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Cardiovascular Fetal Programming in Quail (Colinus virginianus), An Avian Comparative Model: The consequences of early embryonic insults and how they affect subsequent life reflects the emerging concept of "fetal programming". The aim of this project is to study the effects of embryonic insults as they subsequently manifest themselves in adults, with emphasis on the heart and vasculature. My experiments establish that fetal programming operates on the bobwhite quail inducing similar changes as those observed in mammalians and other birds. The quail's fast development provides reliable data in a short period of time than other avian models (e.g. domestic chicken). Data on quail showed a correlation between egg mass and hatchling mass; where small eggs produce small hatchlings but a high mortality made it impractical as a stressor for this study. Hypoxia was used as a stressor during embryonic incubation, where it induced a low hatching weight in quail that was not observable in adult birds. Morphological measurements demonstrated an increased ventricular collagen content and reduced ventricular lumen in birds in adults incubated in hypoxia consistent with hypertension. The hematological analyzes showed few differences indicating organ remodeling instead of hematopoietic compensation. The assessment of vascular reactivity pointed out an impaired endothelium dependent relaxation commonly associated to hypertension in birds and mammals. Fetal programming could be a widespread response to an adverse prenatal environment in endotherms and the resulting data from this work contributes to our understanding of fetal programming in vertebrates and its long term consequences.
Cellular Biomarkers for Measuring Toxicity of Xenobiotics: Effects of PCBs on Earthworm Lumbricus Terrestris Coelomocytes: The research presented herein provides information on coelomocyte (leukocyte) collection, function and immunotoxicity from polychlorinated biphenyls (PCB) in the earthworm Lumbricus terrestris. Research was undertaken as part of an overall goal to develop a well-documented and scientifically valid non-mammalian surrogate immunoassay with the earthworm Lumbricus terrestris to assess immunotoxic potential of xenobiotics. The principal objectives were to: (1) Develop an extrusion model for analyzing immunotoxicity of xenobiotics; (2) determine if coelomocytes can be collected repeatedly without obvious harm to the earthworm or change in immune response of the coelomocytes harvested and (3) validate the response sensitivity profiles of a panel of biomarkers {differential and total cell counts, erythrocyte rosette (ER) and secretory rosette (SR) formation with, and phagocytosis of antigenic rabbit red blood cells} in earthworms after acute exposure to a known mammalian and L. terrestris immunotoxin, the PCB Aroclor 1254.
Ceramide Biosynthesis and NEET Proteins Impact Development, Function, and Maintenance of the Caenorhabditis elegans Germline: I used the C. elegans genetic model to examine the role of ceramide biosynthesis (sphingolipid pathway) and iron regulation and found that each process impacts germline development and function. Using a sphingolipid specific antibody mAb15B4, I found that sphingolipids are associated with germ granules (P granules) within C. elegans and zebrafish; thus, suggesting conservation of macromolecules associated with germ granules. Phenotype analysis of ceramide biosynthesis mutants in C. elegans revealed that this pathway is essential for normal germline function in the aging adult hermaphrodite; specifically, precocious germline senescence was observed. Furthermore, I found that disruption of ceramide biosynthesis, via the hyl-2 deletion mutation, negatively impacts mAb15B4 localization at the P granules. Through genetic suppression analysis, I determined that insulin signaling and lipid biosynthesis can modulate the mAb15B4 localization to P granules. Additional, phenotype analysis showed that ceramide biosynthesis dysfunction decreased fecundity, and led to germline structure defects and uterine tumors. Through suppression analysis, I determined that modulation of the insulin signaling pathway suppressed the precocious germline senescence due to ceramide biosynthesis dysfunction. Since the presence of uterine tumors is associated with reproductive senescence I concluded that ceramide biosynthesis has a role in germline maintenance in the aging of the germline (germline senescence). The other important fate of a germ cell is programmed cell death. Apoptosis, which occurs through a highly conserved molecular pathway, is a normal component of growth and homeostatic processes. I used C. elegans to gain a greater understanding of the cisd gene function. The C. elegans genome has three previously uncharacterized cisd genes which code for CISD-1 (homology to vertebrate mitoNEET/CISD1 and NAF-1/CISD2) and CISD-3.1 and CISD-3.2 (homology to vertebrate Miner2/CISD3). I determined that independent disruption of the cisd genes resulted in a significant increase in the number of cell corpses within the adult hermaphrodite germline. Genetic …
Changes in Heterozygosity Through Time in American Standardbred and American Saddlebred Horses (1960-1990): Observed and expected heterozygosity (H) levels for ten electrophoretic blood marker loci and expected H for seven red blood cell (RBC) anitgen/antibody loci were examined for 20 years in American Standardbred and 30 years in American Saddlebred horses. Standardbreds were classed by gait, Trotter and Pacer, and evaluated separately in most analyses. 4,404 Trotters and 12,271 Pacers were found to have statistically highly significant losses of mean total observed H through time for the ten electrophoretic loci (P<0.005), although in Trotters the loss was more extreme (P<<0.001). Loss of H in 5984 Saddlebreds was not significant (P=0.259). Correlations of RBC expected H through time showed decreases in all three groups.
Changes which Occur in Components C'3 and C'4 in Guinea Pig Complement after Injection of an Antigen: This thesis describes an experiment involving guinea pigs' blood and the changes that occurred in the serum with the injection of an antigen. The serum was analyzed in complement titration tests.
Characteristics of Primary Cilia and Centrosomes in Neuronal and Glial Lineages of the Adult Brain: 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 …
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 in the Archaebacterium Methanococcus Jannaschii: Asparate transcarbamoylase catalyzes the first committed step in the de novo synthesis of pyrmidine nucleotides UMP, UDP, UTP, and CTP. The archetype enzyme found in Escherichia coli (310 kDa) exhibits sigmodial substrate binding kinetics with positive control by ATP and negative control with CTP and UTP. The ATCase characterized in this study is from the extreme thermophilic Archaebacterium, Methanococcus jannaschii. The enzyme was very stable at elevated temperatures and possessed activity from 20 degrees Celsius to 90 degrees Celsius. M. Jannaschii ATCase retained 75% of its activity after incubation at 100 degrees Celsius for a period of 90 minutes. No sigmodial allosteric response to substrate for the enzyme was observed. Velocity substrate plots gave Michaelis-Menten (hyperbolic) kinetics. The Km for aspartate was 7 mM at 30 degrees Celsius and the KM for carbamoylphosphate was .125 mM. The enzyme from M. jannaschii had a broad pH response with an optimum above pH 9. Kinetic measurements were significantly affected by changes in pH and temperature. The enzyme catalyzed reaction had an energy of activation of 10,300 calories per mole. ATCase from M. jannaschii was partially purified. The enzyme was shown to have a molecular weight of 110,000 Da., with a subunit molecular weight of 37,000 Da. The enzyme was thus a trimer composed of three identical subunits. The enzyme did not possess any regulatory response and no evidence for a regulatory polypeptide was found, DNA from M. jannaschii did hybridize to probes corresponding to genes for both the catalytic and regulatory subunits from E. coli. Analysis of DNA sequences for the M. jannaschii ATCase genes showed that the gene for the catalytic subunits shares significant homology with the pyrB genes from E. coli, and maximum homology amongst known ATCase genes to pyrB from Bacillus. An unlinked gene homologous to E. coli pyrl …
Characterization of H+ Excretion in a Model Renal Epithelium: The cellular regulation of acidification and intracellular ph (pHi) was studied in the integument of Rana pipiens, a model renal epithelium. Acidification was enhanced by : (1) chronic metabolic acidosis, (2) high salinity adaptation, and (3) ibuprofen treatment.
Characterization of the OCT Plasmid-Encoded Mercury Resistance Genetic Locus in Pseudomonas putida: A 17.1 Kb genetic element encoding for mercury resistance (OCT-Hg^r) was shown to translocate from its original location on the OCT plasmid to the resistance plasmid, RPl, in Pseudomonas putida. Analysis of RPl-Hg^r recombinant plasmids revealed that insertion of mercury resistance genes into RPl could occur at a variety of sites, with all recombinants having common EcoRI restriction fragments of 9.4, 3.8, 2.3, and 1.6 Kb, derived from the insertion. Hybridization analysis suggested the existence of extensive homology between this insertion and the prototypic mercury resistance transposon, Tn501, as well as the location of a similar merA sequence. Although the overall size was shown to be quite different from Tn501, striking physical similarities are shared between these two elements.
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 Molecular Changes of Austrofundulus limnaeus As It Develops Towards and Enters Diapause II: Austrofundulus limnaeus is a species of annual killifish which inhabits ephemeral ponds in South America. The species is able to survive seasonally desiccating ponds due to their ability to produce robust embryos. The embryos of this species are capable of entering a developmental arrest, termed diapause II, which precedes the onset of drought. While in this arrested state embryos exhibit the greatest tolerance to anoxia of any characterized vertebrate at 25ºC. Furthermore, when raised at 30ºC, embryos escape the entrance to diapause II and go on to develop directly. Currently, little is known about the molecular mechanisms which induce and maintain this developmentally arrested state. In this study I have developed methods to analyze changes in histone modifications in the context of diapause II. Histone modifications were chosen due to their extreme conservation and well characterized role as modulators of gene expression in other systems. Results utilizing adapted immunobased assays show significant changes in the global amount of H3S10P, H3K27me and H3K4me, as the embryos progress from early embryogenesis through the exit of diapause. Additionally, it is revealed that there exists a degree of phenotypic plasticity with regards to the entrance into diapause II which is modulated by the environment (severe hypoxia 0.1% O2). This work builds a foundation for future histone modification studies and contributes the development of several tools to the field. This study contributes to a greater molecular understanding of the cue(s) which influence the remarkable phenomenon of obligate developmental arrest in a vertebrate embryo.
Chemical and Physiological Properties of an Antibiotic Produced by a Variant of the Penicillium Notatum-Chrysogenum Group: The present study of the production and properties of an antibiotic was undertaken in the hope that some useful information could be contributed to a long-range investigation program being carried on at North Texas State Teachers College. This program is concerned with the production of a useful antibiotic from a variant of the Penicillium notatum-chysogenum group of molds.
Chemical Composition of the Peptidoglycan of Vitreoscilla Stercoraria: The peptidoglycan layer of Vitreoscilla stercoraria, ATCC 15218, was isolated from intact cells after treatment with sodium lauryl sulfate (SLS) and digestion with Pronase. Amino acid and amino sugar content was analyzed and 67% of the total present was made up of glutamic acid, alanine, diaminopimelic acid (DAP), and glucosamine in a molar ratio of 1:1.7:1:0.7. Electron microscopy of the final peptidoglycan product showed a thin, delicately folded sacculus which exhibited a morphology different from that of the intact vegetative cells. Within these sacculi occurred electron-dense structures which were assayed and found to be poly- 3-hydroxybutyrate (PHB) granules. The final yield of peptidoglycan was 2.9% of the dry weight of the intact vegetative cell.
The Chemical Compounds Produced by Actinomycetes and Their Relation to Tastes and Odors in a Water Supply: The purpose of this problem was to determine the cause of tastes and odors in the Breckenridge water supply and to isolate and culture the organisms responsible for the offensive chemicals.
Chemotactic Response of Lumbricus terrestris Coelomocytes to Larval and Adult Stages of Rhabditis pellio: 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 Acceleration and Osteogenesis: The effect of excess gravity on bone-forming elements of the growing perichondrial shaft of embryonic chicks was investigated through the use of the transmission electron microscope and various cytochemical techniques.
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.
Cigarette Residues Affect Steroidogenesis in Cultured Y-1 Mouse Adrenal Tumor Cells: This study (1) quantitatively compared steroid production in cultured Y-l mouse adrenal tumor cells exposed to Camel and Carlton-smoke derived residues, and (2) localized the effects in the cell. Basal steroid production was increased by Camel residues but not by Carlton, while ACTH stimulation was interfered with by both residues. Camel basal stimulation was comparable to that of cAMP, and was abolished by Cytochalasin D. The stimulation was also comparable to that of cholera toxin, which activates adenyl cyclase. Results indicate that residue components dissolve in the membrane stimulating adenyl cyclase at a point similar to or before that utilized by cholera toxin for its stimulating effect.
Circulatory and Respiratory Responses to Cycle Ergometry at Different Pedal Rates: The effects of moderate workload exercise at different pedal rates on circulatory and respiratory parameters were studied. Five subjects performed seven discontinuous constant-load cycle ergometer tests of 30 minutes duration at pedal rates of 40, 50, 60, 70, 80, 90 and 100 rpm. Oxygen uptake and carbon dioxide production were determined by standard open circuit spirometry, while heart rate was recorded by electrocardiograph. The CO₂ rebreathing procedure was administered during the exercise bout in order to determine cardiac output. Blood pressure was determined for each test, and total peripheral resistance was calculated. The findings indicate that progressive increases in pedal frequency during discontinuous constant-load cycle ergometry produce progressive increases in cardiovascular, respiratory and metabolic responses and a decrease in gross exercise mechanical efficiency. The results indicate that oxygen uptake, cardiac output, heart rate, ventilation and arterial-venous oxygen difference increases curvilinearly as pedal rate increases, possibly as a result of increases in recruitment of muscle fibers and/or muscle groups. These findings suggest that circulatory and respiratory responses are due to "central command" which sets the basic efferent response pattern. However, this effector pattern is modulated by afferent input originating from the legs during progressive increases in pedal rate.
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.
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.
CO2 Transport and Acid-Base Status during Fluctuations in Metabolic Status in Reptiles: Reptiles can often experience perturbations that greatly influence their metabolic status (e.g., temperature, exercise, digestion, and ontogeny). The most common cause of fluctuations in metabolic status in post-embryonic reptiles is arguably digestion and physical activity (which will be further referred to as exercise). The objective of this thesis is to determine the mechanisms involved in CO2 transport during digestion, determine the mechanisms that allow for the maintenance of acid-base homeostasis during digestion, and observing the effect of an understudied form of exercise in semi-aquatic reptiles on the regulation of metabolic acidosis and base deficit. This dissertation provided evidence for potentially novel and under investigated mechanisms for acid-base homeostasis (e.g., small intestine and tissue buffering capacity; Chapters 3 & 4), while also debunking a proposed hypothesis for the function of an anatomical feature that still remains a mystery to comparative physiologist (Chapter 2). This thesis is far from systematic and exhaustive in its approach, however, the work accomplished in this dissertation has become the foundation for multiple distinct paths for ecologically relevant investigations of the regulation of metabolic acidosis/alkalosis in reptiles.
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.
Colchicine Reversibly Inhibits Electrical Activity in Arthropod Mechanoreceptors: Dendrites of cockroach tibial spine mechanoreceptors contain hundreds of free microtubules, which may have some relation to the generation of electrical activity. Deflection of a spine produces a train of action potentials. Continuous perfusion over a period of 4 hours results in no response decrement. Perfusion with 10mM colchicine reversibly inhibits the response within 5-7 minutes. Irreversible inhibition is produced by perfusion with 1mM vinblastine sulfate in perfusion solution containing 1% dimethyl sulfoxide. Deuterium oxide does not inhibit at concentrations less than 50%, nor does it counteract inhibition by 10mM colchicine. Colchicine may be affecting (1) intracellular microtubules, (2) membraneous tubulin, (3) other membrane components, or (4) axoplasmic transport of essential materials to the sensory dendrites.
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.
Colonial Variations of a Mucoid Strain of Staphylococcus Aureus: It was the purpose of the present investigation to demonstrate the selection of a variant of one strain of Staphylococcus aureus by altering certain environmental factors, and to study that variant as to biochemical activities and capsule formation.
Combined Effects of Polycyclic Aromatic Hydrocarbons and Ultraviolet Light on Benthic and Pelagic Macroinvertebrates: Crude oil commonly enters freshwater aquatic ecosystems as thin sheens forming on the water surface. Oil contains mixtures of toxic compounds called polycyclic aromatic hydrocarbons (PAHs), some of which are known to be photodynamic, increasing toxicity when combined with ultraviolet radiation. Benthic macroinvertebrate communities are commonly utilized as bioindicators, and as such rely on abundant data in literature concerning benthic macroinvertebrates' relative tolerances to a wide range of pollutants. A series of 10 plastic traps, half of which were filtered from UV radiation, were deployed in an urban pond for 27 days to determine colonization preferences of benthic macroinvertebrates to UV exposure. Results of this in situ experiment indicated that the majority of aquatic insects collected from traps inhabited the UV exposed treatment group, particularly the nonbiting midge, Chironomidae. A series of bioassays were then completed to investigate the sensitivities of a Chironomidae species to thin sheens of crude oil in the presence and absence of UV radiation. All bioassays were conducted using 10 day old Chironomus dilutus larvae cultured in the lab. The series of C. dilutus bioassays were all conducted under the same water quality parameters, temperatures, and oil sheen dosing methods, under a 16:8 photoperiod and exposed to 16 h UV per day. Five replicates (n = 20) were loaded into 350 mL glass crystallizing dishes and exposed to four treatments for 96 hours: no UV/with sheen, with UV/no sheen, both UV and oil sheen, and a control. Three assays with 175 mL water volume were completed, one with no sediment, one with silica sand, and one with fine sand. Sediment type had a significant effect on mortality (p < 0.0001), but significant effects of UV or PAHs were not found. Two more C. dilutus assays were completed with identical parameters as the latter two with sediment, …
Comparative biochemistry and genetic analysis of nucleoside hydrolase in Escherichia coli, Pseudomonas aeruginosa, and Pseudomonas fluorescens.: The pyrimidine salvage enzyme, nucleoside hydrolase, is catalyzes the irreversible hydrolysis of nucleosides into the free nucleic acid base and D-ribose. Nucleoside hydrolases have varying degrees of specificity towards purine and pyrimidine nucleosides. In E. coli, three genes were found that encode homologues of several known nucleoside hydrolases in protozoa. All three genes (designated yaaF, yeiK, and ybeK) were amplified by PCR and cloned. Two of the gene products (yeiK and ybeK) encode pyrimidine-specific nucleoside hydrolases, while the third (yaaF) encodes a nonspecific nucleoside hydrolase. All three were expressed at low levels and had different modes of regulation. As a comparative analysis, the homologous genes of Pseudomonas aeruginosa and P. fluorescens (designated nuh) were cloned. Both were determined to encode nonspecific nucleoside hydrolases. The nucleoside hydrolases of the pseudomonads exhibited markedly different modes of regulation. Both have unique promoter structures and genetic organization. Furthermore, both pseudomonad nucleoside hydrolase were found to contain an N-terminal extension of 30-35 amino acids that is shown to act as a periplasmic-signaling sequence. These are the first two nucleoside hydrolases, to date,that have been conclusively demonstrated to be exported to the periplasmic space. The physiological relevance of this is explained.
Comparative Chemistry of Thermally Stressed North Lake and Its Water Source, Elm Fork Trinity River: To better understand abiotic dynamics in Southern reservoirs receiving heated effluents, water was analyzed before and after impoundment in 330 ha North Lake. Macronutrients, metals, and chlorinated hydrocarbons were measured. Concentrations of nutrients and metals in sediments were quantified in this 2 yr study. River water prior to impoundment contained 16 times more total phosphorus, and supported 23 times more Selenastrum capricornutum cells in an algal assay than reservoir water. The reservoir has essentially no drainage and since evaporation is high, the concentrations of many dissolved solids have increased since the reservoir was filled in 1958. North Lake is now phosphorus limited. Apparently altered chemical equilibria have caused precipitation or adsorption of phosphorus with calcium and iron.
Comparative Morphology of Sensilla Styloconica on the Proboscis of North American Nymphalidae and Other Selected Taxa: Systematic and Ecological Considerations.: Sensilla styloconica on the proboscis of 107 species of North American and tropical butterflies were comparatively studied using the scanning electron microscope. Focus was on 76 species of North American Nymphalidae representing 45 genera and 11 subfamilies. Nomenclature for generalized and specific types of nymphalid sensilla is proposed. Written descriptions and micrographs are presented for each species studied. Morphological features were generally consistent for all or most species within genera and sometimes within subfamilies, with specified exceptions. Statistical analysis revealed significant differences for six of eight variables tested between two distinct feeding guilds of North American Nymphalidae. Average number, density, extent of proboscis coverage with sensilla, their total length, and shoulder spine length were all significantly greater in the non-nectar feeding guild than in nectar feeders, and may indicate adaptation for greater efficiency in feeding on flat surfaces. The greater frequency of apical shoulder spines in non-nectar feeders may represent adaptation for protection of sensory pegs from mechanical abrasion during feeding, or for anchoring the flexible proboscis tip to the surface. Correlation analysis revealed 9 out of 28 positive correlations in nectar feeders and 5 out of 28 in non-nectar feeders. Results of preliminary cladistic analysis were not considered to be meaningfully robust due to few available characters. The stylar characters identified in this study should be more useful in future analyses when included with characters from other lines of evidence. The presence of sensilla styloconica in all subfamilies of Nymphalidae, except Danainae, largely supports Ehrlich's (1958) higher classification concept for the family. The presence of less conspicuous sensilla in the Danainae, and other characteristics are presented as further evidence that they should be reconsidered for full family status. Sensilla styloconica in nymphalid butterflies appear to function as extensions that provide greater sensory reach during feeding. The role of these …
A Comparative Study of Egg Media in the Primary Isolation of Mycobacterium Tuberculosis: The primary purpose of this investigation is not only to improve the present technique of culture of Mycobacterium tuberculosis, but also to make a comparative study of the media use, namely, Veterans Administration modification of Trudeau's medium, Lowenstein's egg medium, and that developed by the author, in order to find which, if any, of these will most easily and effectively meet the needs of the hospital laboratory.
A Comparative Study of Four Genealogies to Determine Predisposition to Cancer: This study was made in an effort first, to determine the mode of transmission of the inheritance of a predisposition to cancer as revealed through the analyses of four separate human genealogies, and second, to make a comparative study of the genealogies to determine whether the mode of transmission is the same.
A Comparative Study of Passive Transfer Mechanisms of Tuberculin and Chemical Contact Delayed Hypersensitivities in the Guiea Pig: This study is concerned with a critical comparison of the passive transfer mechanisms of tuberculin and chemical contact hypersensitivities in the guinea pig by use of a four phase experimental approach.
A Comparative Study of the Bottom Fauna of four Texas Lakes: This thesis attempted to study the bottom productivity both qualitatively and quantitatively with reference to the distribution of bonthos. The study of the bottom fauna in large reservior lakes is a relatively new field. This work will give more information on an unknown field than previously existed, although its scope is not intend to be exhaustive.
A Comparative Study of the D Cells of Certain Mammalian Islets of Langerhans: The purpose of this study is to describe and compare the D cells of the islets of Langerhans in six different species of mammals.
Comparative Toxicity of Refuse-Derived Fuel Fly Ash on Two Species of Earthworms, Lumbricus terrestris and E. foetida, Using an Artificial Soil Exposure Protocol: Research estimated toxicity of refuse-derived fuel fly ash (RDF-FA) on two earthworms species, Lumbricus terrestris and Eisenia foetida. Specific objectives were to: (1) Compare their 14-day LC50s under light and dark conditions; (2) separate toxicity due to osmotic, pH and physical factors from that of heavy metal contaminants; (3) compare relative differences of artificial soil and commercial soil as exposure media for evaluating toxicity to earthworms. The 14-d LC50s for L. terrestris in dark and light were 57.0 and 48.34 % RDF-FA, and 59.25 and 41.00 % RDF-FA for E. foetida using artificial soil. All of the toxicity resulted from heavy metals within the RDF-FA. Using L. terrestris, the LC50s for artificial soil and commercial soil were 52.30 and 64.34%.
Comparative Toxicity Responses in Earthworms Lumbricus Terrestris and Eisenia Foetida to Cadmium Nitrate and Chlordane Using Artificial Soil and Filter Paper Exposures: This research compares LC50 and LD50 of earthworms, Lumbricus terrestris and Eisenia foetida exposed to cadmium nitrate and chlordane using 48-h contact filter paper (FP) and 14-d artificial soil (AS) protocols. Both LC50 and LD50 showed that chlordane was more toxic than cadmium in both species regardless of the exposure. The reference toxicant 2-chloroacetamide using the standardized 48-h FP exposure was used to assess the general response of the earthworm prior to toxicity experiments. A glucose test was developed as an internal standard to assess homogeneity of mixtures among both replicates and dilutions. Accuracy of dilutions is assessed by the slope of a regression line relating nominal dilution to observed internal standard concentration. Precision of replicate preparation is assessed by among replicate variance.
Comparative Ultrastructural Study between Young and Adult forms of Trypanosoma Lewisi: The purpose of this study was to examine the ultrastructure of both young (dividing form) and adult (stationary form) of Trypanosoma lewisi. Changes observed between the two forms of the organism were related directly to data obtained from previous comparative biochemical studies conducted on these forms of Trypanosoma lewisi.
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.
A Comparison of Exotic and Domestic Species in American Zoological Gardens and their Practical Educational Value: This thesis presents a comparison of zoos and zoological gardens in the United States, Europe, and Asia. After an examination of these entities, the appeal and educational value of particular species is determined.
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.
A Comparison of Predicted and Actual Trophic Status of Lake Ray Roberts, Texas Based on Chlorophyll A: Two years before impoundment, the trophic status of Lake Ray Roberts was predicted by applying the total phosphorus input into the Organization for Economic Cooperation and Development (OECD) eutrophication model. Predicted mean summer epilimnetic (MSE) chlorophyll a of Elm Fork arm, Isle duBois arm and Main Body were in the eutrophic category of the OECD model. Observed MSE chlorophyll a two years after impoundment of Elm Fork arm, Isle duBois and Main Body had not reached their predicted means and were at the mesotrophic-eutrophic boundary of the OECD model. Six years after impoundment, observed MSE chlorophyll a for Main Body, was closer to its predicted mean and in the eutrophic category of the OECD model. Six years after impoundment, Elm Fork arm was the most productive area of Lake Ray Roberts. Observed means of chlorophyll a, total phosphates, suspended solids and turbidity were often highest in the Elm Fork arm. Wastewater effluent from Gainesville and Valley View, TX, had an impact on productivity in Elm Fork arm.
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.
A Comparison of Specificity of Passive Transfer of Chemical Contact Hypersensitivity Following Different Methods of Sensitization: This thesis compares the specificity of passive transfer of chemical contact hypersensitivity following various methods of sensitization.
A Comparison of the Carbon Dioxide and Oxygen Rate of Change Methods for Measuring Primary Productivity: The purpose of this study was to demonstrate the variability of oxygen and carbon dioxide rate of change methods for measuring primary productivity and respiration in an aquatic environment.
A Comparison of Thermogenesis by Selected Substrates on Hypothermic Rat Liver: The thermogenic effects in hypothermia of four substrates--alanine, glycine, ethano, and pyruvate - were studied in seventeen experiments. Albino rats were decapitated, and their livers were removed. The livers were homogenized with phosphate buffer at -5° C. After equilibration in a refrigerated Warburg apparatus at 20° C, the substrates were added and tissue respiration was recorded over three hours. Heat production was calculated from O2 uptake and CO2 production. Results showed that alanine, glycine, and pyrvate yielded 93.19, 89.86, and 89.89 x 10^6 kg-cal compared to a control value of 86.11 x 10^-6 kg-cal. Ethanol provided 110.31 x 10^-6 kg-cal, a value significantly greater than for the other substrates. The substrates studied, especially ethanol, did, therefore increase heat production in an artificially hypothermic environment in homogenized rat livers.
Composition and Distribution of the Vegetation in Farm Pastures in Denton County, Texas: This investigation has for its aim the determination and the evaluation of the vegetational composition in certain farm pastures in Denton County, Texas.
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.
The Consequences of Early Life Stage Thyroid Suppression on Immune Function in the Fathead Minnow (Pimephales promelas): Current evidence suggests that thyroid hormones (THs) may impact development of the immune system. However, studies that explore the role of THs in immune development are limited, and the mechanisms leading to alterations in immune function are poorly understood. It is important to elucidate the role of THs in immune development given that many environmental contaminants have been shown to disrupt TH homeostasis and may also have negative impacts on the immune system. As such, the main goal of this study was to determine the long-term consequences of early life stage (ELS) hypothyroidism on immune function. To achieve this goal, it was first necessary to further characterize basic immune function in the selected model species, the fathead minnow (FHM, Pimephales promelas). Preliminary studies were conducted to describe the transcriptomic response to Yersinia ruckeri and adapt assays for the assessment of respiratory burst and phagocytic cell activity. To determine the long-term effects of ELS hypothyroidism, FHMs were exposed to the model thyroid suppressant propylthiouracil (PTU) from <1 to 30 days post hatch and reared under normal conditions. Upon reaching adulthood, ex vivo immune cell function and the in vivo immune response to Y. ruckeri were assessed. Fish exposed to PTU experienced significant alterations in gene networks associated with immune cell function as well as significant decreases in phagocytic cell activity. However, immune-related alterations at the molecular and cellular levels did not manifest themselves at higher levels of organization as ELS hypothyroidism did not affect any other immune-related endpoints, including pathogen resistance. To our knowledge, this is the first study to provide evidence that ELS hypothyroidism causes long-term effects on the immune system in fish.

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