Life Sciences Commons^™

An Identification Of Tgf-B'S Role In Ischemic Cholangiopathy, Courtney Chmielewski

Theses and Dissertations

Organ transplants are a necessary intervention for many diseases that result in end stage organ failure. The donation pool cannot match the demands of the transplant list, so expanding the pool to include donation after cardiac death (DCD) is desired. However, there are increased odds of graft failure and ischemic cholangiopathy leading to inferior outcomes when DCD livers are used. Ischemic cholangiopathy consists of multiple diffuse strictures and fibrosis of the bile ducts leading to a loss of epithelialization and fibrosis. The cellular mechanism is proposed to be epithelial-to-mesenchymal transition (EMT). TGF-�� is seen as a key initiator of EMT, …

Senescent Trees Stabilize Aboveground Wood Net Primary Production Immediately After Disturbance, Maxim S. Grigri

Theses and Dissertations

In the United States, forests sequester 17% of national carbon (C) emissions annually (UGCRP, 2018), however shifting forest disturbances threaten the stability of this essential C sink. Unlike the high severity, stand-replacing disturbances that were widespread a century ago, today’s eastern temperate forests experience frequent low-to-moderate severity disturbances from invasive pests and pathogens with mixed effects on net primary production (NPP). Carbon cycling stability after disturbance has been reported, however, the mechanisms underlying immediate NPP stability or decline are not well understood. Through weekly measurements of production in a landscape scale experiment, we show that the sustained growth of senescent …

Role Of Clic4 And The Synaptic Transcriptome In The Behavioral And Molecular Neurobiology Of Ethanol, Rory M. Weston

Theses and Dissertations

Alcohol use disorder (AUD) is a prevalent neuropsychiatric disease with profound health, social, and economic consequences. With an estimated 50% heritability, identifying genes that engender risk and contribute to the underlying neurobiological mechanisms represents an important first step in developing effective treatments. Gene expression studies are an important source of candidate genes for studying AUD, providing windows into the molecular machinery engaged by the brain in response to ethanol. Published studies have identified chloride intracellular channel 4 (Clic4) as an ethanol-regulated gene in brain capable of modulating sensitivity to sedation in multiple species. The functions of Clic4 are …

Non-Destructive, Rapid Differentiation Of Cell Types Relevant To Sexual Assault Investigations Utilizing Morphological And Autofluorescence Signatures, Taylor R. Moldenhauer

Theses and Dissertations

Current methods for confirming the presence of spermatozoa in sexual assault samples can be time-consuming and often lack sensitivity; however, this remains the most definitive test for the presence of semen. Additionally, male DNA can be deposited without the presence of intact sperm as may be the case with seminal fluid from vasectomized individuals or sexual activity where seminal fluid is not recovered (e.g., perpetrator wears a condom, penetration without ejaculation, etc.). The ability to detect bodily fluids, as well as quantify their presence in a sample, could aid in forensic DNA analysis by limiting the amount of time performing …

The Role Of Ca2+/Calmodulin Dependent Protein Kinase Ii, Camk-Ii, In Kidney Morphogenesis, Sarah R. Ingram

Theses and Dissertations

Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common heritable diseases in the world, characterized by the development of large fluid filled cyst. Treatments for this disease are limited due to a lack of understanding of disease pathogenesis. Ca²⁺/calmodulin dependent protein kinase II (CaMK-II) is necessary for kidney morphogenesis in zebrafish as well as being a downstream effector of pkd2, one of the genes most commonly mutated in ADPKD patients. The roles of CaMK-II during zebrafish kidney development include regulation of kidney cell migration as well as cloacal cilia stability. The influence of CaMK-II …

Validation Of Ninein As An Ethanol-Related Quantitative Trait Gene: Reassessment, Design, And Functional Validation Of Reference Genes For Qpcr Analysis Of Brain Tissue In Mice, Jessica L. Jurmain

Theses and Dissertations

The increasing use of quantitative real-time polymerase chain reaction (qPCR) as a method for quantifying gene expression has led to an increased demand for standardization of data analysis methods to ensure accurate reporting and robust, reproducible results. The exponential nature of qPCR amplification results in the potential magnification of what are usually very small sources of error. Relative gene expression calculations circumvent this issue by normalizing target gene expression data to within-sample expression of a previously validated, stably expressed reference gene or genes. Multiple studies discussed herein have found that qPCR data are more reliable and reproducible when multiple reference …

Focal Augmentation Of Somatostatin Interneuron Function And Subsequent Circuit Effects In Developmentally Malformed, Epileptogenic Cortex, Nicole Ekanem

Theses and Dissertations

Drug-resistant epilepsy (DRE) is a common clinical sequela of developmental cortical malformations such as polymicrogyria. Unfortunately, much remains unknown about the aberrant GABA-mediated circuit alterations that underlie DRE's onset and persistence in this context. To address this knowledge gap, we utilized the transcranial freeze lesion model in optogenetic mice lines (Somatostatin (SST)-Cre or Parvalbumin (PV)-Cre x floxed channelrhodopsin-2) to dissect features of the SST, PV, and pyramidal neuron microcircuit that are potentially associated with DRE. Investigations took place within developmental microgyria’s known pathological substrate, the adjoined and epileptogenic paramicrogyral region (PMR). As well, microcircuit relationships within the previously unexplored range …

Epigenetic Alterations At Synaptic Plasticity Genes In A Genetically Heterogeneous Rat Model Of Neuropsychiatric Disorders, Doan M. On

Theses and Dissertations

Sensorimotor gating impairments are observed across a range of neuropsychiatric conditions. The prepulse inhibition of the acoustic startle response (PPI) is a validated measure of sensorimotor gating. Genetic and pharmacological manipulations in rodents have shown PPI is regulated by specific brain monoaminergic systems. Using genetically heterogeneous NIH-HS rats, we stratified individuals by %PPI. In low PPI animals, we observed elevated mRNA levels of certain neurotransmitter receptors, including metabotropic glutamate receptor Grm2, dopamine receptors Drd1 and Drd2, serotonin receptors Htr1a and Htr2a, and scaffolding protein Homer1, in the frontal cortex (FC) and striatum (STR). We found Drd2 …

Synaptic Plasticity In A Visual Cortical Region Induced By Early-Deafness, John M. Kay

Theses and Dissertations

When organisms learn and adapt to their environment or lose a sensory modality, neurons in the brain undergo a cellular process called ‘plasticity.’ This thesis explores the loss of a non- visual system (early deafness) and how it can affect visual plasticity. To examine this question, Golgi-stained cortical neurons were studied from the visual region PLLS from early-deaf cats and their hearing controls. Dendritic spine density and dendritic spine diameter are well-known indicators of synaptic plasticity and these neuronal features were measured using light microscopic techniques and Neurolucida. Within the visual PLLS, the mean spine density for the deaf cats …

The Impact Of Aging And Mechanical Injury On Alveolar Epithelial And Macrophage Responses In Acute Lung Injury And Inflammation, Michael S. Valentine

Theses and Dissertations

Patients with severe lung pathologies, such as Acute Respiratory Distress Syndrome (ARDS), often require mechanical ventilation as a clinical intervention; however, this procedure frequently exacerbates the original pulmonary issue and produces an exaggerated inflammatory response that potentially leads to sepsis, multisystem organ failure, and mortality. This acute lung injury (ALI) condition has been termed Ventilator-Induced Lung Injury (VILI). Alveolar overdistension, cyclic atelectasis, and biotrauma are the primary injury mechanisms in VILI that lead to the loss of alveolar barrier integrity and pulmonary inflammation. Stress and strains during mechanical ventilation are believed to initiate alveolar epithelial mechanotransduction signaling mechanisms that contribute …

Interspecific Gene Flow Potentiates Adaptive Evolution In A Hybrid Zone Formed Between Pinus Strobiformis And Pinus Flexilis, Mitra Menon

Theses and Dissertations

Species range margins are often characterised by high degrees of habitat fragmentation resulting in low genetic diversity and higher gene flow from populations at the core of the species range. Interspecific gene flow from a closely related species with abutting range margins can increase standing genetic diversity and generate novel allelic combinations thereby alleviating limits to adaptive evolution in range margin populations. Hybridization driven interspecific gene flow has played a key role in the demographic history of several conifer due to their life history characteristics such as weak crossability barriers and long generation times. Nevertheless, demonstrating whether introgression is adaptive …

Model-Based In-Vitro Pk/Pd Profiling Of Novel Synthetic Allosteric Effectors Of Hemoglobin (Aeh) As Potential Sickle Cell Disease (Scd) Therapeutics, Xiaomeng Xu

Theses and Dissertations

Introduction: Allosteric effectors of hemoglobin (AEH) represent a class of synthetic aromatic aldehydes that transiently form covalent interactions (Schiff-base) with hemoglobin (Hb) to form Hb-AEH adduct, preventing the HbS polymerization and sickling of red blood cells (RBC). The overall objective of this research was to aid in the optimization of novel AEH by understanding their target-site disposition of AEH in relevant biological matrices, e.g., HbA solution, whole blood (WB) and human liver cytosol (HLC), a surrogate of aldehyde dehydrogenase (ALDH)-mediated oxidative metabolism.

Methods: A “universal” HPLC-UV/Vis assay method was developed for the quantitation of HbA-AEH adduct for chemically …

The Human Intermediate Prolactin Receptor: A Breast Cancer Proto-Oncogene, Jacqueline M. Grible

Theses and Dissertations

The hormone prolactin (PRL) and its receptor (hPRLr) are significantly involved in breast cancer pathogenesis. The intermediate hPRLr (hPRLrI) is an alternatively-spliced isoform, capable of stimulating cellular viability and proliferation. An analogous truncated mouse PRLr (mPRLr) was recently found to be oncogenic when co-expressed with wild-type mPRLr. hPRLrI co-expression with full-length hPRLr (hPRLrL) in MCF10AT cells resulted in robust in vivo and in vitro transformation, while hPRLrI knock-down in MCF7 cells significantly decreased in vitro malignant potential. hPRLrL+I heterodimers displayed greater stability than hPRLrL homodimers, and while being capable of activating Jak2, Ras, and MAPK, they were unable to induce …

Stimulated Growth Response To Sand Burial Of A Coastal Shrub, D. Nicole Keller

Theses and Dissertations

Drivers of vegetation zonation on barrier islands are complex and interconnected. Sand burial is a strong driver in dynamic coastal systems, especially in the foredune community. However, it is not well understood how burial impacts the interdunal swales communities and it is especially difficult to separate the effects of burial from salinity. Climate change is altering the frequency of overwash events as well as expanding the range of the native shrub, Morella cerifera, on the Virginia barrier islands. To accurately forecast island response to climate change it is important to understand how the shrub responds to sand burial. Juvenile …

Ndrg1 And Myelin-Related Disease: Alcoholism And Chemotherapy-Induced Neuropathy, Guy Harris

Theses and Dissertations

Alcohol use disorder (AUD) is a prevalent neuropsychiatric disease with profound health, social, and economic consequences. With an estimated 50% heritability, identifying genes that engender risk and contribute to the underlying neurobiological mechanisms represents an important first step in developing effective treatments. Gene expression studies are an important source of candidate genes for studying AUD, providing windows into the molecular machinery engaged by the brain in response to ethanol. Our laboratory has implicated N-myc down-regulated gene 1 (Ndrg1) as a potential candidate gene that modulates ethanol-induced changes in myelin-related gene expression and acute sensitivity to ethanol. Analysis of …

Characterization Of The Dyrk1a Protein-Protein Interaction Network, Varsha Ananthapadmanabhan

Theses and Dissertations

Human Dual specificity tyrosine (Y)-Regulated Kinase 1A (DYRK1A) is a protein kinase encoded by a dosage-dependent gene. An extra copy of DYRK1A contributes to Down syndrome (DS) pathogenesis while loss of one allele causes severe mental retardation and autism. DYRK1A is involved in phosphorylation of several proteins that regulate cell cycle control and tumor suppression. However, the function and regulation of this kinase is not well understood and current knowledge does not fully explain dosage-dependent function of this important kinase. Our previous proteomic studies identified several novel DYRK1A interacting proteins including RNF169, FAM117B, TROAP, LZTS1, LZTS2 and DCAF7. In this …

Characterization Of The Tsc/Dyrk1a Interaction, Supriya Joshi

Theses and Dissertations

The Tuberous sclerosis complex (TSC) includes TSC1, TSC2 and the TBC1D7 subunits that together function as a principal inhibitor of the mTOR protein kinase complex 1 (mTORC1). mTORC1 is a master regulator of cell growth and proliferation that responds to signaling cues such as growth factors and nutrient availability. Proteomic studies in our lab revealed an interaction between the TSC subunits and DYRK1A, a ubiquitous protein kinase encoded by a gene located in the Down syndrome (DS) region on human chr21. In this study, we sought to validate the interaction of the TSC components with DYRK1A and to determine the …

Higher Tensile Forces Across Cellular Junctions And An Intact Nuclear Linc Complex Is Required For Epithelial Function And Stability, Fnu Vani Narayanan

Theses and Dissertations

Recent advances in three-dimensional (3D) cell culture systems have provided key insights into the understanding of biochemical and physiological states of native tissue. A significant progress in the field of mechanobiology involves measuring cellular traction forces in a more native 3D environment. However, the effects of mechanical forces exerted across cellular junctions and the nuclear LINC complex, in an organized 3D system has not been investigated thus far. Epithelial cells spontaneously form acini (also known as cysts or spheroids) with a single, fluid-filled central lumen, when grown in 3D matrices. The size of the lumen is dependent on apical secretion …

Mathematical Models Of Cellular Signaling And Supramolecular Self-Assembly, Pratip Rana

Theses and Dissertations

Synthetic biologists endeavor to predict how the increasing complexity of multi-step signaling cascades impacts the fidelity of molecular signaling, whereby cellular state information is often transmitted with proteins diffusing by a pseudo-one-dimensional stochastic process. We address this problem by using a one-dimensional drift-diffusion model to derive an approximate lower bound on the degree of facilitation needed to achieve single-bit informational efficiency in signaling cascades as a function of their length. We find that a universal curve of the Shannon-Hartley form describes the information transmitted by a signaling chain of arbitrary length and depends upon only a small number of physically …

The Influence Of Breeding Density On Female Aggression, Parental Care, And Ornamentation In A Secondary-Cavity Nesting Warbler, Elsa B. Chen

Theses and Dissertations

Tradeoffs between individual survival and reproductive success associated with aggressive behaviors are a driving force of evolution, but these tradeoffs are often overlooked for aggressive conspecific interactions between females. For avian males, it is well documented that more aggressive individuals tend to provide less parental care. In the few studies that address this in females, the tradeoffs seem to be more context-dependent, varying due to factors such as predation pressure and habitat quality. The relationship between female ornamentation and aggression is similarly understudied, but evidence suggests that both aggression and ornamentation are important traits involved in social selection – the …

Plant-Pollinator Associations In An Eastern Serpentine Savannah And The Effects Of Overbrowsing, Allyson Richins

Theses and Dissertations

Chapter 1: Native plant response to deer overbrowsing in a serpentine savannah

Plants are particularly vulnerable to physical disturbance in low productivity areas, due to a high cost of replacing lost plant tissue. In the eastern United States, serpentine grasslands are fragmented ecosystems with high concentrations of rare endemic plant species, low concentrations of soil nutrients, and uncontrolled deer overpopulation. This study assessed functional responses of native angiosperms in a rare eastern serpentine savannah to selective deer browsing. Plant count, flower count, floral area, vegetative area, and plant height of 10 serpentine plant species were compared inside and outside of …

Mef2-Bound Genes May Influence Ethanol Sedation In Drosophila Melanogaster, Katlyn M. Myers

Theses and Dissertations

Alcohol Use Disorder (AUD) is a global health issue that affects millions of people every year. This disorder has serious negative mental and physical consequences. Currently, treatment options for this disorder are largely limited to psychological therapy, with very few medications available to treat it. Being able to identify the environmental and genetic components that influence AUD can help improve diagnosis and treatment options. Previous studies in humans have shown a link between initial sensitivity and risk for alcohol abuse. Our laboratory uses Drosophila melanogaster as a model to study the genetic and environmental components of alcohol-related behaviors. Previous lab …

Spag17 Is Important For Protein Trafficking In Mammalian Spermiogenesis, Virali Bhagat

Theses and Dissertations

Spermiogenesis is the process through which undifferentiated germ cells develop into mature spermatozoa. While spermiogenesis is a very well-regulated process, the protein-protein interactions regulating it remain poorly understood. A knockout (KO) mouse for the ciliary protein SPAG17 was generated by our lab. Loss of SPAG17 has been shown to disrupt the transport of proteins important for acrosome biogenesis and manchette functions. With this information, we hypothesized that SPAG17 plays an essential role in protein trafficking during mammalian spermiogenesis. To further investigate this, immunofluorescence (IF) studies were performed in germ cells collected from both WT and SPAG17 KO mice to visualize …

The Gamma-Globin Repressors Klf1, Bcl11a, And Mbd2: Knowledge Gaps In The Literature, Lauren Nufer

Theses and Dissertations

β-hemoglobinopathies affect millions of people around the world. Research into treatments for these conditions has focused on methods to increase γ-globin expression, because increased levels of γ-globin ameliorate or reduce the severity of symptoms. As more and more studies have been done, a few proteins have emerged as having crucial roles in γ-globin repression and have been established as key genes to study. These are Krüppel-like factor 1 (KLF1), B cell CLL/lymphoma 11A (BCL11A), and methyl binding domain 2 (MBD2). The roles of these proteins in the switch from fetal to adult hemoglobin and in repressing γ-globin expression have been …

The Role Of Manganese In Streptococcus Sanguinis, Tanya M. Puccio

Theses and Dissertations

Streptococcus sanguinis is primarily associated with oral health as a commensal bacterium. As an opportunistic pathogen, S. sanguinis is capable of colonizing heart valve vegetations, leading to the disease infective endocarditis. Previous studies from our lab have identified the high-affinity manganese transporter SsaACB as important for endocarditis virulence. The impact that manganese depletion has on S. sanguinis had never been evaluated and a secondary manganese transporter has not been identified. Thus, we employed the use of a fermentor to control large-scale growth over time and depleted manganese in an ΔssaACB mutant using a metal chelator, EDTA. The changes in …

Investigation Of Molecular Mechanisms Of Liver Preservation Injury: A Complication Preceding Organ Transplantation, Ria C. Fyffe-Freil

Theses and Dissertations

Of the over 108,000 American awaiting a life-saving organ transplant today, over 12,000 (11%) of those need a new liver (OPTN, 2020). Last year, only 35% of patients on the waiting list for an organ were transplanted. Improving the quality of marginal organs by preventing or reversing preservation injury could vastly increase the number of transplants performed. Washout of circulating blood during liver procurement with cold University of Wisconsin solution flushes out any endogenous pro-survival signaling molecules. We investigated lysophospholipid (LPL) surface receptors (G-protein coupled receptors for lysophosphatidic acid (LPA) and sphingosine- 1-phosphate (S1P)) and their role in protecting hepatocytes …

Taxa Specific Responses To Flooding Shape Patterns Of Abundance In A River Rock Pool Metacommunity, Charles R. Stunkle

Theses and Dissertations

Connectivity and habitat area are important landscape characteristics that drive patterns of abundance and diversity across scales. However, responses to connectivity and patch size are dependent on species traits. Riverine landscapes are highly dynamic both spatially and temporally with hydrologic connectivity being a major driver of abundance and diversity. Here we modeled densities of the Virginia river snail and skimmer dragonfly nymphys, two taxa with differences in their dispersal abilities and life histories, as a function of flooding, patch area, and season in over 300 riverine rock pools. We found key differences in how each taxon responded to these predictors …

Transcriptomic Profiling Of Postmortem Prefrontal Cortex And Nucleus Accumbens From Chronic Alcohol Abusers., Eric S. Vornholt

Theses and Dissertations

Alcohol use disorder (AUD) is a debilitating psychiatric illness that develops from a combination of genetic and environmental factors. While it is well documented that AUD is heritable, the shift from recreational alcohol use to abuse/dependence is poorly understood. In this dissertation, using postmortem brain tissue from individuals with alcohol dependence (AD), we profiled the genome-wide expression of circular RNA (circRNA), microRNA (miRNA), and messenger RNA (mRNA) to better understand the impact of gene expression on the development of AUD. To achieve this, we performed two independent studies that explore transcriptome differences between AD cases and controls. The first of …

Effects Of Gestational Ozone Exposure On Privileged Placental And Brain Barrier Integrity, Alexander I. Hamm

Theses and Dissertations

Ambient outdoor ozone, a common of component of photochemical smog and urban air pollution, is linked to various neurological and vascular pathologies. Its immediate reaction with lung surfactant after inhalation results in complete reactivity of the gas, with no active ozone passing into circulation. This indicates the presence of secondary and tertiary mediators in ozone-related systemic pathologies after pulmonary insult. In vasculature, ozone exposure is associated with an acute hypertensive phenotype apparent at least 24 hours after dose, such as experienced on a hot summer afternoon in a large metropolitan area like Los Angeles or Mexico City. However, the effects …

Assessing The Feasibility Of Freshwater Mussel Restoration In Urban Streams., John-Reid Ryan

Theses and Dissertations

Abstract

The main objective of this study was to determine whether introduced freshwater mussels (Alewife floater, Utterbackiana implicata) can survive and grow in urban streams in the James River watershed. A secondary objective was to assess differences in U. implicata survival and growth of in the context of differing water quality and food resource conditions among three urban sites and three rural sites. Results from this study show large differences in growth and survivorship of mussels across sites. Higher survivorship was observed among mussels stocked into rural streams (35% and 44%) in comparison to urban streams (3%, 6% and 14%). …