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Visual Circuitry In Adaptive Behavior: Extraction Of Complex Visual Image Information By Arrays Of Non-Image Forming Eyes, Thomas Kipp Halladay Groves

Dissertations

A complete understanding of how neural circuitry underlies the detection and integration of visual cues continues to elude researchers in the 21st century. Since the dawn of life on earth, light has exerted enormous selective pressure on all living organisms. Visual systems represent one of the most common sensory systems in existence, with significant homologies among elements of these systems, such as photopigments and developmental genes. Offering nearly unmatched single cell access at all levels of its nervous system, the European medicinal leech (Hirudo spp.) is an excellent system in which to identify behaviorally relevant neuronal circuits. Hirudo evolved discrete …

From Young To Old: The Effects Of Sedentary-Aging And Exercise Interventions On Structural Plasticity Of Lumbar Motor Neurons, Nmj, And Glial Cell Line-Derived Neurotrophic Factor Expression, Alberto Felipe Cintron-Colon

Dissertations

Neuromuscular weakness has been associated with age progression. A possible reason is a disconnection between muscle and neuron, causing components from both the central and peripheral nervous systems to deteriorate. Exercise, however, has beneficial effects for both muscle and nervous tissue, inducing neuroprotection. Neurotrophic factors are powerful regulators of neuronal maintenance and synaptic strength. Glial cell line-derived neurotrophic factor (GDNF) is a neurotrophic factor that has been shown to be a potent survival factor for somatic motor neurons that innervate skeletal muscle.

This study aims to evaluate changes in structural plasticity of motor neurons in the lumbar spinal cord and …

The Impacts Of Aging, Sedentarism, And Exercise On Neurotrophic Factor Expression And Innervation In The Heart And The Effects Of Treatment With Α-Cgrp On Heart Function, Gabriel Almeida Alves

Dissertations

Neurotrophic factors (NFs) are important molecules responsible for development, differentiation, regeneration, and maintenance of new and mature neurons. Neurotrophic factors act as neurocytokines and may assist with the regulation of axonal and dendritic arrangements and synaptic plasticity between neurons themselves or with other non-neural target tissues. In this study, we analyze the levels of two NFs: glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF). Cardiomyocytes produce these neurotrophic factors which assist with the innervation pattern of the heart. The heart is innervated by the two branches of the autonomic nervous system; namely the sympathetic nervous system and …

Myosin Assembly Into Elongating Thick Filaments Is Dependent On The Function Of Conserved Apmk-Related Kinase Unc-82 In C. Elegans, Natasha R. Schiller

Dissertations

The mechanism of assembly of sarcomeric myosins into elongating thick filaments is not well understood. The results of this study suggest a novel mechanistic model in which the AMPK-related kinase UNC-82, which is orthologous to mammalian ARK5/NUAK1 and SNARK/NUAK2, acts as an assemblase on sarcomeric myosin molecules, mediating their addition into the elongating thick filaments of C. elegans body-wall muscle. The classical genetic crosses performed in this study produced over 53 different worm strains that contain thick-filament-affecting mutations and/or transgenes in single-, double- or triple-mutant combinations. The phenotypes of these strain were documented and analyzed using viability assays, brood analysis, …

Evolution Of Caffeine Biosynthetic Enzymes And Pathways In Flowering Plants, Ruiqi Huang

Dissertations

Convergent evolution generally refers to the independent evolution of similar biological function more than once in unrelated species. Caffeine is thought to have evolved by convergence, and is naturally produced through secondary metabolism in plants to defend against pathogen attack and insect feeding or to attract pollinators. The same caffeine biosynthetic pathway has been elucidated in Camellia (tea) and Coffea (coffee), in which xanthosine is sequentially methylated to caffeine via 7-methylxanthine and theobromine. However, although the same catalysis pathway is used, different (paralogous) enzymes in the SAMT/BAMT/theobromine synthase (SABATH) multigene family are used in the two species. In my dissertation, …

Oncolytic Tanapoxvirus For Melanoma Therapy, Tiantian Zhang

Dissertations

Oncolytic viruses (OVs), which preferentially infect cancer cells and induce host anti- tumor immune responses, have emerged as an effective melanoma therapy. Tanapoxvirus (TPV), which possesses a large genome and causes mild self-limiting disease in humans, is potentially an ideal OV candidate. The purposes of our studies are to engineer TPV into effective OVs via arming immumo-stimulatory proteins and/or manipulating the virokines, and to explore the immuno-modulatory activities of TPV.

Interleukin-2 (IL-2) plays a critical role in activating T cells, natural killer cells and macrophages in both the innate and adaptive immune systems. In our study, a recombinant TPV expressing …

Investigations Of Filarial Nematode Motility, Response To Drug Treatment, And Pathology, Charles Nutting

Dissertations

More than a billion people live at risk of chronic diseases caused by parasitic filarial nematodes. These diseases: lymphatic filariasis, onchocerciasis, and loaisis cause significant morbidity, degrading the health, quality of life, and economic productivity of those who suffer from them. Though treatable, there is no cure to rid those infected of adult parasites. The parasites can modulate the immune system and live for 10-15 years. Testing of compounds against filarial nematodes is complicated due to a lack of an objective platform on which to analyze in vitro treatments. There is no published, immunocompetent laboratory model for lymphatic filariasis. This …

Regulation Of Glial Cell Line-Derived Neurotrophic Factor (Gdnf) Production In Voluntary And Involuntary Muscle, John-Mary Vianney

Dissertations

Glia cell line-derived neurotrophic factor (GDNF) is a survival factor for subpopulations of neurons, including somatic and autonomic motor neurons. These neurons depend, in part, on GDNF that is synthesized and secreted by their target tissues. It has been shown that a number of tissues in the periphery express GDNF and these target tissues differ in their composition, function, and in the case of different muscle cell types, their contractile characteristics. Whether the processes regulating GDNF production in these different tissues is similar or different is poorly understood. The broad goal of this study is to examine factors that normally …

Population Demographics And Genetics Of Spix's Disk-Winged Bat: Insights Regarding Survival, Mate Choice, Gene Flow And Effective Population Size, Michael Buchalski

Dissertations

Simultaneous study of the demographics and genetics of populations are relatively rare within the literature, despite insights their combined use offers regarding the life history, ecology, and evolution of species. Here I take a comprehensive approach, using capture-recapture data, polymorphic microsatellite markers, and various modeling techniques to examine the demographics and genetics of multiple local populations of Spix’s disk-winged bat (Thyroptera tricolor) in southwestern Costa Rica. T. tricolor is a highly gregarious, neotropical bat species known to form kin-based social groups with high retention of offspring of both sexes. The implications of this highly unusual social structure for …

Discovery And Characterization Of A Tanapoxvirus Encoded Human Neuregulin Mimetic, David Jeng

Dissertations

Studies on large dsDNA viruses such as poxviruses have been helpful in identifying a number of viral and cellular growth factors that contribute to our broad understanding of virus-host interaction. Orthopoxviruses and leporipoxviruses are among the most studied viruses in this aspect. However, tanapoxvirus (TPV) a member of the genus Yatapoxvirus, still remains largely unexplored, as the only known hosts for this virus are humans and monkeys. Here, we describe the initial characterization of an EGF-like growth factor mimicking human neuregulin from TPV, expressed by the TPV-15L gene. Assays using a baculovirus expressed and tagged TPV-15L protein demonstrated the ability …

Neuroprotective Effects Of A Nicotinic Acetylcholine Receptor Agonist And Modulator In The Rodent Retina, Kazuhiro Iwamoto

Dissertations

The purpose of this dissertation is to assess the potential neuroprotective effect of an alpha7 nAChR agonist, PNU-282987, and modulator, PNU-120596, in an in vitro model of excitotoxicity and an in vivo model of acute glaucoma. In in vitro studies, retinas obtained from Long Evans rats were cultured using various concentrations of the PNU compounds to analyze neuroprotection against glutamate-induced excitotoxicity. After 3 days in culture, RGCs were identified using an antibody against Thy 1.1, visualized using a fluorescent dye and quantified. In culture, glutamate significantly decreased the number of RGCs. However, if either PNU compound was introduced before the …

Molecular, Cellular And Systemic Effects Of Atrazine On The Xenopus Laevis Tadpole, Renee Maryanne Zaya

Dissertations

Atrazine is one of the two most commonly used herbicides in the US and it is the most studies are among the first to link physiological effects of atrazine to changes in tissue and gene expression in Xenopus laevis tadpoles. These studies provide the groundwork for future studies into the mechanisms behind responses to chemical stressors generating data resulting in a better understanding of how these chemicals affect us all. commonly found herbicide in ground water. As a result, a great deal of attention has been placed on its use and safety. It has also been implicated to play a …

High-Throughput Screening Studies Of Inhibition Of Human Carbonic Anhydrase Ii And Bacterial Flagella Antimicrobial Activity, Albert A. Barrese Iii

Dissertations

Complete abstract attached as supplemental file.

Construction Of A New Model System In Saccharomyces Cerevisa To Study Yopo And Identify Its Targets, Laura Nejedlik

Dissertations

Yersinia species use a type three secretion system (TTSS) to inject Yersinia outer proteins (Yops) into the target cell. The effector YopO is a serine/threonine kinase which upon association with actin becomes active. However, activation of the kinase is not required for actin disruption. YopO also contains a Rho/Rac binding domain, but it is unclear how it affects these proteins.

We have established an expression system in Saccharomyces cerevisiae to study YopO. The expression vector contains a GAL1 promoter for inducible expression of YopO as well as a V5 epitope used for visualization of each Yop. We have shown that …

Characterization Of The Ap-1 And Nf-Kappa B Transcription Factors In The U-87 Mg Astrocytoma Cell Line, Denise L. Smith

Dissertations

Growth factors, signaling molecules and transcription factors are frequently mutated in cancer and platelet-derived growth factor (PDGF) has been shown to be over-expressed in most gliomas. In the astrocytoma cell line U-87 MG, the over-expression of PDGF has been shown to lead to over-expression of transcription factors activator-protein-1 (AP-1) and nuclear factor-kappa B (NF-κB), and it is believed that the individual subunits of AP-1 (c-Jun, JunB, JunD, c-Fos, FosB, Fra-1, and Fra-2) and NF-κB (c-Rel, p50, p52, p65, and RelB) play different roles in proliferation, survival and differentiation in the U-87 MG cell line.

To assess these different roles, transient …

Mapk Survival Signaling In Melanoma, Matthew W. Vanbrocklin

Dissertations

Extracellular signals activate mitogen-activated protein kinase (MAPK) cascades potentiating biological activities such as cell proliferation, differentiation, and survival. Constitutive activation of MAPK signaling pathways is implicated in the development and progression of many human cancers, including melanoma. Mutually exclusive activating mutations in NRAS or BRAF are found in ∼85% of all melanomas resulting in constitutive activation of the MAPK pathway (RAS-BRAF-MEK-ERK-RSK). We have previously demonstrated that inhibition of this pathway with small molecule MEK inhibitors selectively induces apoptosis in human melanoma cells both in vitro and in vivo, but not in normal melanocytes. These results support the notion that the …

The Effects Of Aging And Activity On Glial Cell Line-Derived Neurotrophic Factor Expression In Skeletal Muscle, Nathan Peplinski

Dissertations

Glial cell line-derived neurotrophic factor (GDNF) is an extracellular signaling protein that is produced by skeletal muscle and is important for the motor neurons that control muscle movement. GDNF has been shown to keep neurons alive under conditions that they would otherwise not persist. In skeletal muscle, GDNF has been shown to be one of the most potent neurotrophic factors that influence motor neuron survival. While the role of GDNF has been well studied during early development, not much is known about what happens to GDNF expression in the adult and with advanced aging. Previous results from our lab have …

Structure-Function Studies Of Self- Assembling Flagellin Proteins Fromsalmonella Typhimurium And Aquifex Pyrophilus, Venkata Raghu Ram Malapaka

Dissertations

The eubacterial flagellum is a complex structure with an elongated extracellular filament that is primarily composed of a single protein termed flagellin. The highly conserved N-and C- termini of flagellin are important in its export and assembly, whereas the middle "hypervariable" region is highly variable in size across different species and is largely dispensable. InSalmonella typhimurium phase 1 flagellin (FliC), this hypervariable region encodes two solvent-exposed D2 and D3 domains. The functional role of this knob-like structural feature in motility remains unclear. The structural and physiological role of the hypervariable region in flagellar assembly, stability and cellular motility was investigated. …

Characterization Of Membrane Potentials In Vascular Smooth Muscle Of Hagfish, Lamprey And Trout, Prentiss Jones Jr.

Dissertations

The objective of this study was to characterize membrane potentials in systemic arteries of Pacific hagfish, Eptatretus stouti, Rainbow trout, Onchorhyncus mykiss, and Sea lamprey Petromyzon marinus. Previous studies have characterized membrane potentials in piscine tissue. However, these studies utilized non-vascular tissues such as cardiac muscle and skeletal muscle (69, 161). Characterization of membrane potentials in fish vasculature is without precedent in the literature. The hypothesis of this study was that membrane potentials in fish vascular smooth muscle differ little between species when comparable vasculature is evaluated.

Molecular Analysis Of The Effects Of Atrazine On Xenopus Laevis Frogs, Ronald Edward Celestine

Dissertations

Atrazine, the most commonly used herbicide in the U.S., has been implicated as one of the factors that may be causing the worldwide decline of amphibians. First, I examined effects of atrazine on laboratory raised Xenopus laevis tadpoles, treated from 5 days post fertilization to NF stage 65/66 with a concentrationof 25 ppb of atrazine. This study showed no significant difference in body weight, length and metamorphic rate.

Second, I used DNA microarrays to examine mRNA expression levels of genes in atrazine treated and control Xenopus laevis tadpoles. I found significant changes in gene expression levels of 18 genes. These …

Exposure Of Nervous System Cells To Polychlorinated Biphenyls (Pcbs), Results In Alterations Of Neurotrophic Factor Expression, Gordon H. Gurley

Dissertations

Toxic insult by PCBs results in learning and memory deficits in humans. Alterations in expression of neurotrophic factors (NF) and/or their receptors have been linked to changes in learning and memory. How PCBs affect cognition is not known. We suggest that PCBs affect cognition by altering NF expression.

We exposed cultured C6 rat glioblastoma cells (an astrocyte cell-line) to medium containing PCB (Aroclor 1254 (10ppm)). Control cells were treated with dimethyl sulfoxide (DMSO) or regular medium. Enzyme-linked immunosorbent assays (ELISA) were used to determine glial cell-line derived neurotrophic factor(GDNF) and nerve growth factor (NGF) concentrations in all samples. We also …

Comparative Genomic Analyses And Expression Of A Set Of Overlapping Open Reading Frames Of Frog Virus 3, Type Species Of Ranavirus (Family Iridoviridae), Wendy Guat Hoon Tan

Dissertations

Frog virus 3 (FV3) is the type species member of the genus Ranavirus (family Iridoviridae ). To better understand the molecular mechanisms involved in the replication of FV3, including transcription of its highly methylated DNA genome, the complete nucleotide sequence of the FV3 genome has been determined. The FV3 genome is 105, 903 base pairs long excluding the terminal redundancy. The G+C content of FV3 genome is 55% and it encodes 98 non-overlappingpotential open reading frames (ORFs) containing 50 to 1293 amino acids. Eighty-four ORFs have significant homology to known proteins of other iridoviruses, whereas 14 are unique FV3 ORFs. …

Olfactory Bulb Neurons Of Adult Zebrafish Morphology, Distribution, Cellular Interactions And Structural Stability Following Deafferentation, Cynthia L. Fuller

Dissertations

The zebrafish is becoming an increasingly popular model for studies involving olfactory function, yet there is still much to be learned about the anatomy and circuitry of different cell types in the olfactory bulb. This study focuses on identifying the morphology and distribution of output neurons and interneurons in the olfactory bulb of adult zebrafish, Danio rerio . Furthermore, this investigation examines the cellular interactions of the primary output neuron, the mitral cell, and addresses the issue of neuronal plasticity by considering the structural stability of this cell type following loss of afferent innervation.

Using retrograde tract tracing with various …

Bufadienolides In The Chemical Defenses Of The Toads, Bufo Americanus And Bufo Fowleri, Catherine E. Merovich

Dissertations

Resource equalization represents the historical and theoretical basis for American school finance and its ideals continue to dominate discussion and practice. During the last decade and a half, the national aspiration for equitable distribution of school finance resources has started a transformation into a derivative notion of adequacy. This equity policy derivative embraces the idea of sufficiency, which in turn implies a standard. The concept of educational adequacy has grown to mean the achievement of high absolute standards. Equity (equalization) theorists measure resource level on a relative basis; each student relative to all others. Adequacy adherents measure student achievement against …

The Glucose And Insulin Responsiveness Of The Rat Glucose-6-Phosphate Dehydrogenase Promoter, Daryl Arkwright-Keeler

Dissertations

The regulation of expression of some enzymes involved in glucose homeostasis and fat metabolism requires both insulin and glucose to elicit an effect, while the regulation of others requires only one. Glucose-6-phosphate dehydrogenase (G6PDH), an important enzyme involved in glucose homeostasis, has been shown to be transcriptionally regulated by carbohydrate feeding in rat liver, a diet regime that increases the levels of both glucose and insulin. Our lab had previously shown that insulin induces expression of G6PDH, thus we wanted to determine if glucose also induced G6PDH expression and if so, are the glucose and insulin responses mediated through identical …

Changes In Glial Cell Line Derived Neurotrophic Factor And Nerve Growth Factor Expression With Development, Age, Exercise And Hypertension, Bertha C. Rebimbas-Cohen

Dissertations

Neurotrophic factors are important for neuronal growth, survival, and maintenance of cell phenotype. Glial cell line-derived neurotrophic factor (GDNF) affects sensory, autonomic and somatic motor neurons and nerve growth factor (NGF) affects sensory and sympathetic neurons. NGF and GDNF are produced by cardiac muscle and have potent effects on sensory and autonomic neuronal innervation of blood vessels. Neural innervation plays a key role in blood vessel function. Since neural innervation of blood vessels may be regulated by growth factors, we wanted to determine what changes would occur to NGF and GDNF levels with maturity, exercise and hypertension.

Changes in GDNF …

K+- Induced Dilation Of Cremasteric Arterioles Involves Na+/K+Atpase And Inward Rectifier K+ Channels, Wendy Burns

Dissertations

The purpose of this study was to evaluate the possible mechanisms involved in cremasteric arteriolar dilation as a result of increased concentrations of ([K⁺]_o) extracellular potassium. Many other studies suggested that Na⁺/K⁺ ATPase or K_IR channels are responsible for potassium-induced vasodilation in other systems (2, 12, 22, 40, 46, 52, 16, 17, 25, 33, 34, 85, 99, 151). However, these studies show only partial inhibition of dilation as a result of independent blockageof each mechanism. The hypothesis of this study was that both mechanisms contribute to potassium-induced dilation seen in cremasteric arterioles. …

Cellular Proliferation, Death And Histology In Gambusia Affinis Livers After Exposure To 2-Aminofluorene And Benzidine, Susan J. Lentz

Dissertations

A fundamental need exists for aquatic animals in the biomonitoring of aquatic ecosystems for environmental carcinogens. G. affinis, a small fish species, is selected for this study due to its hardiness, non-migratory nature, geographic diversity and ability to develop tumors. Molecular tools for cellular proliferation and death are examined for use with G. affinis. Additionally, these tools are used to investigate the early carcinogenic effects of 2-aminofluorene and benzidine, both individually and combined, on G. affinis livers.

Antibodies to proliferating cell nuclear antigen (PCNA), p53, cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase (PARP) were examined for use in G. …

Identification And Classification Of Coryneform Bacteria Isolated From Bovine Mammary Glands, Jeffrey L. Watts

Dissertations

Coryneform bacteria are frequently isolated from bovine mastitis and these infections are associated with economic losses. Corynehacterium bovis, a lipid requiring species, has been the most frequently isolated coryneform from the milk of infected bovine mammary glands. However, the taxonomic status of this organism is uncertain. In the current study, a polyphasic approach was used to identify coryneform bacteria isolated from bovine mastitis and determ ine the phylogenetic relationships among the identified species. A total of 212 coryneform bacteria isolated from bovine mastitis was obtained from mastitis reference laboratories in the United States and Canada. Presumptive identification based upon …

Molecular And Biological Characterization Of A Tanapox Virus Tnf-Binding Protein And The Complete Nucleotide Sequence Of The Psti-J Genomic Fragment, Mini Paulose-Murphy

Dissertations

Most of the poxvirus encoded virulence factors have been identified as proteins that are secreted from infected host cells. Some of these secretory proteins impede host immune defenses. We have previously demonstrated that tanapox virus (TPV) infected cells secrete a 38 kDa glycopeptide that binds to human (h) interferon-γ, h-interleukin (IL)-2, and hIL-5. We now show an additional activity in the supernatant from TPV infected cells that down-regulates the expression of tumor necrosis factor-a (TNF-α) induced cell adhesion molecule gene expression. This activity was not detected in mock infected cells. Enzyme linked immunosorbent assays (ELISA) on primary human endothelial cells, …