Life Sciences Commons^™

An Evaluation Of The Differential Stability Of Nucleic Acids In Biological Fluids Compromised By Degradative Treatments And Environmental Exposure, Tiffany Layne

Master of Science in Forensic Science Directed Research Projects

In an attempt to determine the boundaries of degradation in nucleic acids with increased sensitivity, and provide support for a molecular method for body fluid identification, this study used specific treatments that are commonly found in crime scenes or are present in our daily environment to degrade DNA, messenger RNA, and microRNA in bodily fluids. Recently, the forensic science field has explored the use of RNAs regarding a molecular-based method for forensic body fluid identification. Specifically, messenger RNAs and microRNAs have been researched as markers for body fluids. Messenger RNAs are multifunctional and are shorter than DNA and some are …

Consequences Of Vine Infestation: Linking Abiotic Influences And Biotic Interactions To Successional And Structural Changes In Coastal Communities, Spencer N. Bissett

Theses and Dissertations

Located at the interfaces of terrestrial and marine environments, coastal habitats are inherently vulnerable to the effects of global change. Barrier island systems in particular serve not only as protective buffers against storm events, but also as sentinel ecosystems for observation of the impacts of sea level rise, and of increasing storm frequency and intensity. In the mid-Atlantic region, shrub thickets of Morella species compose the dominant forest community. The often monospecific nature of these plant community assemblages is advantageous to ecological studies and cross-scale applications; the relatively low diversity facilitates transitions between scales. My objective was to investigate the …

Prediction Of Protein-Protein Interactions In Escherichia Coli From Experimental Data In Treponema Pallidum, Marco A. Abreu

Theses and Dissertations

Protein – Protein interactions (PPIs) are thought to be conserved between species, although this has not been systematically investigated. This problem was explored in Escherichia coli from experimental data in Treponema pallidum by predicting PPIs, focusing on protein domains of little or unknown function. The comparison of T. pallidum to a model organism such as E. coli can not only reveal additional data about T. pallidum but also reveals how E. coli is similar to this distantly related, obligate parasite. A set of novel T. pallidum interactions, enriched for proteins of unknown function, were the basis of over 23,000 predicted …

Modeling Atp-Binding Cassette G2 (Abcg2) Substrate Specificity, Raghav D. Acharya, Aurijit Sarkar, Glen E. Kellogg

UROP Posters

How well can we predict efflux by ATP-binding cassette G2?

It is estimated that there will be about 1.6 million new cases of cancer and half a million cancer deaths in the US during 2015.ATP-binding cassette (ABC) efflux transporters such as ABCG2 are overexpressed in chemotherapy-resistant cancer cells. Anticancer drugs are prone to efflux by these transporters. Being able to identify drugs that are effluxed is of great interest in drug discovery.The current arsenal of methods used to detect efflux are not easily adaptable to high throughput formats and are limited in scope, making experimental analysis an expensive prospect. Hence, …

Smoking Patterns Among Vcu Students, Jasmine Saini

UROP Posters

For some individuals, college can be a high risk time for the development of problems associated with alcohol use and other substances. The purpose of this study is to examine these initiation and use patterns as they relate to nicotine use among college students 18 years of age and older enrolled in Spit for Science: The VCU Student Survey. The Spit for Science research project evaluates how genetic and environmental factors contribute to substance use and emotional health among college students at VCU. This study uses data from the Spit for Science 2011 cohort (n=2007) to investigate smoking patterns among …

Multi-Platform Arabinoxylan Scaffolds As Potential Wound Dressing Materials, Donald C. Aduba Jr

Theses and Dissertations

Biopolymers are becoming more attractive as advanced wound dressings because of their naturally derived origin, abundance, low cost and high compatibility with the wound environment. Arabinoxylan (AX) is a class of polysaccharide polymers derived from cereal grains that are primarily used in food products and cosmetic additives. Its application as a wound dressing material has yet to be realized. In this two-pronged project, arabinoxylan ferulate (AXF) was fabricated into electrospun fibers and gel foams to be evaluated as platforms for wound dressing materials. In the first study, AXF was electrospun with varying amounts of gelatin. In the second study, AXF …

Physical And Biological Constraints On The Abundance Of Cyanobacteria In The James River Estuary, Brendan C. Trache

Theses and Dissertations

The tidal-fresh James River experiences recurring blooms of toxin-producing cyanobacteria, including Microcystis aeruginosa. However, cyanobacteria cell densities in the James are relatively low. Our purpose was to identify key factors suppressing cyanobacteria growth in the face of eutrophication. A mesocosm experiment was designed to test the effects of light, mixing and grazing on cyanobacteria abundance, with nutrients held constant. We predicted that toxic cyanobacteria would be most abundant under stagnant conditions, with enhanced light, with no bivalve grazers present. Abundances of indicator gene copies and phytoplankton counts supported this hypothesis. However, chlorophyll-a, phycocyanin, and the toxin microcystin were all …

The Role Of Srsf3 In Control Of Alternative Splicing Of Cpeb2 In Triple Negative Breast Cancer, Brian P. Griffin

Theses and Dissertations

In the presented study, we identified that SRSF3 controls the alternative splicing of CPEB2 and consequently promotes a metastatic phenotype in triple negative breast cancer (TNBC). TNBC causes thousands of deaths annually, frequently due to a lack of effective treatments and a high rate of metastasis in patients. Alternative splicing has been found to be dysregulated in numerous cancers, while splicing factors such as SRSF3 are variably expressed. In this study we performed a siRNA panel to screen potential splicing factors, then used specific siRNA to study the effect of its knockdown on cellular function. These results showed that SRSF3 …

Characterizarion Of The Regulation And Function Of Neisseria Gonorrhoeae Tonb-Dependent Transporters: Tdfg, Tdfh And Tdfj, Sophonie Jean

Theses and Dissertations

The obligate human pathogen Neisseria gonorrhoeae successfully overcomes host strategies to limit essential nutrients, termed “nutritional immunity” by expression of TonB-dependent transporters (TdTs): outer membrane receptors that facilitate nutrient transport in an energy-dependent manner. N. gonorrhoeae encodes eight TdTs, five of which facilitate utilization of iron or iron-chelates from host derived proteins including transferrin, lactoferrin and hemoglobin, in addition to siderophores from neighboring bacteria. The transferrin utilization system was previously shown to be critical for establishing infection in human males; demonstrating the possible contributions of TdTs to gonococcal pathogenesis. As such, studies describing the biological function and contribution to pathogenesis …

Xlf-Dependent Nonhomologous End Joining Of Complex Dna Double-Strand Breaks With Proximal Thymine Glycol And Screening For Xrcc4-Xlf Interaction Inhibitors, Mohammed Al Mohaini

Theses and Dissertations

DNA double-strand breaks induced by ionizing radiation are often accompanied by ancillary oxidative base damage that may prevent or delay their repair. In order to better define the features that make some DSBs repair-resistant, XLF-dependent nonhomologous end joining of blunt-ended DSB substrates having the oxidatively modified nonplanar base thymine glycol (Tg) at the first (Tg1) , second (Tg2), third (Tg3) or fifth (Tg5) positions from one 3’ terminus was examined in human whole-cell extracts. Tg at the third position had little effect on end-joining even when present on both ends of the break. However, Tg as the terminal or penultimate …

Expression And Pharmacological Modulation Of Pain-Depressed Behavior In Rats, Michael D. Leitl

Theses and Dissertations

Pain is often associated with depression of behavior and mood, and relief of pain-related depression is a common goal of treatment. This goal of this dissertation was to conduct preclinical research experiments designed to address a set of three inter-related aims that examine the expression, mechanisms and treatment of pain-related depression of Intracranial Self-Stimulation (ICSS) in rats. First, studies evaluated the hypothesis that acute acid-induced depression of ICSS was mediated by a kappa opioid receptor mediated decrease in mesolimbic dopamine release in the nucleus accumbens. Results support a role for depressed mesolimbic dopamine release in pain-related depression of ICSS; however, …

Interaction Between Atm Kinase And P53 In Determining Glioma Radiosensitivity, Syed F. Ahmad

Theses and Dissertations

Glioblastoma multiforme (GBM) is the most common primary brain tumor. Studies have shown that targeting the DNA damage response can sensitize cancer cells to DNA damaging agents. Ataxia telangiectasia mutated (ATM) is involved in signaling DNA double strand breaks. Our group has previously shown that ATM inhibitors (ATMi) sensitize GBM cells and tumors to ionizing radiation. This effect is greater when the tumor suppressor p53 is mutated.

The goals of this work include validation of a new ATM inhibitor, AZ32, and elucidation of how ATMi and p53 status interact to promote cell death after radiation. We propose that ATMi and …

Phenotypic Characterization Of Pnpase Knockdown In C. Elegans, Laura A. Lambert

Theses and Dissertations

The multifunctional exoribonuclease protein PNPase is implicated as a potential target for cancer therapy as well as causing mitochondrial disorders in humans, but there has yet to be a whole animal knockdown model created. In this study, C. elegans was used to investigate the effect of knocking down pnpt-1, the gene that encodes PNPase. It was discovered that pnpt-1 knockdown significantly extends lifespan via an increase in superoxide production similar to other known mitochondrial lifespan extension pathways. Additionally, mitochondrial networks, size and respiration are affected indication of other mitochondrial dysfunction..

PNPase is also known to transport small RNAs into the …

Investigation Into The Specification Of Nurf Recruitment To The Genome, Marissa Mack

Theses and Dissertations

The nucleosome remodeling factor (NURF) is a mutli-protein complex that plays a role in the regulation of gene expression through its ability to remodel nucleosomes. The largest subunit of this complex, Bptf (Bromodomain PHD Finger Transcription Factor) is important for many cellular processes as a transcriptional regulator and improper function results in disease or malignancy. To further understand the genome-wide recruitment of the NURF complex, the interaction partner for the N-terminal PHD finger domain of Bptf was investigated through pull down assays followed by mass spectrometry. It was determined that this domain does not recognize histones; instead it recognizes a …

Structural Determinants Of Abuse-Related Neurochemical And Behavioral Effects Of Para-Substituted Methcathinone Analogs In Rats, Julie S. Bonano

Theses and Dissertations

Methcathinone (MCAT) is the β-ketone analog of methamphetamine, and like its amphetamine analog, MCAT functions as a monoamine releaser that selectively promotes the release of dopamine (DA) and norepinephrine (NE) over serotonin (5-HT). MCAT produces amphetamine-like psychostimulant effects and is classified as a Schedule I drug of abuse by the United States Drug Enforcement Administration (DEA). Recently, synthetic MCAT analogs have emerged as designer drugs of abuse in Europe and the United States and have been marketed under deceptively benign names like “bath salts” in an attempt to evade legal restriction. These dangerous, recently emergent and novel drugs of abuse …

Cell Cycle Regulation Of Retinal Progenitors; A Role For The Nance-Horan Syndrome Protein In Retinogenesis, Paul J. Vorster

Theses and Dissertations

The Nance-Horan syndrome gene (NHS) plays a role in lens, eye and brain development. To date, the function of NHS remains unclear. Recent evidence showed that p53 isoform, Δ113p53, inhibits abnormal cell growth during organogenesis. We show that NHS is expressed in the retinas of Danio rerio and Xenopus tropicalis during key stages of retinogenesis, and that knockdown of the gene resulted in a small eye phenotype in both species. Initially, knockdown of nhsb in zebrafish had no visible defects at 24hpf. But examination of the retina at 48hpf, we see a marked difference in size compared to control embryos. …

Effects Of The Algal Toxin Microcystin On Fishes In The James River, Virginia, Maxwell D. Haase

Theses and Dissertations

With the global rise in frequency of harmful algal blooms in estuarine environments comes an increase in prevalence of toxic metabolites, such as microcystin (MC), that some of the cyanobacteria involved will produce. At high concentrations, MC may accumulate in consumer tissues and have deleterious effects on organisms; however impacts of the toxin on aquatic living resources at ecologically relevant concentrations have not been widely documented. We analyzed the effects of MC on juveniles of five fish species from the James River, Virginia to determine if MC has the potential to impede growth. Using three separate experimental approaches, it was …

Graph-Based Regularization In Machine Learning: Discovering Driver Modules In Biological Networks, Xi Gao

Theses and Dissertations

Curiosity of human nature drives us to explore the origins of what makes each of us different. From ancient legends and mythology, Mendel's law, Punnett square to modern genetic research, we carry on this old but eternal question. Thanks to technological revolution, today's scientists try to answer this question using easily measurable gene expression and other profiling data. However, the exploration can easily get lost in the data of growing volume, dimension, noise and complexity. This dissertation is aimed at developing new machine learning methods that take data from different classes as input, augment them with knowledge of feature relationships, …

Characterization Of Mechanism Of Action Of Hydrogen Sulfide (H2s) In The Regulation Of Smooth Muscle Function, Ancy D. Nalli

Theses and Dissertations

Hydrogen sulfide (H₂S) is receiving increasing interest, as much as nitric oxide (NO) and carbon monoxide have received previously, to understand its physiological functions as it meets all the criteria to define as a third gasotransmitter. Endogenous synthesis from L-cysteine via cystathionine-γ-lyase (CSE) and cystathionine-β-synthase (CBS) and the function of H₂S as an inhibitor of smooth muscle contraction in gastrointestinal tract are known. However, the loci of generation and action of H₂S, and the mechanism of inhibition of contraction are unknown. Hence, my aims in the present study are to: i) identify the expression …

Coarse Woody Debris And The Carbon Balance Of A Moderately Disturbed Forest, Amy V. Schmid

Theses and Dissertations

Landscapes are comprised of multiple ecosystems shaped by disturbances varying in severity and source. Moderate disturbance from weather, pathogens, insects, and age-related senescence, in contrast to severe disturbances that fell trees, may increase standing woody debris and alter the contribution of coarse woody debris (CWD) to total ecosystem respiration (R_E). However, woody debris dynamics are rarely examined following moderate disturbances that substantially increase standing dead wood stocks. We used an experimental manipulation of moderate disturbance in an upper Great Lakes forest to: 1) examine decadal changes in CWD stocks through a moderate disturbance; 2) quantify in situ CWD …

Heterosandwich Assay Of Nicotinic Acetylcholine Receptors, Augustine J. Pagan Iv

Theses and Dissertations

Using the technology afforded by Winschel et al., cyclen-1, a high affinity, strong complexation agent for 8-hydroxypyrene-1,3,6 trisulfonate and derivatives, a new assay has been developed for fluorescently labeling proteins of interest (POIs). Ligation of the endogenous ligand for nicotinic acetylcholine receptors (nAChRs), acetylcholine, using click chemistry afforded the triazole derivative of an alkynyl-acylcholine (compound 1) with 8-azidopyrene-1,3,6 trisulfonate (compound 2). Liposomes encapsulated with Rhodamine B were used to strengthen the initial fluorophore response of compound 2, using an anchored form of cyclen-1 complex. Using a palmitoyl tail as the lipophilic moiety for liposomal amplification, the …

Hif-Independent Responses In Hypoxia, Divya Padmanabha

Theses and Dissertations

The adaptive response to hypoxia is accompanied by widespread transcriptional changes that allow for prolonged survival in low oxygen. Many of these changes are directly regulated by the conserved hypoxia-inducible factor-1 (HIF-1) complex; however, even in its absence, many oxygen-sensitive transcripts in Caenorhabditis elegans are appropriately regulated in hypoxia. To identify mediators of these non-HIF-dependent responses, I established a hif-1 mutant reporter line that expresses GFP in hypoxia or when worms are treated with the hypoxia mimetic cobalt chloride (cobalt chloride). The reporter is selective and HIF-independent, in that it remains insensitive to a number of cellular stresses, but is …

Functional Characterization Of Rai1 In Zebrafish, Joshua S. Beach

Theses and Dissertations

Smith-Magenis Syndrome (SMS; OMIM #182290) is a multiple congenital abnormality and intellectual disability (ID) disorder caused by either an interstitial deletion of the 17p11.2 region containing the retinoic acid induced-1 (RAI1) gene or a mutation of the RAI1 gene. Individuals diagnosed with SMS typically present characteristics such as ID, self-injurious behavior, sleep disturbance, ocular and otolaryngological abnormalities, craniofacial and skeletal abnormalities, neurological and behavioral abnormalities, as well as other systemic defects and manifestations. Previous work by Vyas in 2009 showed temporal expression of rai1 in zebrafish embryos as early as 9 hpf. We hypothesize that there is maternal …

Proof-Of-Concept Of Environmental Dna Tools For Atlantic Sturgeon Management, Jameson Hinkle

Theses and Dissertations

Abstract

The Atlantic Sturgeon (Acipenser oxyrinchus oxyrinchus, Mitchell) is an anadromous species that spawns in tidal freshwater rivers from Canada to Florida. Overfishing, river sedimentation and alteration of the river bottom have decreased Atlantic Sturgeon populations, and NOAA lists the species as endangered. Ecologists sometimes find it difficult to locate individuals of a species that is rare, endangered or invasive. The need for methods less invasive that can create more resolution of cryptic species presence is necessary. Environmental DNA (eDNA) is a non-invasive means of detecting rare, endangered, or invasive species by isolating nuclear or mitochondrial DNA (mtDNA) from the …

Prothonotary Warbler Nestling Diet And Growth In Response To Variation In Aquatic And Terrestrial Food Availability, Jenna C. Dodson

Theses and Dissertations

Food supply has been suggested as the main determinant of reproductive success in birds. Riparian species can take advantage of seasonal pulses of both terrestrial and aquatic prey, though aquatic resources are often overlooked in studies of diet and reproductive ecology. This study investigates the flux of both aquatic (mayfly) and terrestrial (caterpillar) prey resources and nestling diet of the Prothonotary Warbler throughout the breeding season in two eastern Virginia sites. One site had significantly higher aquatic prey (mayfly) availability. Nestling diet was generally reflective of prey availability, and nestlings grew faster at the site with high aquatic prey availability. …

Effects Of Constant Vs. Fluctuating Temperatures On Performance And Life History Of The Herbivorous Pest Lymantria Dispar (Lepidoptera: Eribidae), Brendan E. Sostak

Theses and Dissertations

The role of temperature variation in organismal performance is understudied, but is critically important for understanding the response of biodiversity to climate change. To address this issue in herbivorous insects, I studied the direct and interactive effects of thermal regime (constant vs. fluctuating temperatures) and nutrition (dietary nitrogen) on gypsy moth (Lymantria dispar) performance under laboratory conditions. Predictions for differences between constant and fluctuating thermal conditions were derived from Jensen’s inequality, and artificial diets of differing nutritional quality were made by modifying nitrogen (casein) content. Larvae were reared in the laboratory under four temperature regimes (22°C constant, 22°C …

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 …

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) …