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DNA Typing of HLA-B by PCR with Primer Mixes Utilizing Sequence-Specific Primers

Description: The aim of this study was to design a resolution typing system for the HLA-B gene. This technique involves a one-step PCR reaction utilizing genomic DNA and sequence-specific primers to determine the specificity of each allele and to produce a larger primer data base ideal for serological analysis. The application of this technique to serological analysis can improve serology detection which is currently hindered by antibody cross-reactivity and the unavailability of useful typing reagents.
Date: August 1997
Creator: Chiu, Angela Chen-Yen

Isolation and analysis of cotton genomic clones encompassing a fatty acid desaturase (FAD2) gene

Description: Polyunsaturated fatty acids are major structural components of plant chloroplast and endoplasmic reticulum membranes. Two fatty acid desaturases (designated FAD2 and FAD3) desaturate 75% of the fatty acids in the endoplasmic reticulum. The w -6 fatty acid desaturase (FAD2) may be responsible for cold acclimation response, since polyunsaturated phospholipids are important in helping maintain plant viability at lowered temperatures. To study regulation of FAD2 gene expression in cotton, a FAD2 gene was isolated from two genomic libraries using an Arabidopsis FAD2 hybridization probe and a cotton FAD2 5¢ -flanking region gene-specific probe, respectively. A cotton FAD2 gene was found to be in two overlapping genomic clones by physical mapping and DNA sequencing. The cloned DNA fragments are identical in size to cotton FAD2 genomic DNA fragments shown by genomic blot hybridization. The cotton FAD2 coding region has 1,155 bp with no introns and would encode a putative polypeptide of 384 amino acids. The cotton FAD2 enzyme has a high identity of 75% with other plant FAD2 enzymes. The enzyme has three histidine-rich motifs that are conserved in all plant membrane desaturases. These histidine boxes may be the iron-binding domains for reduction of oxygen during desaturation. To confirm that this FAD2 enzyme is functional, a plasmid construct containing the cotton FAD2 coding region was transformed into Saccharomyces cerevisiae. The transformed yeast cells were able to catalyze the conversion of oleic acid (C18:1) into linoleic acid (C18:2). The FAD2 gene contains an intron of 2,967 bp in its 5¢ -flanking region, 11 bp upstream from the initiation codon. The intron could be essential for transcriptional regulation of FAD2 gene expression. Several putative promoter elements occur in the 5¢ -flanking region of this gene. A potential TATA basal promoter element occurs at 41 bp upstream from the cap site. Two presumptive helix-loop-helix (bHLH) ...
Date: May 2001
Creator: Kongcharoensuntorn, Wisatre

Nicotinic Acetylcholine Receptor α3 mRNA in Rat Visual System After Monocular Deprivation

Description: In situ hybridization was used to examine effects of monocular enucleation on nicotinic acetylcholine receptor subunit cc3 mRNA in the rat dLGNand visual cortex. After 28 days postoperative, there were no significant differences in α3 mRNA density between the contralateral (deprived) and ipsilateral (non-deprived) sides. The lack of obvious effects of visual deprivation on α3 mRNA density suggests that other factors, possibly intrinsic to dLGNand visual cortex, govern the postnatal expression of α3 mRNA.
Date: August 1997
Creator: Taylor, James H. (James Harvey), 1970-

Unique applications of cultured neuronal networks in pharmacology, toxicology, and basic neuroscience

Description: This dissertation research explored the capabilities of neuronal networks grown on substrate integrated microelectrode arrays in vitro with emphasis on utilizing such preparations in three specific application domains: pharmacology and drug development, biosensors and neurotoxicology, and the study of burst and synaptic mechanisms. Chapter 1 details the testing of seven novel AChE inhibitors, demonstrating that neuronal networks rapidly detect small molecular differences in closely related compounds, and reveal information about their probable physiological effects that are not attainable through biochemical characterization alone. Chapter 2 shows how neuronal networks may be used to classify and characterize an unknown compound. The compound, trimethylol propane phosphate (TMPP) elicited changes in network activity that resembled those induced by bicuculline, a known epileptogenic. Further work determined that TMPP produces its effects on network activity through a competitive inhibition of the GABAA receptor. This demonstrates that neuronal networks can provide rapid, reliable warning of the presence of toxic substances, and from the manner in which the spontaneous activity changes provide information on the class of compound present and its potential physiological effects. Additional simple pharmacological tests can provide valuable information on primary mechanisms involved in the altered neuronal network responses. Chapter 3 explores the effects produced by a radical simplification of synaptic driving forces. With all synaptic interactions pharmacologically limited to those mediated through the NMDA synapse, spinal cord networks exhibited an extremely regular burst oscillation characterized by a period of 2.9 ± 0.3 s, with mean coefficients of variation of 3.7, 4.7, and 4.9 % for burst rate, burst duration, and inter-burst interval, respectively (16 separate cultures). The reliability of expression of this oscillation suggests that it may represent a fundamental mechanism of importance during periods of NMDA receptor dominated activity, such as embryonic and early postnatal development. NMDA synapse mediated activity produces a precise ...
Date: May 2001
Creator: Keefer, Edward W.

Regulation of Escherichia coli pyrBI Gene Expression in Pseudomonas fluorescens

Description: Pseudomonas fluorescens does not appear to regulate the enzymes of de novo pyrimidine biosynthesis at the level of gene expression. Little or no apparent repression of pyr gene expression is observed upon addition of exogenous pyrimidines to the growth medium. The Escherichia coli pyrBI genes for aspartate transcarbamoylase (ATCase) were sized down and cloned into the broad host range plasmid, pKT230. Upon introduction into a P.fluorescenspyrB mutant strain, ATCase showed repression in response to exogenously fed pyrimidine compounds. Thus, it was possible to bring about changes in pyrimidine nucleotide pool levels and in transcriptional regulation of gene expression at the same time.
Date: May 1995
Creator: Shen, Weiping

Age-Dependent Effects Of Chronic GABAA Receptor Blockade In Barrel Cortex

Description: GABAA receptor binding is transiently increased in rat whisker barrels during the second postnatal week, at a time when neurons in the developing rat cortex are vulnerable to excitotoxic effects. To test whether these GABAA receptors might serve to protect neurons from excessive excitatory input, polymer implants containing the GABAA receptor antagonist bicuculline were placed over barrel cortex for a 4-day period in young (postnatal days 8 - 12) and adult rats. In the cortex of young, but not adult rats, the chronic blockade of GABAA receptors resulted in substantial tissue loss and neuron loss. The greater loss of neurons in young rats supports the hypothesis that a high density of GABAA receptors protects neurons from excessive excitatory input during a sensitive period in development.
Date: May 2001
Creator: Gargan, Lynn

Automated Low-cost Instrument for Measuring Total Column Ozone

Description: Networks of ground-based and satellite borne instruments to measure ultraviolet (UV) sunlight and total column ozone have greatly contributed to an understanding of increased amounts of UV reaching the surface of the Earth caused by stratospheric ozone depletion. Increased UV radiation has important potential effects on human health, and agricultural and ecological systems. Observations from these networks make it possible to monitor total ozone decreases and to predict ozone recovery trends due to global efforts to curb the use of products releasing chemicals harmful to the ozone layer. Thus, continued and expanded global monitoring of ozone and UV is needed. However, existing automatic stratospheric ozone monitors are complex and expensive instruments. The main objective of this research was the development of a low-cost fully automated total column ozone monitoring instrument which, because of its affordability, will increase the number of instruments available for ground-based observations. The new instrument is based on a high-resolution fiber optic spectrometer, coupled with fiber optics that are precisely aimed by a pan and tilt positioning mechanism and with controlling programs written in commonly available software platforms which run on a personal computer. This project makes use of novel low-cost fiber optic spectrometer technology. A cost advantage is gained over available units by placing one end of the fiber outdoors to collect sunlight and convey it indoors, thereby allowing the spectrometer and computer to be placed in a controlled environment. This reduces the cost of weatherproofing and thermal compensation. Cost savings also result from a simplified sun targeting system, because only a small pan and tilt device is required to aim the lightweight fiber optic ends. Precision sun-targeting algorithms, optical filter selection, and software to derive ozone from spectral measurements by the spectrometer are a major contribution of this project. This system is a flexible platform ...
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Date: May 2006
Creator: Nebgen, Gilbert Bernard

Neuronal Network Analyses in vitro of Acute Individual and Combined Responses to Fluoxetine and Ethanol

Description: Embryonic murine neuronal networks cultured on microelectrode arrays were used to quantify acute electrophysiological effects of fluoxetine and ethanol. Spontaneously active frontal cortex cultures showed highly repeatable, dose-dependent sensitivities to both compounds. Cultures began to respond to fluoxetine at 3 µM and were shut off at 10-16 µM. EC50s mean ± S.D. for spike and burst rates were 4.1 ± 1.5 µM and 4.5 ± 1.1 µM (n=14). The fluoxetine inhibition was reversible and without effect on action potential wave shapes. Ethanol showed initial inhibition at 20 mM, with spike and burst rate EC50s at 52.0 ± 17.4 mM and 56.0 ± 17.0 mM (n=15). Ethanol concentrations above 100 -140 mM led to cessation of activity. Although ethanol did not change the shape and amplitude of action potentials, unit specific effects were found. The combined application of ethanol and fluoxetine was additive. Ethanol did not potentiate the effect of fluoxetine.
Date: August 2002
Creator: Xia,Yun

Hepatotoxicity of Mercury to Fish

Description: Tissue samples from spotted gar (Lepisosteus oculatus) and largemouth bass (Micropterus salmoides) were collected from Caddo Lake. Gar and bass livers were subjected to histological investigation and color analysis. Liver color (as abs at 400 nm) was significantly correlated with total mercury in the liver (r2 = 0.57, p = 0.02) and muscle (r2 = 0.58, p = 0.01) of gar. Evidence of liver damage as lipofuscin and discoloration was found in both species but only correlated with liver mercury concentration in spotted gar. Inorganic mercury was the predominant form in gar livers. In order to determine the role of mercury speciation in fish liver damage, a laboratory feeding study was employed. Zebrafish (Danio rerio) were fed either a control (0.12 ± 0.002 µg Hg.g-1 dry wt), inorganic mercury (5.03 ± 0.309 µg Hg.g-1 dry wt), or methylmercury (4.11 ± 0.146 µg Hg.g-1 dry wt) diet. After 78 days of feeding, total mercury was highest in the carcass of zebrafish fed methylmercury (12.49 ± 0.369 µg Hg.g-1 dry wt), intermediate in those fed inorganic mercury (1.09 ± 0.117 µg Hg.g-1 dry wt), and lowest in fish fed the control diet (0.48 ± 0.038 µg Hg.g-1 dry wt). Total mercury was highest in the viscera of methylmercury fed zebrafish (11.6 ± 1.86 µg Hg.g-1 dry wt), intermediate in those fed inorganic diets (4.3 ± 1.08 µg Hg.g-1 dry wt), and lowest in the control fish (below limit of detection). Total mercury was negatively associated with fish length and weight in methylmercury fed fish. Condition factor was not associated with total mercury and might not be the best measure of fitness for these fish. No liver pathologies were observed in zebrafish from any treatment.
Date: August 2010
Creator: Barst, Benjamin Daniel

Functional Neural Toxicity and Endocrine Responses in Mice Following Naphthalene Exposure

Description: Polycyclic aromatic hydrocarbons (PAHs) are a well studied and diverse class of environmental toxicants. PAHs act via the aryl hydrocarbon receptor (AhR), and studies have suggested that PAHs may elicit neurological and estrogenic effects. Doses of PAHs between 50 to 150 ppm may elicit neurotoxicity in rodent models. The present study investigated the effects of naphthalene on in vivo steroidogenesis in Swiss Webster male mice, and in vitro neural function of Balb-C/ICR mice frontal cortex neurons. These data suggest that naphthalene may not elicit steroidogenic effects at concentrations ranging from 0.2 to 25 mg/kg/day, following a 7 day subcutaneous dosing regime. In addition, naphthalene may cause functional toxicity of frontal cortex neurons at concentrations of 32 to 160 ppm naphthalene.
Date: August 2010
Creator: Colbert, Crystal

Mechanisms of rapid receptive field reorganization in rat spinal cord

Description: Rapid receptive field (RF) reorganization of somatosensory neurons in the rat dorsal horn was examined using extracellular single unit recording. Subcutaneous injection of lidocaine into RFs of dorsal horn neurons results in expansion of their RFs within minutes. The expanded RFs appear adjacent to or/and proximal to original RFs. Out of 63 neurons tested, 36 (58%) show RF reorganization. The data suggest that dorsal horn of spinal cord is one of the initial sites for RF reorganization. The neural mechanisms of this effect are not well understood. We propose that changes in biophysical properties (membrane conductance, length constant) of the neurons resulting from lidocaine injection contribute to RF reorganization. Iontophoretic application of glutamate onto dorsal horn neurons that show lidocaine induced RF's expansion were used to test the model. Application of glutamate produced reduction of reorganized RFs in 9 of 20 (45%) tested cells. Application of NBQX produced no effect on either original or expanded RFs indicate that RF shrinkage effects of glutamate involve NMDA receptors. The results are consistent with the prediction of the proposed model. Subcutaneous injection of capsaicin into tactile RFs of low threshold mechanoreceptive dorsal horn neurons produced no effect on the RF sizes that are consistent with other studies. Following the injection, the original RFs were completely silenced (46%) or remained responsive (54%).
Date: August 2002
Creator: Vu, Hung

Water Quality Aspects of an Intermittent Stream and Backwaters in an Urban North Texas Watershed

Description: Pecan Creek flows southeast through the City of Denton, Texas. Characterized as an urban watershed, the basin covers approximately 63.5 km2. Pecan Creek is an intermittent stream that receives nonpoint runoff from urban landuses, and the City of Denton's wastewater treatment plant, Pecan Creek Water Reclamation Plant, discharges effluent to the stream. Downstream from the City of Denton and the wastewater treatment plant, Pecan Creek flows about 6,000 m through agricultural, pasture, and forested landscapes into Copas Cove of Lake Lewisville, creating backwater conditions. Pecan Creek water quality and chemistry were monitored from August 1997 to October 2001. Water quality was influenced by seasonal, spatial, climatic, and diurnal dynamics. Wastewater effluent discharged from the Pecan Creek Water Reclamation Plant had the greatest influence on water quality of the stream and backwaters. Water quality monitoring of Pecan Creek demonstrated that dissolved oxygen standards for the protection of aquatic life were being achieved. Water quality modeling of Pecan Creek was completed to assess future increases in effluent flow from the Pecan Creek Water Reclamation Plant. Water quality modeling indicated that dissolved oxygen standards would not be achieved at the future effluent flow of 21 MGD and at NPDES permitted loadings. Model results with application of a safety factor indicated that the maximum allowable concentrations for a 21 MGD discharge would be 2.3 mg/L of ammonia and 7.0 mg/L of biochemical oxygen demand at summer conditions. Drought conditions that occurred from 1998 to 2001 reduced water levels in Lake Lewisville and impacted dissolved oxygen water quality in Pecan Creek. Water quality observations made during the period of drought allowed for the development of a model to estimate the zone of the dissolved oxygen sag in Pecan Creek based on reservoir elevation. Finally, monitoring results were analyzed with nonparametric statistical procedures to detect water ...
Date: August 2002
Creator: Taylor, Ritchie Don

Use of GIS and Remote Sensing Technologies to Study Habitat Requirements of Ocelots, Leopardus pardalis, in south Texas

Description: The goals of this study were to use Geographic Information Systems (GIS) and remote sensing technologies to gain a better understanding of habitat requirements of a population of ocelots in south Texas, and then apply this knowledge to form a predictive model to locate areas of suitable habitat in Willacy and Cameron counties, Texas. Satellite imagery from August 1991 and August 2000 were classified into four land cover types: closed canopy, open canopy, water, and urban/barren. These classified images were converted into digital thematic maps for use in resource utilization studies and modeling. Location estimates (762 from 1991 and 406 from 2000) were entered into a GIS in order to extract information about home range and resource selection. Each animal's home range was calculated using both Minimum Convex Polygon (MCP) and Kernel home range estimators (95% and 50%). Habitat parameters of interest were: soil, land cover, human density, road density, and distance to closest road, city and water body. Ocelots were found to prefer closed canopy and avoid open canopy land cover types. Ocelots preferred soils known to support thorn scrub, an indication of the importance of this habitat. Landscape metrics associated with habitat used by ocelots were determined through the use of Patch Analyst, an extension for ArcView 3.2. Contrary to expectations, ocelots utilized areas with greater fragmentation than random areas available for use. However, this use of highly fragmented areas was an indication of the degree of fragmentation of suitable habitat in the area. Further investigation of patch size selection indicated that ocelots used large sized patches disproportionately to availability, indicating a preference for larger patches. A model was created using the resource selection and habitat preference GIS database from 1991. This model was used to identify areas of “optimal”, ”sub-optimal”, and “unsuitable” habitat for ocelots in 2000. ...
Date: August 2002
Creator: Jackson, Victoria L.

Effects of Sublethal Copper Exposure on Escape Behavior and Growth of Rana pipiens Tadpoles

Description: This research is designed to test how sublethal exposure to copper affects tadpole predator-escape behavior and how quickly tadpoles recover. After exposure, tadpoles were separated. Escape behavior was recorded for two-thirds of exposed tadpoles while one-third of the exposed population was measured weekly to determine growth and recovery. Control tadpoles were consumed within 15 minutes whereas those exposed to higher concentrations were consumed at a slower rate, which does not support the hypotheses. Although the rate of predation was lower, tadpoles exposed to higher Cu concentrations were on average, 1.47 cm in total body length. Those exposed to 0.93 mg/L averaged 0.86 cm. After being placed into clean water, treatment tadpoles recovered after 20 days.
Date: May 2002
Creator: Redick, Melinda

Removal of selected water disinfection byproducts, and MTBE in batch and continuous flow systems using alternative sorbents.

Description: A study was conducted to evaluate the sorption characteristics of six disinfection byproducts (DBPs) on four sorbents. To investigate sorption of volatile organic compounds (VOCs), specially designed experimental batch and continuous flow modules were developed. The investigated compounds included: chloroform, 1,2-dichloroethane (DCE), trichloroethylene (TCE), bromodichloromethane (BDCM), methyl tertiary butyl ether (MTBE), bromate and bromide ions. Sorbents used included light weight aggregate (LWA), an inorganic porous material with unique surface characteristics, Amberlite® XAD-16, a weakly basic anion exchange resin, Amberjet®, a strongly basic anion exchange resin, and granular activated carbon (GAC). Batch experiments were conducted on spiked Milli-Q® and lake water matrices. Results indicate considerable sorption of TCE (68.9%), slight sorption of bromate ions (19%) and no appreciable sorption for the other test compounds on LWA. The sorption of TCE increased to 75.3% in experiments utilizing smaller LWA particle size. LWA could be a viable medium for removal of TCE from contaminated surface or groundwater sites. Amberlite® was found unsuitable for use due to its physical characteristics, and its inability to efficiently remove any of the test compounds. Amberjet® showed an excellent ability to remove the inorganic anions (>99%), and BDCM (96.9%) from aqueous solutions but with considerable elevation of pH. Continuous flow experiments evaluated GAC and Amberjet® with spiked Milli-Q® and tap water matrices. The tested organic compounds were sorbed in the order of their hydrophobicity. Slight elevation of pH was observed during continuous flow experiments, making Amberjet® a viable option for removal of BDCM, bromate and bromide ions from water. The continuous flow experiments showed that GAC is an excellent medium for removal of the tested VOCs and bromate ion. Each of the test compounds showed different breakthrough and saturation points. The unique design of the continuous flow apparatus used in the study proved to be highly beneficial to ...
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Date: December 2002
Creator: Kadry, Ahmed Y.

Use of geographic information systems for assessing ground water pollution potential by pesticides in central Thailand

Description: This study employed geographic information systems (GIS) technology to evaluate the vulnerability of groundwater to pesticide pollution. The study area included three provinces (namely, Kanchana Buri, Ratcha Buri, and Suphan Buri) located in the western part of central Thailand. Factors used for this purpose were soil texture, percent slope, primary land use, well depth, and monthly variance of rainfall. These factors were reclassified to a common scale showing potential to cause groundwater contamination by pesticides. This scale ranged from 5 to 1 which means high to low pollution potential. Also, each factor was assigned a weight indicating its influence on the movement of pesticides to groundwater. Well depth, the most important factor in this study, had the highest weight of 0.60 while each of the remaining factors had an equal weight of 0.10. These factors were superimposed by a method called “arithmetic overlay” to yield a composite vulnerability map of the study area. Maps showing relative vulnerability of groundwater to contamination by pesticides were produced. Each of them represented the degree of susceptibility of groundwater to be polluted by the following pesticides: 2,4-D, atrazine, carbofuran, dicofol, endosulfan, dieldrin & aldrin, endrin, heptachlor & heptachlor epoxide, total BHC, and total DDT. These maps were compared to groundwater quality data derived from actual observations. However, only the vulnerability maps of atrazine, endosulfan, total BHC, and heptachlor & heptachlor epoxide showed the best approximation to actual data. It was found that about 7 to 8%, 83 to 88% and 4.9 to 8.7% of the study area were highly, moderately, and lowly susceptible to pesticide pollution in groundwater, respectively. In this study a vulnerability model was developed, which is expressed as follow: V = 0.60CW + 0.10CS + 0.10CR + 0.10CL + 0.10CSL. Its function is to calculate a vulnerability score for a certain ...
Date: August 2002
Creator: Thapinta, Anat

Influence of Acclimation and Acclimatization to Seasonal Temperatures on Metabolism and Energetics in the Rusty Lizard Sceloporus Olivaceus

Description: Rates and energy equivalents of consumption (C), egestion+ excretion (FU) , assimilation (A), respiration (R) and production (P) were measured in two groups of Sceloporus olivaceus: 1) a laboratory group acclimated at four seasonally encountered temperatures (15, 20, 25 and 30°C) ; and 2) four acclimatization groups collected at different seasons when ambient temperatures corresponded to 15, 20, 25 and 30°C.
Date: May 1973
Creator: Dutton, Ronald Hugh

Experimental Trichinosis in Birds

Description: This work concerns itself with essentially four experiments: (1) the cecum-injective-infection experiment; (2) the anus-injective-infection experiment; (3) the mouth ingestive-infection with larvae, and (4) the mouth ingestive-infection with the flesh of infected rats.
Date: December 1970
Creator: Poon, Yau-Lun

Mixed Culture of Chlorella Pyrenoidosa TX71105 and a Variant Strain of Bacillus Megaterium

Description: Very little work has been done on bacteria capable of significantly inhibiting algal growth. This thesis reports the research on mixed cultures of a high-temperature strain of algae, Chlorella pyrenoidosa TX71105, and an organism isolated from the air and tentatively identified as a variant strain of Bacillus megaterium.
Date: August 1970
Creator: Yao, Raymond Che-Fong

Studies on the Morphology and Biology of Cotton Rats (Sigmodon hispidus) from Northern Mexico to Southern Nebraska

Description: This investigation was designed to evaluate the need for retaining both Sigmodon hispidus texianus and Sigmodon hispidus berlandieri as subspecific designations. An attempt was made to demonstrate bioclimatic variation and reproductive seasonality in cotton rats. The validity of applying the results of isolated studies of cotton rat populations to the species as a whole was examined.
Date: December 1971
Creator: Cleveland, Arthur Gordon

Some Acute Effects of X-Irradiation (LD100) on Plasma and Adrenal Tissue Histamine in Rats

Description: The effects of a lethal dose (1380 r) of X-irradiation on plasma and adrenal tissue histamine levels of rats were studied. The plasma histamine response was triphasic (increase at 1-3 hours, decrease at 5 and 9 hours and return to control at 24 hours post-irradiation). The adrenal tissue histamine response was found to be biphasic (decrease at 1 to 9 hours and a return to control level at 24 hours post-irradiation).
Date: May 1972
Creator: Ferguson, James L.

Obligately Thermophilic Nitrogen-Fixation in Some Soil Bacteria

Description: In the work presented here, it is claimed that bacteria have been isolated which are capable of growth at high temperatures utilizing molecular nitrogen as their sole nitrogen source. Soil bacteria were isolated which grew at 55 C in nitrogen-free media. They were found to be obligatory thermophiles in nitrogen-free media and facultative thermophiles in media containing organically bound nitrogen.
Date: August 1971
Creator: Milam, Mary