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The Role of Thyroid Hormone across Avian Development Spectrum: Investigations on Systemic Development, Metabolism and Ontogeny of Endothermy

Description: Achievement of endothernic capacity is vital for independence from ambient temperature changes, sustained activity, optimal biochemical reactions and optimization of parental care. During early avian development, the core tenets of transition from ectothermy to endothermy are development of metabolic capacity (oxygen consumption, mitochondrial bioenergetics), enhanced cardiovascular function (heart rate and cardiac output), pulmonary ventilation and thermogenic capacity. Thyroid hormones, particularly T3, are key metabolic regulators of basal metabolism, thermogenesis, pulmonary ventilation and mitochondrial respiration. Thyroid hormone fluctuation patterns during both precocial and altricial avian endothermic transition suggest a prominent role in maturation of endothermy, cardiovascular, respiratory and skeletal muscle physiology. This body of work explores effects of T3 manipulations in two avian species: the precocial Pekin duck and the altricial Red-winged Blackbird. Increased plasma T3 during late incubation resulted in increased cardiac mass, elevated resting and intrinsic heart rate, intrinsic mean arterial pressure, increased cholinergic tone and blunted alpha-adrenergic tone in the precocial Pekin duck. In both Pekin duck and Red-winged blackbird, plasma T3 levels correlated with changes in the trajectory of endothermic ontogeny, systemic oxygen consumption, thermogenesis, maturation of pulmonary ventilatory function, altered growth and effects on skeletal and cardiac mitochondrial bioenergetics. These observations support the role of thyroid hormones as metabolic and developmental regulators at the time of attainment of endothermy during the perinatal period in precocial and altricial avian species. Insights into the role of thyroid hormone as a metabolic and development regulator at the time of avian endothermic attainment provide a more thorough understanding of metabolic and physical transitions a hatchling bird must undergo to reach the adult endothermic phenotype. Such insights also deepen understanding of the complex role thyroid hormones play in homeostasis and offer implications about the evolutionary history of endothermic capacity.
Date: August 2017
Creator: Sirsat, Tushar S

Delivery of CRISPR/Cas9 into Blood Cells of Zebrafish: Potential for Genome Editing in Somatic Cells

Description: Factor VIII is a clotting factor found on the intrinsic side of the coagulation cascade. A mutation in the factor VIII gene causes the disease Hemophilia A, for which there is no cure. The most common treatment is administration of recombinant factor VIII. However, this can cause an immune response that renders the treatment ineffective in certain hemophilia patients. For this reason a new treatment, or cure, needs to be developed. Gene editing is one solution to correcting the factor VIII mutation. CRISPR/Cas9 mediated gene editing introduces a double stranded break in the genomic DNA. Where this break occurs repair mechanisms cause insertions and deletions, or if a template oligonucleotide can be provided point mutations could be introduced or corrected. However, to accomplish this goal for editing factor VIII mutations, a way to deliver the components of CRISPR/Cas9 into somatic cells is needed. In this study, I confirmed that the CRISPR/Cas9 system was able to create a mutation in the factor VIII gene in zebrafish. I also showed that the components of CRISPR/Cas9 could be piggybacked by vivo morpholino into a variety of blood cells. This study also confirmed that the vivo morpholino did not interfere with the gRNA binding to the DNA, or Cas9 protein inducing the double stranded break.
Date: August 2017
Creator: Schneider, Sara Jane

Markov Model of Segmentation and Clustering: Applications in Deciphering Genomes and Metagenomes

Description: Rapidly accumulating genomic data as a result of high-throughput sequencing has necessitated development of efficient computational methods to decode the biological information underlying these data. DNA composition varies across structurally or functionally different regions of a genome as well as those of distinct evolutionary origins. We adapted an integrative framework that combines a top-down, recursive segmentation algorithm with a bottom-up, agglomerative clustering algorithm to decipher compositionally distinct regions in genomes. The recursive segmentation procedure entails fragmenting a genome into compositionally distinct segments within a statistical hypothesis testing framework. This is followed by an agglomerative clustering procedure to group compositionally similar segments within the same framework. One of our main objectives was to decipher distinctive evolutionary patterns in sex chromosomes via unraveling the underlying compositional heterogeneity. Application of this approach to the human X-chromosome provided novel insights into the stratification of the X chromosome as a consequence of punctuated recombination suppressions between the X and Y from the distal long arm to the distal short arm. Novel "evolutionary strata" were identified particularly in the X conserved region (XCR) that is not amenable to the X-Y comparative analysis due to massive loss of the Y gametologs following recombination cessation. Our compositional based approach could circumvent the limitations of the current methods that depend on X-Y (or Z-W for ZW sex determination system) comparisons by deciphering the stratification even if only the sequence of sex chromosome in the homogametic sex (i.e. X or Z chromosome) is available. These studies were extended to the plant sex chromosomes which are known to have a number of evolutionary strata that formed at the initial stage of their evolution, presenting an opportunity to examine the onset of stratum formation on the sex chromosomes. Further applications included detection of horizontally acquired DNAs in extremophilic eukaryote, Galdieria sulphuraria, which ...
Date: August 2017
Creator: Pandey, Ravi Shanker

Effects of Brain Brain Injury on Primary Cilia of Glial Cells and Pericytes

Description: Glial cells maintain homeostasis that is essential to neuronal function. Injury to the nervous system leads to the activation and proliferation of glial cells and pericytes, which helps to wall off the damaged region and restore homeostatic conditions. Sonic hedgehog is a mitogen which is implicated in injury-induced proliferation of glial cells and pericytes. The mitogenic effects of sonic hedgehog require primary cilia, but the few reports on glial or pericyte primary cilia do not agree about their abundance and did not address effects of injury on these cilia. Primary cilia are microtubule-based organelles that arise from the centrosome and are retracted before cells divide. Depending on cell type, proteins concentrated in cilia can transduce several mitotic, chemosensory, or mechanosensory stimuli. The present study investigated effects of stab wound injury on the incidence and length of glial and pericyte primary cilia in the area adjacent to the injury core. Astrocytes, polydendrocytes and pericytes were classified by immunohistochemistry based on cell-type markers. In normal adult mice, Arl13b immunoreactive primary cilia were present in a majority of each cell type examined: astrocytes, 98±2%; polydendrocytes, 87±6%; and pericytes, 79±13% (mean ± SEM). Three days post-injury, cilium incidence decreased by 24% in astrocytes (p< 0.008) and 41% in polydendrocytes (p< 0.002), but there was no significant effect in pericytes. Polydendrocytes labeled with the cell cycle marker Ki67 were less likely to have cilia compared to resting, Ki67- polydendrocytes. Considering post-injury rates of proliferation for astrocytes and polydendrocytes, it appears that resorption of cilia due to cell cycle entry may account for much of the loss of cilia in polydendrocytes but was not sufficient to account for the loss of cilia in astrocytes. Under normal conditions, astrocytes rarely divide, and they maintain non-overlapping territories. However, three days after injury, there was a 7-fold increase in ...
Date: December 2016
Creator: Coronel, Marco Vinicio

Environmental Modulation of the Onset of Air-breathing of the Siamese Fighting Fish and the Blue Gourami

Description: This study determined the effect of hypoxia on air-breathing onset and physiological and morphological characters in larvae of the air breathing fishes Trichopodus trichopterus and Betta splendens. Larvae were exposed intermittently (12/12 h daily) to 20, 17, and 14 kPa of PO2 from 1 to 40 days post-fertilization. Survival, onset of air breathing, wet body mass, O2, Pcrit were measured every 5 dpf. Hypoxia advanced by 4 days, and delayed by 9 days, the onset of air breathing in Betta and Trichopodus, respectively. Hypoxia increased larval body length, wet mass, and labyrinth organ respiratory surface of Betta, but did not affect these factors in Trichopodus. Hypoxic exposure increased O2 by 50-100% at each day throughout larval development in Betta, but had no effect on larval Trichopodus. Hypoxia decreased Pcrit in Betta by 37%, but increased Pcrit in Trichopodus by 70%. Larval Betta reared in hypoxia showed a modified heart rate:opercular rate ratio (3:1 to 2:1), but these changes did not occur in Trichopodus. Compared to Betta, the blood of Trichopodus had a higher P50 and much smaller Bohr and Root effects. These interspecific differences are likely due to ecophysiological differences: Betta is a non- obligatory air-breather after 36 dpf with a slow lifestyle reflected in its low metabolism, while Trichopodus is an obligatory air-breather past 32 dpf with an athletic fast lifestyle and accompanying high metabolism.
Date: December 2015
Creator: Mendez Sanchez, Jose Fernando

Effect of Acute Alcohol Ingestion on Resistance Exercise Induced mTORC1 Signaling in Human Muscle

Description: The purpose of this project was to further elucidate the effects post-exercise alcohol ingestion. This project had many novel aspects including using a resistance exercise (RE) only exercise design and the inclusion of women. To our knowledge, we are the first to investigate the effect of post-RE alcohol ingestion in women. In the first chapter of this project, information on the prevalence of alcohol use and the importance of skeletal muscle as a dynamic and metabolic tissue was provided. In chapter two, the effects of post-RE alcohol ingestion in men and women are detailed. The major findings of this study was that although RE elicited similar mTORC1 signaling both in men and in women, alcohol ingestion appeared to only attenuate RE-induced phosphorylation of the mTORC1 signaling pathway in men. The third chapter focused on examining the effects of post-RE alcohol ingestion on acute testosterone bioavailability. The primary findings of this study was that alcohol substantially elevated serum total and free testosterone concentrations during recovery from a bout of resistance exercise. The fourth chapter detailed factors that contribute to bone density in men. The major findings of this study was that young adult male long-distance runners who participated in resistance training at least once per week had greater bone mineral density than their non-resistance trained and non-exercise trained peers.
Date: August 2015
Creator: Duplanty, Anthony A.

The Effect of Curcumin Supplementation on Physical and Biological Indices of Delayed Onset Muscle Soreness and Inflammation Following Muscle Injury

Description: In this project, the effects of dietary polyphenols on exercise-induced muscle damage and vascular health are examined. Dietary polyphenols exert well-known anti-inflammatory effects; however, how these effects are realized with respect to vascular health and EIMD is relatively unknown. I begin by reviewing the available literature surrounding the impact of three dietary polyphenols (curcumin, catechins, and quercetin) on inflammation associated with EIMD. It is well established that their primary means of anti-inflammation is through alterations of NF-κB and AP-1 transcription activities. Given this, their inclusion into training strategies seems reasonable. Consistent evidence is presented making a case for the anti-inflammatory effects of dietary polyphenols following EIMD. I follow this review up by completing an in-depth study on the consumption of curcumin prior to EIMD. I found curcumin (1000 mg/day) can reduce subjective soreness and decrease inflammation compared to placebo controls. To further understand the effects of dietary polyphenols on health, I investigate the effects of a four-week supplementation period of cocoa (catechins) on vascular. I concluded that atherogenic risk in obese women is reduced after consumption of cocoa. In addition to these experimental projects, I developed two novel methods that can be used to investigate vascular health (EMP concentration) and intracellular protein and mRNA production using flow cytometry.
Date: May 2015
Creator: Venable, Adam Steven

Understanding Microbial Biodegradation of Environmental Contaminants

Description: The accumulation of industrial contaminants in the natural environments have rapidly become a serious threat for human and animal life. Fortunately, there are microorganisms capable of degrading or transforming environmental contaminants. The present dissertation work aimed to understand the genomic basis of microbial degradation and resistance. The focus was the genomic study of the following bacteria: a) Pseudomonas fluorescens NCIMB 11764, a unique bacterium with specific enzymes that allow cyanide adaptation features. Potential cyanide degradation mechanisms found in this strain included nit1C cluster, and CNO complex. Potential cyanide tolerance genes found included cyanide insensitive oxidases, nitric oxide producing gene, and iron metabolism genes. b) Cupriavidus sp. strain SK-3 and strain SK-4. The genome of both bacteria presented the bph operon for polychlorinated biphenyl (PCB) degradation, but we found differences in the sequences of the genes. Those differences might indicate their preferences for different PCB substrates. c) Arsenic resistant bacterial communities observed in the Atacama Desert. Specific bacteria were found to thrive depending on the arsenic concentration. Examples were Bacteroidetes and Spirochaetes phyla whose proportions increased in the river with high arsenic concentrations. Also, DNA repair and replication metabolic functions seem to be necessary for resistance to arsenic contaminated environments. Our research give us insights on how bacteria communities, not just individually, can adapt and become resistant to the contaminants. The present dissertation work showed specific genes and mechanisms for degradation and resistance of contaminants that could contribute to develop new bioremediation strategies.
Date: May 2015
Creator: Vilo Muñoz, Claudia Andrea

Characteristics of Primary Cilia and Centrosomes in Neuronal and Glial Lineages of the Adult Brain

Description: Primary cilia are sensory organelles that are important for initiating cell division in the brain, especially through sonic hedgehog (Shh) signaling. Several lines of evidence suggest that the mitogenic effect of Shh requires primary cilia. Proliferation initiated by Shh signaling plays key roles in brain development, in neurogenesis in the adult hippocampus, and in the generation of glial cells in response to cortical injury. In spite of the likely involvement of cilia in these events, little is known about their characteristics. Centrosomes, which are associated with primary cilia, also have multiple influences on the cell cycle, and they are important in assembling microtubules for the maintenance of the cell’s cytoskeleton and cilia. The cilia of terminally differentiated neurons have been previously examined with respect to length, incidence, and receptors present. However, almost nothing is known about primary cilia in stem cells, progenitors, or differentiated glial cells. Moreover, it is not known how the properties of cilia and centrosomes may vary with cell cycle or proliferative potential, in brain or other tissues. This dissertation focuses first on neurogenesis in the hippocampal subgranular zone (SGZ). The SGZ is one of the few brain regions in mammals that gives rise to a substantial number of new neurons throughout adulthood. The neuron lineage contains a progression of identifiable precursor cell types with different proliferation rates. This present study found that primary cilia were present in every cell type in the neuronal lineage in SGZ. Cilium length and incidence were positively correlated among these cell types. Ciliary levels of adenylyl cyclase type III (ACIII) levels relative to ADP-ribosylation factor-like protein 13b (Arl13b) was higher in neurons than in precursor cells and glia, and also changed with the cell cycle. G-protein coupled receptors, SstR3, MCHR1, and Gpr161 receptors were only found in neuronal cilia. The levels ...
Date: May 2015
Creator: Bhattarai, Samip Ram

Women Have Higher Skin Temperature on the Back during Treadmill Exercise in a Hot, Humid Environment

Description: A common measurement of body temperature during exercise in a hot, humid environment is mean skin temperature collected from 3-12 sites on the body. However, such an approach fails to demonstrate localized differences in skin temperature that are likely to exist as a function of gender. The purpose of this study was to examine potential differences in skin temperature between men and women at 17 different locations on the body. Young women (21 ± 1 y; n = 11) and men (23 ± 3; n = 10) were recruited to complete a 60-min walk/jog interval protocol in a hot (34 ± 1 °C), humid (64 ± 8%) environment while skin temperature was measured. Data was analyzed using a repeated-measures ANOVA (p < 0.05) and location of interaction effects determined using a Fisher’s least squares difference test. We observed a higher change (p < 0.05) from baseline skin temperatures (ΔTsk) for women in three locations: left upper back (women: avg. ΔTsk = 4.12 ± 0.20 °C; men: avg. ΔTsk = 2.70 ± 0.10 °C), right upper back (women: avg. ΔTsk = 4.19 ± 0.07 °C; men: avg. ΔTsk = 2.92 ± 0.05 °C), and right mid-back (women: avg. ΔTsk = 4.62 ± 0.14 °C; men: avg. ΔTsk =3.55 ± 0.09 °C). Individual time differences between genders occurred after 7- (left upper back) and 15-min (right upper back, right mid-back) of exercise and were maintained until the end of exercise. Women have a greater increase in skin temperature at three locations on the back following the onset of exercise in a hot, humid environment. This report provides important information regarding the implications of women exercising in a hot, humid environment.
Date: August 2014
Creator: Venable, Adam Steven

Acute Effects of the Antibiotic Streptomycin on Neural Network Activity and Pharmacological Responses

Description: The purpose of this study is to find out that if antibiotic streptomycin decreases neuronal network activity or affects the pharmacological responses. The experiments in this study were conducted via MEA (multi-electrode array) technology which records neuronal activity from devices that have multiple small electrodes, serve as neural interfaces connecting neurons to electronic circuitry. The result of this study shows that streptomycin lowered the spike production of neuronal network, and also, sensitization was seen when neuronal network pre-exposed to streptomycin.
Date: December 2014
Creator: Zeng, Wei Rong

Immunohistochemistry of the Gills of the Channel Catfish Ictalurus Punctatus: Cells and Neurochemicals That May Be Involved in the Control of Cardioventilatory Reflexes

Description: In teleost fishes the neurochemicals involved in sensing and responding to hypoxia are unresolved. Serotonergic branchial neuroepithelial cells (NECs) are putative O2 chemoreceptors believed to be homologous to the neural crest (NC) derived APUD (amine-precursor uptake and decarboxylation) pulmonary NECs and carotid body type-1 glomus cells. Branchial NECs contain serotonin (5-HT), thought to be central to the induction of the hypoxic cardioventilatory reflexes. However, application of 5-HT in vivo does not elicit cardioventilatory reflexes similar to those elicited by hypoxia. But previous in vitro neural recordings from glossopharyngeal (IX) afferents innervating O2 chemoreceptors in the trout gill show the same discharge response to hypoxic conditions as does that of acetylcholine (ACh) application. This evidence strongly supports the cholinergic hypothesis of chemoreceptor impulse origin rather than a serotonergic-induced impulse origin model. We therefore hypothesized that NECs contain ACh among other neurochemicals in cells belonging to the APUD series. Although serotonergic branchial NECs did not colocalize with ACh using immunohistochemical methods, several populations of ACh and/or tyrosine hydroxylase (TH) (catecholaminergic) positive, dopamine (DA) negative, cells were found throughout the second gill arch of the channel catfish Ictalurus punctatus. In addition, the NC derivation marker zn-12 labelled the HNK-1-like epitope (Human natural killer) expressed by lamellar pillar cells’ collagen column-associated pillar cell adhesion molecules (CC-PCAMs), evidence confirming their hypothesized NC origin.
Date: December 2013
Creator: Oden, David S.

Primary Cilia in the Oligodendrocyte Lineage

Description: oligodendrocytes migrate from the corpus callosum into the overlying cortex. The incidence of cilia did not change markedly across age groups, and did not vary consistently with the number of processes per cell, which was used as an indication of the maturation stage of OPCs and young OLs. The mean percent of Olig1 immunopositive (Olig1+) cells having cilia across ages was 33.1% + 16.5%, with all ages combined. In O4+ cells of these mice, 56.7 + 3.6% had primary cilia. If it is the case that adult OLs do not have cilia, the point in the lineage when primary cilia are lost is still unknown. Adult mice that had been injected with cyclopamine to block cilia-dependent Shh signaling were examined to determine whether the rate of generating new OPCs was influenced. In the CC of control mice, the numerical density of Olig1+/BrdU+ cells was 1.29 + 0.07/mm2 was reduced to 0.68 + 0.38/mm2 in the cyclopamine-injected group, and the numerical density of all BrdU+ cells (including both Olig1+ and Olig1- cells) of 4.55 + 1.50/mm2 in the control group was reduced to 3.14 + 1.27/mm2 in the cyclopamine-injected group. However, there were only 2 mice in each group and the differences were not statistically significant.
Date: May 2013
Creator: Hao, Yung-Chia

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 ...
Date: May 2006
Creator: Nebgen, Gilbert Bernard

Application of Cultured Neuronal Networks for Use as Biological Sensors in Water Toxicology and Lipid Signaling.

Description: This dissertation research explored the capabilities of neuronal networks grown on substrate integrated microelectrode arrays in vitro to be applied to toxicological research and lipid signaling. Chapter 1 details the effects of chlorine on neuronal network spontaneous electrical activity and pharmacological sensitivity. This study demonstrates that neuronal networks can maintain baseline spontaneous activity, and respond normally to pharmacological manipulations in the present of three times the chlorine present in drinking water. The findings suggest that neuronal networks may be used as biological sensors to monitor the quality of water and the presence of novel toxicants that cannot be detected by conventional sensors. Chapter 2 details the neuromodulatory effects of N-acylethanolamides (NAEs) on the spontaneous electrical activity of neuronal networks. NAEs are a group of lipids that can mimic the effects of marijuana and can be derived from a variety of plant sources including soy lecithin. The most prominent NAEs in soy lecithin, palmitoylethanolamide (PEA) and linoleoylethanolamide (LEA), were tested individually and were found to significantly inhibit neuronal spiking and bursting activity. These effects were potentiated by a mixture of NAEs as found in a HPLC enriched fraction from soy lecithin. Cannabinoid receptor-1 (CB1-R) antagonists and other cannabinoid pathway modulators indicated that the CB1-R was not directly involved in the effects of NAEs, but that enzymatic degradation and cellular uptake were more likely targets. The results demonstrate that neuronal networks may also be a viable platform for the elucidation of biochemical pathways and drug mechanisms of action.
Date: August 2004
Creator: Dian, Emese Emöke

Evaluation of virulence in wild type and pyrimidine auxotrophs of Pseudomonas aeruginosa using the eukaryotic model system Caenorhabditis elegans.

Description: 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.
Date: August 2004
Creator: Anvari, Sara

Chemotactic Response of Lumbricus terrestris Coelomocytes to Larval and Adult Stages of Rhabditis pellio

Description: 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.
Date: December 1999
Creator: Medrano, Jennifer Centurion

Evaluation of City of Denton Sub-Watershed by Benthic Macroinvertebrate Field Experimental Approach

Description: In this study, two different field experiments were designed to assess the relative influence of urbanization on benthic communities. During spring and summer, four urban and one reference sites from Denton County, Texas were selected for benthic macroinvertebrate evaluation. Statistically significant differences in colonized benthic macroinvertebrate taxa on artificial substrates were observed among the four urban sites and the reference site. Oligochaetes and chironomids were the dominant taxa at all sites. Identification of chironomid larvae at the subfamily and genus level to detect differences between sites had higher statistical power than the evaluation based on total chironomids. At the reference site, Caenis, Cladotanytarsus, Orthocladius, and Ceratopogonidae were the dominant taxa, while the urban sites were dominated by Dero, Physella, Ancylidae, Chironomus, Dicrotendipes, Glyptotendipes, Polypedilum, Pseudochironomus, Stenochironomus, and Tanytarsus. These differences may have been dependent upon differences in hydrologic regime and water quality between sites. Significant differences (ANOVA, p < 0.01) in water quality parameters (alkalinity, hardness, nitrates, phosphates, chlorides, sulfates, calcium, magnesium, potassium, and triazine) were found among water samples collected from the reference and urban sites. During the transfer period, most of the Ephemeroptera and Trichoptera taxa and a few other taxa disappeared from artificial substrates that were colonized at the reference site and then moved to the urban sites. Also, local abundant taxa from the urban site significantly (t test, p < 0.05) increased in number on the transferred artificial substrates. Seasonal differences in colonization patterns were also observed between the spring and summer experimental periods, which indicate that temporal variation is equally important, as is the anthropogenic effect in benthic community evaluation. Field survival and growth experiments using Erpetogomphus designatus larvae were designed to detect differences between evaluated sites. Larvae were collected from the reference site, measured in the laboratory, and exposed at the urban sites for ...
Date: August 2006
Creator: Mahato, Mahendra

A morphological study of the avian (Gallus domesticus) ductus arteriosi during hatching.

Description: The ductus arteriosi (DA) are two blood vessels connecting the pulmonary arteries to the descending aorta in the avian embryo. Following hatching, the DA closes, separation of the systemic and pulmonary circulation. I present the morphological changes that occur in the chicken DA during prepipping, internal pipping, external pipping, and hatching. The avian DA consists of two distinct tissue types, a proximal and a distal portion. Histological examination shows developmental differences between the proximal and distal portions of the DA with regard to lumen occlusion, endothelial cells, smooth muscle and elastin. Endothelial cell proliferation begins to occur as early as external pipping, with the lumen almost completely occluded by the 3rd day of post-hatching life. Expression of vascular endothelial growth factor (VEGF) increases in avian endothelial cells during hatching. I provide a morphological timeline of changes in the DA as the chicken develops from embryo to hatchling.
Date: May 2006
Creator: Belanger, Candace

Determination of Dissociation Constants for GABAA Receptor Antagonists using Spontaneously Active Neuronal Networks in vitro

Description: Changes in spontaneous spike activities recorded from murine frontal cortex networks grown on substrate-integrated microelectrodes were used to determine the dissociation constant (KB) of three GABAA antagonists. Neuronal networks were treated with fixed concentrations of GABAA antagonists and titrated with muscimol, a GABAA receptor agonist. Muscimol decreased spike activity in a concentration dependent manner with full efficacy (100% spike inhibition) and a 50% inhibitory concentration (IC50) of 0.14 ± 0.05 µM (mean ± SD, n=6). At 10, 20, 40 and 80 µM bicuculline, the muscimol IC50 values were shifted to 4.3 ± 1.8 µM (n=6), 6.8 ± 1.7 µM (n=6), 19.3 ± 3.54 µM (n=10) and 43.5 µM (n=2), respectively (mean ± SD). Muscimol titration in the presence of 10, 20, 40 µM of gabazine resulted in IC50s values of 20.1 (n=2), 37.17 (n=4), and 120.45 (n=2), respectively. In the presence of 20, 80, and 160 µM of TMPP (trimethylolpropane phosphate) the IC50s were 0.86 (n=2), 3.07 (n=3), 6.67 (n=2) µM, respectively. Increasing concentrations of GABAA antagonists shifted agonist log concentration-response curves to the right with identical efficacies, indicating direct competition for the GABAA receptor. A Schild plot analysis with linear regression resulted in slopes of 1.18 ± 0.18, 1.29 ± 0.23 and 1.05 ± 0.03 for bicuculline, gabazine and TMPP, respectively. The potency of antagonists was determined in terms of pA2 values. The pA2 values were 6.63 (gabazine), 6.21 (bicuculline), and 5.4 (TMPP). This suggests that gabazine has a higher binding affinity to the GABAA receptor than bicuculline and TMPP. Hence, using spike rate data obtained from population responses of spontaneously active neuronal networks, it is possible to determine key pharmacological properties of drug-receptor interactions.
Date: December 2005
Creator: Oli-Rijal, Sabnam

Pyrimidine Enzyme Specific Activity at Four Different Phases of Growth in Minimal and Rich Media, and Concomitant Virulence Factors Evaluation in Pseudomonas aeruginosa

Description: Pseudomonas aeruginosa is a Gram-negative rod, aerobic, non-fermenting, oxidase positive, pigment producing, and nutritionally versatile bacterium. Infections by P. aeruginosa are the most important cause of morbidity and mortality in immunocompromised patients, given virulence factor production that suppresses antibiotic therapy and promotes persistent infection. This research is the first comprehensive report of the pyrimidine biosynthetic pathway for all phases of growth in minimal and rich media coupled with the evaluation of virulence factor production of P. aeruginosa in comparison to four other bacterial species (Pseudomonas putida, Pseudomonas fluorescens, Burkholderia cepacia, and Escherichia coli wild-type strains). Cellular growth and passing genetic information to the next generation depend on the synthesis of purines and pyrimidines, the precursors of DNA and RNA. The pyrimidine biosynthetic pathway is essential and found in most organisms, with the exception of a few parasites that depend upon the pyrimidine salvage pathway for growth. Both the pyrimidine biosynthetic and salvage enzymes are targets for chemotherapeutic agents. In our laboratory, research on pyrimidine auxotrophic mutants showed the role of the pyrimidine biosynthetic pathway and its intermediates on P. aeruginosa metabolism and impaired virulence factors production. The present research shows that pyrimidine enzymes are active in all phases of growth, including the production of two forms of ATCase in the late log phase in P. aeruginosa. This finding may be explained by the displacement of the inactive PyrC' by the active PyrC or PyrC2 to form a new and larger pyrBC encoded ATCase. Pseudomonas aeruginosa wild-type appears to produce by far the most virulence factors, haemolysin, iron chelation, rhamnolipid, adherence, and three types of motility (swimming, swarming, and twitching) investigated in this study, when compared to the other four wild-type strains. Growth analysis was carried out as typically done in minimal medium but also in rich medium to simulate conditions ...
Date: December 2005
Creator: Azad, Kamran Nikkhah

Biogeography of Montane Mammals on the Colorado Plateau and Adjacent Regions

Description: This study identifies the biogeographic factors that structure small mammal communities on mountains of the Colorado Plateau and adjacent regions. Forty six isolated ranges were characterized across a 5-state study area encompassing the Colorado Plateau, including the central high plateaus of Utah and the Basin and Range Province (i.e. the Great Basin and mountains of Arizona and New Mexico). Presence/absence data of 25 montane mammal species were used to explore the interactions between historical and ecological processes affecting local and regional diversity patterns. Multivariate analyses, such as non-metric dimensional scaling, were used to explore factors which influence community composition. Results of these analyses revealed the Colorado River as a significant biogeographic barrier that affects montane mammal community structure. MtDNA cytochrome b sequence variation was analyzed among populations of the long-tailed vole, Microtus longicaudus, sampled from five interior ranges of the Colorado Plateau- Abajo, LaSal, Henry, and Chuska Mts., and Boulder Mountain of the Aquarius Plateau-and analyzed using traditional phylogenetic approaches (parsimony and likelihood) as well as nested clade analysis. Results support previous documentation of a major east-west phylogeographic break occurring between populations southeast of the Colorado River (eastern Arizona, Colorado, Wyoming and New Mexico) and all other western populations, which include a central clade, a northwest clade, and an Alaskan island clade. Evidence also supports differentiation of a 'southern Rockies' clade and a distinct 'southwest island' clade. Populations of M. longicaudus north and west of the Colorado River (Boulder and Henry Mts.) share two haplotypes, form a well-supported subclade with populations from the Kaibab plateau, and are closely related to the Northwest clade. Past approaches to studying montane mammal communities utilizing theory based on island biogeography have overemphasized area and isolation as the only forces structuring insular communities. As a result, there has been a lack of recognition of the ...
Date: May 2004
Creator: Carr, Carla B.

Utilization of Corridor Habitat by White-tailed Deer (Odocoileus virginianus) in Denton County, Texas

Description: 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.
Date: May 2004
Creator: Bruce, Troy Kenneth

Integrating Concepts in Modern Molecular Biology into a High School Biology Curriculum

Description: More so than any other science in the past several decades, Biology has seen an explosion of new information and monumental discoveries that have had a profound impact on much more than the science itself. Much of this has occurred at the molecular level. Many of these modern concepts, ideas, and technologies, as well as their historical context, can be easily understood and appreciated at the high school level. Moreover, it is argued here that the integration of this is critical for making biology relevant as a modern science. A contemporary high school biology curriculum should adequately reflect this newly acquired knowledge and how it has already has already begun to revolutionize medicine, agriculture, and the study of biology itself. This curriculum provides teachers with a detailed framework for integrating molecular biology into a high school biology curriculum. It is not intended to represent the curriculum for an entire academic year, but should be considered a significant component. In addition to examining key concepts and discoveries, it examines modern molecular techniques, their applications, and their relevance to science and beyond. It also provides several recommended labs and helpful protocols.
Date: August 2003
Creator: Parker, Timothy P.