Both measures can be used to effectively analyze interspike interval sequences from spontaneous neurophsyiological activity effectively.
Characterizing the Municipal Solid Waste Stream in Denton, Texas
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Forty-two commercially collected dumpsters from Denton’s Municipal Solid Waste Stream were emptied, sorted and weighed to characterize the material types and make preliminary recycling policy recommendations. The general composition of Denton’s solid waste stream was not significantly different from the composition of the nation’s solid waste stream. Fifty-eight percent of the observed waste stream was recyclable. Paper made up the largest portion of recyclable materials and the "grocery" source category had more paper than any of the other five categories. Based on these findings, an incrementally aggressive approach is recommended to reduce certain types of wastes observed in the waste stream. This would include a Pay-As-You-Throw Program followed by an Intermediate Processing Center that can be converted to a Materials Recovery Facility.
Chemotactic Response of Lumbricus terrestris Coelomocytes to Larval and Adult Stages of Rhabditis pellio
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Experiments were performed to assess the suitability of Rhabditis pellio, a nematode found in earthworms, as a challenge organism for use in development of a biomarker assay to determine the potential of chemicals to suppress the immunocompetence of the non-specific immune system. To accomplish this goal, information on the life cycle of R. pellio was determined; including effects of incubation time and temperature on growth rates; along with information on the immune response elicited in the earthworm, Lumbricus terrestris. Immune parameters measured were coelomocyte migration toward and attachment to R. pellio larvae and adults. Preliminary background information showed that R. pellio has potential as a challenge organism for development of a biomarker assay.
Chronic Hypoxia and Hyperoxia Modifies Morphology and Vegf Expression of the Lungs of the Developing Chicken (Gallus Gallus Domesticus)
This study determines effects of oxygen levels on morphology and VEGF expression of developing chicken lungs following incubation in normoxia (21% O2), hypoxia (15% O2) or hyperoxia (30% O2), until developmental days 16 or 18. Lung morphology was assessed using light microscopy, while VEGF expression was determined with ELISA. In hypoxia, the proportion of parabronchial tissue and parabronchi including lumina increased from day 16 to 18 (61 to 68% and 74.2 to 82.2%, respectively). Non-parabronchial tissue was higher in hypoxia than in hyperoxia on day 16 (26 to 20%). However, by day 18, there were no differences between groups. VEGF expression was 33% higher in hypoxia than in hyperoxia on day 16 (736 vs. 492 pg/ml). On day 18, VEGF expression was 43% higher in hyperoxia than in normoxia (673 to 381pg/ml), and remained elevated by 40% in hypoxia over normoxia (631 pg/ml). VEGF may be a mechanism by which parabronchial tissue is stimulated from day 16 to 18 following exposure to chronic hypoxia.
Classification of Toolmark Surfaces on Zipper Teeth
This study proposes the classification of the toolmark under the heads of zipper teeth as a subclass characteristic as outlined by the Association of Firearm and Toolmark Examiners (AFTE). Two separate cases in which zipper teeth were found at crime scenes prompted this study. Brass zipper teeth manufactured by YKK were taken from 20 pairs of jeans and studied using a Reichert comparison microscope at 4X power. Photographs were taken and over 750 comparisons made. It was found that the toolmarks on each side on the 20 zippers were unique and independent of all other sides. The observations made in this study indicate that classifying zipper teeth toolmarks as a subclass characteristic is valid.
Cloning of Carbonic Anhydrase from Cotton (Gossypium hirsutum L.)
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.
Clustering Algorithms for Time Series Gene Expression in Microarray Data
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.
Coelomic Fluid Protein Profile in Earthworms Following Bacterial Challenge.
Proteomic techniques were used to evaluate the protein profile of the earthworm, (Lumbricus terrestris), following a bacterial challenge. One control group received no injection; a second control group received injections of phosphate buffer solution (PBS). The experimental group received injections of PBS containing (Aeromonas hydrophila). After incubation for 12 hours at 20°C, coelomic fluid was collected from each group for analysis by 2-D electrophoresis. There were significant differences in spot appearance and density between control and experimental groups. Sixteen spots showed a two-fold increase in density and 63 showed at least a two-fold decrease in density between samples from control and bacteria-challenged earthworms, respectively, suggesting up- and down-modulation of proteins potentially involved in the earthworm's response to bacterial challenge.
College Freshman Biology Two Semester Course: Integrating Deep Processing Teaching Techniques
Development of a college level freshman biology course was undertaken in response to government reports that American students have fallen behind students of other countries in the area of the sciences. Teaching strategies were investigated to accomplish two objectives, to define essential academic material to include in the course and to investigate teaching techniques that would increase deep processing of the information. An active process that consisted of applying the cognitive information to solving problems or developing answers to questions was defined as critical thinking. Critical thinking was incorporated into the course by the use of case studies.
Comparative Bioavailability of Dietary and Dissolved Cadmium to Freshwater Aquatic Snails
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Heavy metal bioaccumulation in aquatic organisms may occur through direct or indirect uptake routes. Research indicates that the significance of uptake route varies with contaminant and organism exposed. The relative importance of different metal sources in aquatic systems was investigated by exposing freshwater snails to dietary or dissolved sources of cadmium. Snails were exposed to control, contaminated food only, contaminated water only, and contaminated food and water treatments. During the 15-day exposure, samples were taken to determine Cd concentration in snail soft tissue, snail shell, algal food, and overlying water. Analyses of snail soft tissue and shells indicate that exposure route significantly affects Cd concentrations in the tissues. In both cases, dissolved Cd is the primary contributor to metal body burden.
Comparative Mitochondrial DNA Sequence Diversity in Isolated and Open Populations of Southern Flying Squirrels
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Three populations of Southern flying squirrels were studied in the Ouachita Mountains of Arkansas to assess the impact of population subdivision-due to island formation--on the population genetics of Glaucomys volans. One island, one mainland, and one open population were investigated. A 367 nucleotide hypervariable region of mitochondrial DNA was sequenced in individuals from each population. Individuals and populations were compared to assess relatedness. Higher sequence diversity was detected in the open and island populations. One island individual shared characters with both the island and mainland populations. Results support the hypothesis that the mainland population may have reduced gene flow. Also, the island population may have been originally founded by at least two maternal lineages.
Comparative Phyto-uptake Across Distribution Coefficients of Pharmaceutical Compounds and Aquatic Macrophytes: Carbamazepine and Amiodarone Uptake in Lemna Spp
Few studies have been conducted on the effectiveness of phytoremediation of pharmaceutical compounds, although the persistent and non-acutely toxic nature of many of these compounds in today's water bodies may yield an ideal application for this practice. To quantify the potential effectiveness of plant uptake, kinetic and proportional bioconcentration factors (BCFk, and BCFp, respectively) in nanograms (ng) carbamazepine and amiodarone per gram (g) wet weight plant tissue for Lemna spp. were determined utilizing a 14-day continuous flow-through study. Samples were analyzed using isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) running in positive ion mode. Kinetic BCF was estimated at 0.538, while proportional BCF was estimated at 0.485. Kinetic BCF for the amiodarone study was estimated at 23.033, whereas proportional BCF was estimated at 41.340. Possible contamination of the C18 column and peristaltic pump failure may have impacted uptake results. In light of variability and current lack of research in the field, this work should be considered exploratory rather than conclusive.
Comparison of Aspartate Transcarbamoylase and Pyrimidine Salvage in Sporosarcina urea, Sprolactobacillus inulinus, Lactobacillus fermentum, and Micrococcus luteus
The enzyme that catalyzes the committed step in pyrimidine biosynthesis, aspartate transcarbamoylase, has been compared in selected endospore-forming organisms and in morphologically similar control organisms. The ATCases and pyrimidine salvage from Sporosarcina ureae, Sporolactobacillus inulinus, Lactobacillus fermentum, and Micrococcus luteus were compared to those of Bacillus subtilis. While the ATCases from Sporosarcina ureae, Sporolactobacillus inulinus, and L. fermentum were found to exhibit characteristics to that of Bacillus with respect to molecular weight and kinetics, M. luteus ATCase was larger at approximately 480 kDa. Furthermore, pyrimidine salvage in Sporosarcina ureae and M. luteus was identical to those of B. subtilis, while pyrimidine salvage of Sporolactobacillus inulinus and L. fermentum resembled that of the pseudomonads.
Comparison of GPS Point Selection Methods for GIS Area Measurement of Small Jurisdictional Wetlands
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U.S. Army Corps of Engineers (USACE) regulates fill of jurisdictional waters of the United States including wetlands. Recent USACE regulations set a threshold of impacts to wetlands at one-half acre. Impact area can be determined by Global Positioning System (GPS) measurement of wetland boundary and Geographic Information System (GIS) calculation of impact area. GPS point selection methods include (1) equal time interval, (2) transect and (3) intuition. Four two-acre shapes were measured with each GPS method and brought into GIS for area calculation. Analysis of variance and Root Mean Square Error analyses determine that the transect method is an inferior point selection method in terms of accuracy and efficiency.
Comparison of Heteranthera Dubia (Jacq.) MacM.-associated Macroinvertebrates Between Georgraphical Regions in the United States
Macroinvertebrates associated with the aquatic plant, water stargrass (Heteranthera dubia), were sampled from 12 waterbodies in four regions of the United States from June to August 2005. Taxa richness, evenness, and diversity were lowest in the Lower Midwest (LMW) region, and higher in Northern sites, especially the Upper Midwest (UMW), and Northeast (NE). While relative abundance varied from site to site and region to region, utilization of the plant by functional groups remained fairly constant. Collector-gatherers consistently comprised the largest portion of invertebrates sampled. The shredder/ herbivore functional group comprised an average of 17 % of total groups. Through an exhaustive literature review, it was found that shredder/ herbivores of water stargrass have not been reported in the literature. Because of this, the herbivore group was analyzed separately and consisted of 2,383 specimens representing 23 species. The most common groups were Rhopalosiphum sp., Nectopsyche spp. and chironomids. No differences were found in herbivore diversity or evenness between sampling regions, but species richness was significantly different.
Comparison of Remediation Methods in Different Hydrogeologic Settings Using Bioplume II
A contaminant fate and transport computer model, Bioplume II, which allows simulation of bioremediation in ground water systems, was used to compare the effects of 11 remediation scenarios on a benzene plume. The plume was created in three different hydrogeologic settings from the simulation of an underground gasoline storage tank leak.
Concentration-dependent Effects of D-Methylphenidate on Frontal Cortex and Spinal Cord Networks in vitro
Spontaneously active frontal cortex and spinal cord networks grown on microelectrode arrays were used to study effects of D-methylphenidate. These central nervous system tissues have relatively low concentrations of dopaminergic and noradrenergic neurons compared to the richly populated loci, yet exhibit similar neurophysiological responses to methylphenidate. The spontaneous spike activity of both tissues was inhibited in a concentration-dependent manner by serial additions of 1-500 µM methylphenidate. Methylphenidate is non-toxic as spike inhibition was recovered following washes. The average concentrations for 50% spike rate inhibition (IC50 ± SD) were 118 ± 52 (n= 6) and 57 ± 43 (n = 11) for frontal cortex and spinal cord networks, respectively. A 3 hour exposure of a network to 1 mM methylphenidate was nontoxic. The effective concentrations described in this study are within the therapeutic dosage range. Therefore, the platform may be used for further investigations of drug mechanisms.