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Roles Of Seed Dispersal And Environmental Filters In Establishment Of The Dominant Shrubs: Morella Cerifera And M. Pensylvanica, On An Atlantic Barrier Island, Benjamin Dows

Theses and Dissertations

Patterns of the expansion of woody cover into grasslands on barrier islands of the Virginia coast were investigated. Seed dispersal of the dominant shrub Morella spp., was sampled deploying seed traps (n = 82) throughout a landscape under shrub encroachment pressure on Hog Island, VA. Traps were placed underneath: fruiting Morella, non-fruiting Morella, co-occurring species (Iva frutescens and Baccharis halimifolia) and in grass land, (no shrub cover). Environmental filters that act upon dispersed seeds and subsequently determine establishment patterns were also investigated. Dispersal distribution throughout the encroachment zone was leptokurtic and dispersal among cover types suggest co-occurring shrub species facilitate …

Approaches To Reduce Selection Of Genomic Variants In Human Pluripotent Stem Cell Culture, Marion Riggs

Theses and Dissertations

Optimizing culture conditions that reduce genomic instability in human pluripotent stem cells (hPSCs) is an unmet challenge in the field. Results from our lab and numerous research groups demonstrate that hPSCs are prone to genomic aberrations and single-cell passaging increases the rate of genomic alterations. However, single-cell based passaging maintains advantages for scale-up and standardizing differentiation protocols. In this study, we investigated the problem of genomic instability in hPSC cultures with the goal towards identifying and characterizing candidate genes that could contribute to generation and survival of abnormal hPSCs. Based on microarray analysis, we identify ARHGDIA, located on 17q25, as …

A Passive Acoustic And Experimental Study Of Juvenile Blue Catfish, Ictalurus Furcatus, Sound Production And Agnostic Behavior In The Tidal Freshwater James River, Laura Morgan

Theses and Dissertations

Blue catfish, Ictalurus furcatus, are an invasive species in the James River, VA. They produce stridulation sounds and passive acoustic monitoring may prove useful in locating and monitoring their populations. Little is known about their behavior, therefore my goal was to examine agonistic behavior and the use of sound in defending a territory. This thesis consists of two manuscripts: 1) A passive acoustic study of the tidal freshwater James River, 2) An experimental study of agonistic behavior in juvenile Blue catfish, Ictalurus furcatus. The first study showed that three sounds (click, run croak) occurred more often in warmer months than …

Understanding The Role Of Stylochus Ellipticus As A Predator Of Crassostrea Virginica In Chesapeake Bay Tributaries, Marion Kensey Barker

Theses and Dissertations

Predation may be a key component of the unsuccessful restoration of the Eastern Oyster (Crassostrea virginica), a former keystone species in Chesapeake Bay. Here, I examine the polyclad flatworm Stylochus ellipticus and its potential role as an important predator of C. virginica. Using small-fragment size C. virginica specific DNA primers, oyster DNA was successfully detected in whole organisms homogenates of wild-caught S. ellipticus individuals. Of the 1,575 individuals tested, 68.1% tested positive, thus predation occurred. Predation did not appear to be affected by salinity or temperature; however, season did appear to have an effect on both predation and S. ellipticus …

Planar Cell Polarity And Neurodevelopment, Simon Sun

Theses and Dissertations

Planar cell polarity (PCP) is a developmental signaling mechanism that establishes a polarity within the plane of an epithelium. PCP has been shown to play a role in guiding numerous neurodevelopmental processes such as convergent extension, neuron migration, and axon pathfinding. Certain commissural neurons in the dorsal spinal cord make a series of guidance decisions en route to the brain: first, a ventral projection along the D-V axis, followed by a midline crossing, and after exiting the floorplate, a dorso-anterior turn along the A-P axis. Here, we provide in vivo evidence that the axons of the Commissural Primary Ascending (CoPAs) …

Conspecific Attraction In A Low-Density Population Of A Threatened Songbird, Daniel James Albrecht-Mallinger

Theses and Dissertations

Many organisms use both vegetation structure and social cues in selecting habitats. Many species of songbirds use the presence of breeding conspecifics as a social cue and sign of habitat quality, and can be induced to settle in unoccupied habitats by artificially broadcasting breeding song, a process referred to as “conspecific attraction”. In our study, we tested response to conspecific attraction a low-density population of the threatened Golden-winged Warbler (Vermivora chrysoptera) in Highland County, VA. Response to broadcast song was observed, with a modest increase in mean male abundance at survey points within 250 meters of treatments, and mean abundance …

Benthic Macroinvertebrate Subsampling Effort And Taxonomic Resolution For Bioassessments Of Streams In The James River Watershed Of Virginia, Laurel Williams

Theses and Dissertations

Benthic macroinvertebrate diversity influences stream food web dynamics, nutrient cycling and material exchange between the benthos and the water column. Stream bioassessment has moved to the forefront of water quality monitoring in terms of benthic macroinvertebrate diversity in the recent past. The objectives of this study were to determine optimum subsample size and level of taxonomic resolution necessary to accurately and precisely describe macroinvertebrate diversity in streams flowing in the Piedmont province of the James River watershed in Virginia. Forty-nine sampling sites were selected from streams within the Piedmont Physiographic Province of the James River watershed. Ten sites were randomly …

Indirect Effects Between Deer, Mice, And The Gypsy Moth In A Forest Community, John Wojcikiewicz

Theses and Dissertations

White-tailed deer are ecosystem engineers that dramatically alter forest understory vegetation. Consequently, deer can impact many species in a forest through both direct and indirect effects. One species that deer may indirectly affect is the gypsy moth, whose pupae are preyed upon by the white-footed mouse. Through alterations to understory habitat of mice, deer may reduce mouse predation on gypsy moth pupae. In this study, I tested for indirect effects of deer on the gypsy moth by comparing mouse abundance, vegetation properties, and predation on pupae inside, and outside, of long-term deer exclosures. Overall, I did not find evidence for …

Forest Net Primary Production Resistance Across A Gradient Of Moderate Disturbance, Ellen Goodrich-Stuart

Theses and Dissertations

The global carbon (C) balance is vulnerable to disturbances that alter terrestrial C uptake and loss. Moderate disturbances that kill or defoliate only a subset of canopy trees such as insect defoliation, drought, and age-related senescence are increasing in extent and frequency; yet, little is known about the effect of moderate disturbance on forest production and the mechanisms sustaining or supporting the recovery of the C cycle across a range of moderate disturbance severities. We used a broad plot-scale gradient of upper canopy tree mortality within a large manipulation of forest disturbance to: 1) quantify how aboveground wood net primary …

Causes And Consequences Of Algal Blooms In The Tidal Fresh James River, Joseph Wood

Theses and Dissertations

This dissertation includes 3 chapters which focus on algal bloom of the tidal fresh James River. The first chapter describes nutrient and light limitation assays performed on algal cultures and draw conclusions about long-term patterns in nutrient limitation by comparing results with a previous study . This chapter also describes the influence of riverine discharge upon nutrient limitation in a point-source dominated estuary. This chapter was published in Estuaries and Coasts (Wood and Bukaveckas 2014). The second chapter presents the first comprehensive assessment of the occurrence of the cyanotoxin Microcystin in water and biota of the James River. Data presented …

The Occurrence And Consequences Of Conspecific Brood Parasitism In The Prothonotary Warbler (Protonotaria Citrea), Anna Tucker

Theses and Dissertations

Brood parasites avoid costs associated with raising young by adding eggs to another individual’s clutch and providing no parental care. When conspecific brood parasitism occurs in species with high parental investment, we expect hosts to suffer costs for raising an enlarged brood. Here I describe conspecific brood parasitism (CBP) in the prothonotary warbler using maternal exclusion analyses of 333 family groups. I found that 23.4% of clutches contained at least one offspring that was not matched to the social mother and determined that parasitism seems to be an opportunistic tactic. Hosts had lower average annual reproductive success than non-hosts, but …

Hcpr Of Porphyromonas Gingivalis Utilizes Heme To Bind No, Benjamin Belvin

Theses and Dissertations

The obligate anaerobe Porphyromonas gingivalis is the etiological agent responsible for periodontal disease. It must withstand high levels of reactive nitrogen species in the oral cavity generated by the host and other oral flora. The mechanisms allowing for protection against such stress remain poorly understand. HcpR is an FNR-CRP family regulator that has been implicated in regulation of the nitrosative stress response. In this study we characterize the biochemical properties of HcpR. It is a homo-dimer that is composed of 3 domains – a heme-binding domain, dimerization helix, and a DNA-binding domain. Our studies show that HcpR binds the heme …

Forest Edges Enhance Mate-Finding In The European Gypsy Moth, Lymantria Dispar, Lily Thompson

Theses and Dissertations

Understanding movement capabilities of individuals within a landscape is essential to identifying the effects of habitat boundaries on species abundances, ranges, and spread rates. Movement barriers due to habitat fragmentation may reduce mate-finding ability in some species, particularly in heterogeneous landscapes containing low-density populations. This study focuses on the effects of habitat type and edge on mate-finding in an invasive defoliator, the European gypsy moth. Adult European gypsy moth males locate mates by following pheromones released by flightless females. Reduced mate-finding was expected in fields and near forest edges based on geographic variation in invasion rates and pheromone plume dynamics. …

Increased Circulatory Lipopolysaccharide From A High Fat Diet Aggravates Inflammation And Exacerbates Renal Failure, Samuel Righi

Theses and Dissertations

Kidney failure is frequently associated with the risk factors linked to metabolic syndrome. Lipopolysaccharide (LPS) is a potent inflammatory molecule, which has increased absorption from the gut into blood circulation following a high fat and high-energy diet. We hypothesized that LPS from a high fat diet can amplify inflammation, thereby exacerbating chronic kidney disease and associated disorders. We have found that adding a high fat diet to renal insufficient mice significantly progressed their kidney disease as well as associated disorders, compared to both a high fat diet and renal insufficiency alone. Additionally, we were able to demonstrate in vitro that …

Ampk Activators Regulate Contractile Function Of Mesenteric Arteries, Victoria Locke

Theses and Dissertations

Vasoconstrictor tone in the splanchnic circulation redistributes blood flow during hemorrhage and resuscitation. A metabolic sensor, 5’adenosine monophosphate-activated protein kinase (AMPK), has been proposed to relax arteries by inhibiting myosin light chain (MLC) kinase (MLCK) and rho kinase (ROCK) activities. Because AMPK activation might be beneficial in re-establishing splanchnic blood flow during resuscitation, we sought to explore the relative ability of AMPK activators (AICAR, A769662, berberine (BBR) and simvastatin (SIMV)) to relax mesenteric artery (MA) contraction. Our data revealed that these drugs caused vasorelaxation when tissues were stimulated either with KCl (producing primarily a Ca2+ dependent contraction) or phenylephrine (PE; …

Hydroxocobalamin Treatment For Carbon Monoxide Exposures: Characterizing Hemoglobin Changes And Testing For Neurological Sequelae, Leonardo Somera

Theses and Dissertations

Prior work in our lab has indicated that reduced Hydroxocobalamin (B12r) can be added to human blood and is able to convert carbon monoxide (CO) into carbon dioxide. This has great potential as a direct antidote to mitigate the toxic effects of CO poisoning which is a public health risk. In the first part of our work, we use highly specific wavelengths of light and Raman spectroscopy to study changes in Carboxyhemoglobin (COHb) between blood treated with oxygen and blood treated with oxygen and B12r in a flowing circuit of blood. Using Raman spectroscopy, we found that the addition of …

The Effect Of Retinoic Acid On Rai1 And Identification Of Retinoic Acid Receptor Binding Site In Human Rai1, Bingjie Xue

Theses and Dissertations

Previous studies showed haploinsufficiency of RAI1 is the main cause of Smith-Magenis syndrome (SMS). SMS is a developmental neurobehavioral syndrome characterized by intellectual disability, congenital anomalies, obesity, neurobehavioral abnormalities, and disrupted circadian sleep-wake pattern. SMS is caused by deletion or mutation of chromosomal region 17p11.2 that includes RAI1. Studies in the Elsea lab have shown that RAI1 is a dosage-sensitive gene. Haploinsufficiency of RAI1 leads to dysregulation of CLOCK, NR1D2, POMC, and BDNF, which are responsible for circadian rhythm, metabolism, and cognitive development. Based on the data from Elsea’s recent study on zebrafish, rai1 gene expression in zebrafish is regulated …

Structural Studies Of Interferon Regulatory Factor 4: A Molecular Perspective Of Its Regulatory Mechanism, Soumya Govinda Remesh

Theses and Dissertations

Interferon (IFN) regulatory factor family member 4 (IRF4) is a transcription factor that serves specific roles in transcriptional regulation of IFN responsive genes and is indispensable in B- & T-cell differentiation. IRF4 like the other members of the family has two major domains- the N-terminal DNA binding domain (DBD) essential for its recognition and binding to the Interferon Stimulated Response Element DNA sequence and a C-terminal Interferon activation domain (IAD) thought to maintain IRF4 in an auto-inhibited inactive state and is also critical in its activation. A putative unstructured linker connects the DBD and IAD. Activation in most members of …

Identification Of Micro-Rnas And Their Messenger Rna Targets In Prostate Cancer And Biological Fluids, Kanika Sharma

Theses and Dissertations

Prostate cancer is the second most common cancer in the United States that affects men today. To better treat this disease accurate biomarkers and successful therapeutic treatments are needed. A novel approach to understand the mechanisms behind prostate cancer tumor formation lies in identifying dysregulated micro-RNAs (miRNAs), which are a class of small (18-24 nucleotides) non-coding RNAs that regulate gene expression posttranscriptionally by either inhibiting protein synthesis or signaling messenger-RNA for degradation. Multiple miRNAs were discovered in our highly tumorigenic and metastatic prostate cancer progression model M12 cell line compared to its weakly tumorigenic P69 parental cell line. Various analyses …

Evolution And Divergence Of The Structural And Physical Properties Of Dna Binding By Methyl-Cytosine Binding Domain Family Members 2 And 3, Jason Cramer

Theses and Dissertations

The studies presented in this dissertation, Evolution And Divergence Of The Structural And Physical Properties Of DNA Binding By Methyl-Cytosine Binding Domain Family Members 2 And 3, pertain primarily to two key epigenetic regulators involved with the biological interpretation of methylated DNA marks. We provide insights into the emergence and evolution of the MBD2 and MBD3 and how those molecular entities influence heritable changes in gene activity. We further provide details regarding the mystery surrounding MBD3 function and the MBD2-mediated capacity of primitive animals to carry out methylation-specific epigenetic mechanisms. In chapter two, we describe the DNA binding properties of …

Developing C. Elegans As A Model To Study Type 2 Diabetes Mellitus, Jheesoo Ahn

Theses and Dissertations

Caenorhabditis elegans has been studied as a model organism in various areas of biomedical research because it shares many conserved functions at molecular and genetic levels with humans. Specifically, it is an ideal organism to study heterogeneous metabolic syndromes such as Type 2 Diabetes Mellitus (T2DM) as C. elegans can be used to delineate molecular pathways that are at the core of its problems. A growing number of populations worldwide are faced with chronic T2DM, which also manifests several complications, such as blindness, neuropathy and cardiovascular diseases. Currently, metformin is the first-line drug of choice administered to treat T2DM. While …

Voltage Sensing Mechanism In The Voltage-Gated And Proton (H+)-Selective Ion Channel Hv1, Aaron L. Randolph

Theses and Dissertations

Activation of the intrinsic aqueous water-wire proton conductance (GAQ) in Hv1 channels is controlled by changes in membrane potential and the transmembrane pH gradient (ΔpH). The mechanism by which changes in ΔpH affect the apparent voltage dependence of GAQ activation is not understood. In order to measure voltage sensor (VS) activation in Hv1, we mutated a conserved Arg residue in the fourth helical segment (S4) to His and measured H+ currents under whole-cell voltage clamp in transfected HEK-293 cells. Consistent with previous studies in VS domain containing proteins, we find that Hv1 R205H mediates a robust resting-state H+ ‘shuttle’ conductance …

A Systems Biology Approach To Detect Eqtls Associated With Mirna And Mrna Co-Expression Networks In The Nucleus Accumbens Of Chronic Alcoholic Patients, Mohammed Mamdani

Theses and Dissertations

Alcohol Dependence (AD) is a chronic substance use disorder with moderate heritability (60%). Linkage and genome-wide association studies (GWAS) have implicated a number of loci; however, the molecular mechanisms underlying AD are unclear. Advances in systems biology allow genome-wide expression data to be integrated with genetic data to detect expression quantitative trait loci (eQTL), polymorphisms that regulate gene expression levels, influence phenotypes and are significantly enriched among validated genetic signals for many commonly studied traits including AD.

We integrated genome-wide mRNA and miRNA expression data with genotypic data from the nucleus accumbens (NAc), a major addiction-related brain region, of 36 …

Investigating The Effects Of Applied Electric Fields On Microglial Cell Behaviour, Eman Bani

Theses and Dissertations

As surveyors of the central nervous system (CNS), microglial cells play an integral part in the inflammatory response following traumatic injuries. Thus, they have been implicated in the limited capability of neurons to regenerate in the CNS. Additionally, the roles of endogenous electric fields in the regenerative process of neurons in the mammalian peripheral nervous system (PNS) or amphibian CNS have long been studied. Further, previous studies in our lab have shown that physiological electric fields are capable of directing behaviours in astrocytes and schwann cells. Therefore in this study, a BV-2 microglia cell line was utilized to investigate whether …

Biochemical And Structural Studies Of Aav-2 Rep68-Aavs1 Complex Assembly, Clayton Bishop

Theses and Dissertations

Multiple DNA transactions are at the center of almost all processes regulating the AAV life cycle. A common feature shared by all transactions is the binding of the large AAV Rep proteins Rep78/Rep68 onto DNA sites harboring multiple GCTC repeats. AAV mediated site-specific integration is contingent upon the formation of a productive complex between Rep78/Rep68 and the AAVS1 site located at chromosome 19. In order to understand the mechanistic details of the initial assembly process we carried out equilibrium binding experiments of Rep68 and its individual domains with a 42-mer AAVS1 site. Results show that although Rep68 binds AAVS1 with …

Multi-Species Gene Networks And Drosophila Ethanol Sedation, Arnavaz Kollah

Theses and Dissertations

Alcohol use disorders (AUDs) are major health issues with few known genetic explanations. This project used the fruit fly (Drosophila melanogaster) model to identify genes and gene networks that influence alcohol intoxication, a phenotype related to alcohol abuse in humans. We used bioinformatic tools to build gene networks based on 24 published Drosophila ethanol-responsive genes with human orthologs. We then assessed the role of these networks in ethanol sedation by testing two of the networks seeded on IP3K2, a gene that regulates calcium signaling, and CG14630, a gene involved in carnitine biosynthesis. We knocked down several genes in each of …

Identification Of Peptidases In Highly-Pathogenic Versus Weakly-Pathogenic Naegleria Fowleri Amebae, Ishan Vyas

Theses and Dissertations

Naegleria fowleri, a free-living ameba, is the causative agent of Primary Amebic Meningoencephalitis. Highly-pathogenic mouse-passaged amebae (Mp) and weakly-pathogenic axenically-grown (Ax) N. fowleri were examined for peptidase activity. Zymography and azocasein peptidase activity assays demonstrated that Mp and Ax N. fowleri exhibited a similar peptidase pattern. Prominent for whole cell lysates, membranes and conditioned medium from Mp and Ax amebae were the presence of an activity band of approximately 58kDa and 100 kDa bands susceptible to the action of cysteine and metallopeptidase inhibitors, respectively. Further roles of the peptidases during the invasion process were examined by in vitro invasion assays …

Historical Tidal Forest Composition And Contemporary Woody Recruitment Following Dam Removal From A Mid-Atlantic Coastal Plain Tidal Freshwater Wetland, Richard E. Ward Jr.

Theses and Dissertations

Tidal freshwater forest restoration after dam removal has been unexplored to date. This study elucidated pre-dam forest composition, as well as post-dam edaphic and microtopographical attributes and woody species recruiting along a narrow ecotone of a 29.3-ha tidal freshwater wetland. The ≈65-year-old historical forest (15 species, 200 stems ha-1) and ≈7-year-old contemporary forest (40 species and 11,009 stems ha-) community dominants were dissimilar (Fraxinus spp. vs. Liquidambar styraciflua, respectively). Pre-dam environmental conditions were unknown. Post-dam edaphic water content, organic matter, redox potential and microtopography differed significantly across tidal sites but were less variable in non-tidal sites. Shifts in the contemporary …

Comparison Of Two Different Sprint Interval Training Work-To-Rest Ratios On Acute Metabolic And Inflammatory Responses, Christopher R. Harnish

Theses and Dissertations

High intensity exercise is believed to yield greater results on health and human performance than moderate intensity exercise. Extensive research indicates that not only do high-intensity interval training (HIT) and sprint interval training (SIT) produce significant improvements in cardiovascular fitness and disease, they may be more effective at improving long-term metabolic function, including insulin sensitivity (Si), by producing more mitochondria. Moreover, compliance rates for HIT and SIT participation are reported to be the same or better than traditional moderate intensity exercise. Because lack of time is often cited as major hindrance to exercise participation, SIT is also seen as a …

Investigating The Role Of The Synaptic Transcriptome In Ethanol-Responsive Behaviors, Megan A. O'Brien

Theses and Dissertations

Alcoholism is a complex neurological disorder characterized by loss of control in limiting intake, compulsion to seek and imbibe ethanol, and chronic craving and relapse. It is suggested that the characteristic behaviors associated with the escalation of drug use are caused by long-term molecular adaptations precipitated by the drug’s continual administration. These lasting activity-dependent changes that underlie addiction-associated behavior are thought, in part, to depend on new protein synthesis and remodeling at the synapses. It is well established that mRNA can be transported to neuronal distal processes, where it can undergo localized translation that is regulated in a spatially restricted …