The human opportunistic pathogen, Pseudomonas aeruginosa PAO1, has been shown to kill the nematode Caenorhabditis elegans. C. elegans has been a valuable model for the study of bacterial pathogenesis, and has reinforced the notion that common virulence and host defense mechanisms exist. Recently, the pyrimidine pathway was shown to regulate virulence levels. Therefore, mutations in the pyrimidine pathway of PAO1 showed decrease virulence in the nematode. When starving the nematode, bacterial resistance was also shown to increase. It was hypothesized that starvation induced the DAF pathway, which regulates the transcription of genes involved with the antibacterial defense mechanism. Further research will be conducted to test this theory by performing RNAi experiments for the genes functioning in the antibacterial defense mechanism.

To examine if a rhythm can be entrained in either heart rate or oxygen consumption in late stage embryos (days 17-19.5) with light as a zeitgeber, chicken embryos were incubated in complete darkness (D:D) and 12:12 light:dark cycle (L:D). Light had no impact on oxygen consumption (390 µL O2∙min-1∙egg-1) but increased heart rate for non-internally pipped embryos (260 to 270 beats∙min-1 during light cycle). Oxygen consumption increased independent of pipping while heart rate increased (255 to 265 beats∙min-1) in D:D embryos due to pipping. A light-induced rhythm or effect occurred in heart rate but not oxygen consumption, suggesting heart rate and oxygen consumption may be uncoupled.

White-tailed deer (Odocoileus virginianus) (N=15) movements were determined with use of radio telemetry techniques to determine the utilization of corridor habitat on the Lake Ray Roberts Greenbelt Corridor (RRGC) in north central Texas. Home ranges were calculated using three estimation types. Male white-tailed deer tend to have home ranges twice that of female home ranges. Seasonal home ranges were largest during spring (Feb. - April) and fall (Aug. - Oct.) seasons. Males had greater seasonal variation in utilization than females. No statistically significant difference (p=0.24) between white-tailed deer locations when the RRGC experiences heavy human traffic compared to days when there is light human traffic. Linearity indices indicated home ranges less linear than expected (LI = 3.02). The RRGC should be maintained at its current status to provide a variety of vegetational types and protective cover for white-tailed deer and other wildlife of Denton County.

The Consumer's Guide is designed to help consumers by providing guidelines for the purchase of specific energy-efficient household appliances- water heaters, air conditioning and heating systems, windows, dishwashers, refrigerators, clothes washers, and dryers. This serves two major purposes: to decrease the environmental impact of those products and to save consumers money over the lifetime of the products. The seven major appliances covered in this work are things that consumers tend to purchase quickly when their older models wear out and with little research into their energy and/or water efficiency. The guide begins with a general introduction and an explanation of the need for energy conservation. Explanations of how they work, purchasing tips, installation tips, maintenance tips, tips for additional energy efficiency, and case studies are given for each appliance. Printable pamphlets are included at the end.

The focus of this study was to characterize the surface water and groundwater hydrology of borrow-pit wetlands located within the borders of the Lewisville Lake Environmental Learning Area (LLELA), east of the Elm Fork of the Trinity River. The wetlands were excavated into alluvial deposits downstream of the Lewisville Lake Dam. Both surface water and groundwater contribute to the hydro-period of the borrow-pit wetlands. Nearby marshes exhibit characteristics of groundwater discharge. Salinity in groundwater-fed wetlands could affect establishment of vegetation, as suggested from plant surveys. Surface water input from storm events dilutes salinity levels. Management of LLELA wetlands should include long-term evaluation of hydrology and plantings to enhance habitat. Plans for additional wetlands should consider both surface water and groundwater inputs.

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 …

Several key flexure sites exist in the muscle crossbridge including the actomyosin binding site which play important roles in the actomyosin crossbridge cycle. To distinguish between these sources of flexibility, a new single molecule assay was developed to observe the swiveling of rod about a single myosin. Myosins attached through a single crossbridge displayed mostly similar torsional characteristics compared to myosins attached through two crossbridges, which indicates that most of the torsional flexibility resides in the myosin subfragment-2, and thus the hinge between subfragment-2 and light meromyosin should contribute the most to this flexibility. The comparison of torsional characteristics in the absence and presence of ADP demonstrated a small but significant increase in twist rates for the double-headed myosins but no increase for single-headed myosins, which indicates that the ADP-induced increase in flexibility arises due to changes in the myosin head and verifies that most flexibility resides in myosin subfragment-2.

The pyrimidine biosynthetic pathway is an essential pathway for most organisms. Previous research on the pyrimidine pathway in Pseudomonas aeruginosa (PAO1) has shown that a block in the third step of the pathway resulted in both a requirement for exogenous pyrimidines and decreased ability to produce virulence factors. In this work an organism with a mutation in the second step of the pathway, aspartate transcarbamoylase (ATCase), was created. Assays for pyrimidine intermediates, and virulence factors were performed. Results showed that the production of pigments, haemolysin, and rhamnolipids were significantly decreased from PAO1. Elastase and casein protease production were also moderately decreased. In the Caenorhabditis elegans infection model the nematodes fed the ATCase mutant had increased mortality, as compared to nematodes fed wild type bacteria. These findings lend support to the hypothesis that changes in the pyrimidine biosynthetic pathway contribute to the organism's ability to effect pathogenicity.

Histological methods to estimate skeletal age at death, in forensic cases, are an alternative to the more traditional gross morphological methods. Most histological methods utilize counts of bone type within a given field for their estimation. The method presented in this paper uses the percentage area occupied by unremodeled bone to estimate age. The percentage area occupied by unremodeled bone is used in a linear regression model to predict skeletal age at death. Additionally, this method uses digital software to measure area rather than the traditional technique in which a gridded microscope is used to estimate area. The clavicle was chosen as a sample site since it is not a weight bearing bone and has little muscular insertion. These factors reduce the variation seen as a result of differences in lifestyle or activity pattern.

The primary focus of the study was to compare dissolved organic carbon (DOC) at baseline stream flow to DOC at a higher post-rain stream flow, with a secondary focus on comparison of DOC between sites. Comparisons were also done on suspended solids at baseline flow to those of the higher post-rain flow, as well as suspended solids between sites. Significant differences did exist between DOC sampled at baseline flow and DOC in samples taken at peak flow. The study found no difference in suspended solids among sites neither on either baseflow sampling nor on the post rain event sampling.

A new subspecies of the plains pocket gopher (Geomys bursarius ozarkensis), located in the Ozark Mountains of north central Arkansas, was recently described by Elrod et al. (2000). Current range for G. b. ozarkensis was established, habitat preference was assessed by analyzing soil samples, vegetation and distance to stream and potential pocket gopher habitat within the current range was identified. A census technique was used to estimate a total density of 3, 564 pocket gophers. Through automobile and aerial survey 51 known fields of inhabitance were located extending the range slightly. Soil analyses indicated loamy sand as the most common texture with a slightly acidic pH and a broad range of values for other measured soil parameters and 21 families of vegetation were identified. All inhabited fields were located within an average of 107.2m from waterways and over 1,600 hectares of possible suitable habitat was identified.

Hematocrit and hematocrit regulation have the potential to affect developing embryos. To examine the ability of chicken embryos at day 15 to regulate hematocrit, they were subjected to either repeated saline injections (5% of total blood volume) or repeated blood removal (5% of total blood volume). Embryos showed an ability to maintain hematocrit (~20%) despite blood volume increases up to 115% of initial blood volume. Embryos were not able to maintain hematocrit in the face of dramatic blood volume loss. Oxygen consumption of embryos could be affected by their level of hematocrit. To examine this, chicken embryos at day 15, 16, and 17 of incubation were given a high hematocrit (~50-60%) sample of blood (400 μl) to artificially increase the hematocrit of the embryos (~10-12%). Despite the increase in oxygen availability, when monitored over a period of six hours, embryos showed no difference (0.36 ± 0.01 (ml O2 - min-1- egg-1) in metabolism from baseline measurements at day 15, 16 and 17.

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.

Robust Vallisneria americana was cultured for restoration purposes. Preliminary studies, with various iron treatments, were conducted to ascertain the amount of phosphorous release into the water column from sediments. There was a significant difference in the amount of phosphorous released if commercial sediment was used with a low iron amendment or without an iron amendment. The second study consisted of planting V. americana on two different sediment types while supplying half of the plants with additional CO2. Plants grown on pond sediment with additional CO2 had significantly more biomass. In the third study all plants were grown on pond sediment, and half were treated with CO2. All plants that were treated with additional CO2 had significantly more biomass than those that were aerated.

TNT (2,4,6-trinitrotoluene) readily and predominantly transforms to 2ADNT (2-amino-4,6-dinitrotoluene) and 4ADNT (4-amino-2,6-dinitrotoluene) in environmental matrixes and tissues. Solid phase microextraction (SPME) was used to extract ADNTs (amino-dinitrotoluenes) from tissue as a potential method to investigate the recalcitrance of metabolically-generated ADNTs versus absorbed ADNTs. Tubifex tubifex was allowed to metabolize TNT into ADNTs in 24-hr static non-renewal exposure test followed by 24-hr depuration in clean reconstituted hard water. Polyacrylate-coated (PA) SPME fibers were then deployed and agitated in tissue homogenates containing metabolically-generated ADNTs for 48 hr to provide a measure of available ADNTs. Extractability of ADNTs from T. tubifex tissue containing metabolically-generated ADNTs was significantly less than extractability of ADNTs from T. tubifex tissue containing absorbed ADNTs: 50-60% and 81-90% of expected extractability based on fiber-water partition ratio. The lower SPME extractability of metabolically-generated ADNTs may stem from the unavailability of metabolically-generated ADNTs sequestered in tissue or bound to tissue macromolecules during metabolism of TNT to ADNT. Tissue extractions using SPMEs may be able to estimate such bound organic residues in tissue and serve as potential indicators of toxicological bioavailability and biomagnification potential of tissue-associated organic compounds.

A hydrologic model and watershed export model was used to estimate the loading of nutrients and total suspended solids from un-gaged local watersheds associated with Lake Texoma. Discharge to the reservoir from local watersheds was predicted using a modification of the curve number method in HEC Hydrologic Modeling System (HEC-HMS). Event mean concentrations were applied to land use to estimate loadings of nutrients and TSS. Total discharge from the local watersheds was estimated to be 3.02 x 107 cubic meters for a study window of March 1 to May 31, 1997, less than 10 percent of the input to the lake from the Red River and Washita River systems. Loadings were estimated to be 33,553 kg nitrogen, 4,401 kg phosphorus, and 3,423,140 kg TSS. The models and results obtained from their application appear to have potential utility for use in a water quality management decision support system for the reservoir.

The purpose of this research was to study the hybridization of molecular beacons under different conditions and designs. Data collected suggest that the inconsistency found in the emission intensity of several of these probes may be caused by 3 important factors: length of the probe, nucleotide sequence and, the formation of an alternative complex structure such as a dimer. Of all three factors, dimer formation is the most troublesome, since it reduces the emission of the reporter molecules. A new probe design was used to reduce dimer formation. The emission signal of the improved probe was several folds stronger than those probes with the early design. In this research, dimer formation is detected, furthermore a new probe with a different design was tested. If dimer formation can be reduced molecular beacons can be integrated into more complex hybridization systems providing an important tool in research and diagnosis of genetic disorders.

The Pecan Creek Waste Reclamation Plant in Denton, Texas, an activated sludge WWTP, was sampled monthly for ten months to determine seasonal and site variation in concentrations of triclosan (5-chloro-2-(2,4-dichlorophenoxy)phenol), an antibacterial additive. SNK separation after the highly significant ANOVA on ranked data were: summer = fall > winter = spring and influent > downstream = effluent = wetland inflow > wetland outflow (a=0.05). After the plant converted to ultraviolet disinfection, measurements were made before and after the UV basin to determine if significant amounts of triclosan were converted to dioxin. Percent loss at each of the treatment steps was determined. Concentrations of triclosan in the downstream site were below the published NOEC for the most sensitive species.

The lipid composition of cotton (Gossypium hirsutum, L) fibers was determined. Fatty acid profiles revealed that linolenate and palmitate were the most abundant fatty acids present in fiber cells. Phosphatidylcholine was the predominant lipid class in fiber cells, while phosphatidylethanolamine, phosphatidylinositol and digalactosyldiacylglycerol were also prevalent. An unusually high amount of phosphatidic acid was observed in frozen cotton fibers. Phospholipase D activity assays revealed that this enzyme readily hydrolyzed radioactive phosphatidylcholine into phosphatidic acid. A profile of expressed sequence tags (ESTs) for genes involved in lipid metabolism in cotton fibers was also obtained. This EST profile along with our lipid metabolite data was used to predict lipid metabolic pathways in cotton fiber cells.