Life Sciences Commons^™

A Rational Design Approach To Developing Second Generation Fabry Disease Treatments, Matthew Metcalf

Doctoral Dissertations

Fabry disease is an X-linked lysosomal storage disorder that affects approximately 1 in 40,000 males in its classical form and as many as 1:4,600 in its late-onset form [1]. The disease is caused by mutations in the gene encoding α- galactosidase (α-GAL), which results in deficient levels of α-GAL activity in the lysosomes of patients [2, 3]. This lack of enzymatic activity causes macromolecular substrates to accumulate in tissues, and can result in a wide range of symptoms such as impaired renal and cardiac function [4]. The severity of disease is linked to the amount of residual …

Circadian Regulation Of The Timing Of Pre-Ovulatory Luteinizing Hormone Surge In Mice And Hamsters, Ajay Kumar

Doctoral Dissertations

Endogenous daily (circadian) oscillators ensure the proper timing of physiological and behavioral processes that are essential to health. A set of core clock genes and their protein products function in transcriptional/translational feedback loops (TTFLs) that time and coordinate vital homeostatic, cognitive, and hormonal processes. A master pacemaker in the suprachiasmatic nucleus (SCN) coordinates circadian function throughout the organism. Investigating the causal role of genetically defined cell types in the suprachiasmatic nucleus for circadian rhythms is essential to understand how an animal’s physiology is temporally organized. This thesis explores the role of molecular clocks within particular cell types in regulating physiology. …

Coffee Bioactives Regulate Lipid Metabolism In Caenorhabditis Elegans, Renalison Farias Pereira

Doctoral Dissertations

Coffee consumption is associated with reduced risk of metabolic syndrome, obesity and diabetes. Although the mechanisms of action are not yet elucidated, the effects of coffee and its bioactive components on lipid metabolism may account for the overall coffee effects on human health. Therefore, this study investigated the molecular mechanisms of coffee and its bioactive components on lipid metabolism using Caenorhabditis elegans as a model system. Green coffee bean extract (GCBE), the chlorogenic acid 5-O-caffeyolquinic acid (5-CQA), and the coffee diterpenes cafestol and kahweol reduced fat accumulation via distinct lipid metabolism pathways and/or behavior changes in C. elegans …

Engineering Stimuli-Responsive Polymeric Nanoassemblies: Rational Designs For Intracellular Delivery Of Biologics, Kingshuk Dutta

Doctoral Dissertations

Biologic drugs have gained enormous research attention in recent years as reflected by the development of multiple candidates to the clinical pipelines and an increased percentage of FDA approval. This is reasoned by the fact that biologics have been proven to deliver more predictive and promising benefits for many hard-to-cure diseases by ‘drugging the undruggable’ targets. However, the challenges associated with biologic drug development are multi-fold, viz, poor encapsulation efficacy, systemic instability, low cellular internalization and endosomal escape capability. Thus, it is essential to develop new molecular strategies that can not only address the associated drug delivery challenges, but also …

Engineered Proteins As Tools To Understand Ubiquitin Signaling, Lin Hui Chang

Doctoral Dissertations

Ubiquitin is a 76 amino acids protein that is evolutionary conserved in eukaryotes. It is an important signaling molecule in a plethora biological events, such as protein degradation, DNA damage response, and transcription. This thesis aims to develop engineered protein as a tool to study ubiquitin signaling. Through targeted mutagenesis and directed evolution, a deubiquitinase is reprogrammed into a transamidase, which lead to the generation of ubiquitinprotein conjugates with discrete ubiquitin linkages through auto-ubiquitination. These ubiquitin-protein conjugates could be used as a model substrate to profile their interaction of different ubiquitin interacting proteins. In addition, using directed evolution and deep …

The Association Of The Inflammatory Potential Of Diet With Inflammation And Depression Among U.S. Adults: Nhanes 2005-2010, Deniz Azarmanesh

Doctoral Dissertations

Depression affects 8% of adults in America. Women are twice as likely as men to experience depression. The economic burden in the U.S. is $83 billion in direct (e.g., pharmaceutical) and indirect costs (e.g., absenteeism from work). The etiology of depression includes non-modifiable (e.g., genetics) and modifiable risk factors (e.g., diet). Depression is concurrent with an increase in inflammatory biomarkers, such as c-reactive protein (CRP). Emerging research suggests that a pro-inflammatory diet may increase odds of experiencing depression. The Dietary Inflammatory Index (DII) assesses the inflammatory potential of diet. According to our narrative review of the current literature in Chapter …

Don't Delay: The Effects Of Tide Gates And Road-Stream Crossing Culverts On River Herring (Alosa Spp.) Spawning Migrations, Derrick J. Alcott

Doctoral Dissertations

River herring (Alosa spp.) are anadromous fish that spawn in freshwater rivers and lakes in North America from Florida to Nova Scotia, CA. They have been listed as a species of conservation concern due to dramatic stock declines observed in the 1970’s. Stocks have failed to show significant signs of recovery despite over a decade of harvest restrictions throughout their range. Bycatch in commercial fisheries and reduced access to spawning habitat due to anthropogenic barriers to migration, such as dams and tide gates, have been identified as major causes of the decline in herring populations. Physical barriers to …

Effects Of Tempeh Fermentation On Soy Free And Bound Phenolics: Release, Transformation, And Stimulated Production, Amadeus Driando Ahnan

Doctoral Dissertations

The fermentation of tempeh, a traditional source of protein originated in Indonesia, has been reported to enhance the health-promoting potentials of various grains, legumes, and beans. Tempeh fermentation on soybeans can modulate the bioavailability of phenolic compounds, particularly isoflavones, bioactive compounds that have been found to be protective against lung, prostrate, and colon cancers. However, the mechanism of the protective benefits was unknown. Using whole-food and in vitro models, this study addressed this research gap by investigating the effects of tempeh fermentation using various cultures on the compositions of soy free, bound, and minor phenolics, as well as their antioxidative, …

Analysis Of Titanium Dioxide Nanoparticles In Foods Using Raman Spectroscopic Techniques, Janamkumar Pandya

Doctoral Dissertations

Titanium dioxide (TiO2) and its nanoparticles (NPs) are widely used in various applications. Recently, the presence of TiO2 NPs in food and consumer products raised safety concerns to human health and the environment. The goal of this project is to explore the capability of Raman Spectroscopy in the analysis of TiO2-NPs and apply this technique for the analysis of TiO2-NPs in food and environmental samples. Two approaches, i.e. the ligand-based and the mapping-based, were evaluated. The ligand-based approach utilized the surface enhanced Raman scattering (SERS) property of the TiO2 NPs as a substrate to enhance the signal of a surface …

Controlled Membrane Remodeling By Nanospheres And Nanorods: Experiments Targeting The Design Principles For Membrane-Based Materials, Sarah Zuraw-Weston

Doctoral Dissertations

In this thesis we explore two experimental systems probing the interactions of nanoparticles with lipid bilayer membranes. Inspired by the ability of cell membranes to alter their shape in response to bound particles, we report two experimental studies: one of nanospheres the other of long, slender nano-rods binding to lipid bilayer vesicles and altering the membrane shape. Our work illuminates the role of particle geometry, particle concentration, adhesion strength and membrane tension in how membrane morphology is determined. We combine giant unilamellar vesicles with oppositely charged nanoparticles, carefully tuning adhesion strength, membrane tension and particle concentration. In the case of …

Improving Understanding Of Forest Communities And Biodiversity With Multi-Dimensional Landscape Gradients, Ben J. Padilla

Doctoral Dissertations

This dissertation was motivated by a desire to understand the effects of habitat degradation and urbanization on a single species in a single study system in western Massachusetts, the red-backed salamander (Plethodon cinereus), but along the way unexpected conceptual and methodological hurdles caused the work to grow into a multi-species, multi-region, and multi-scale endeavor. As I designed my dissertation research and began considering approaches to quantifying heterogeneity and human influence in my study landscape, I recognized inconsistencies in methods used to define and quantify landscape metrics, particularly in urban systems. To investigate further, I conducted a critical review …

Investigating The Accumulation, Sub-Organ Distribution, And Biochemical Effects Of Nanomaterials Using Mass Spectrometry, Kristen Nicole Sikora

Doctoral Dissertations

Gold nanoparticles (AuNPs) are attractive materials for use in various biomedical applications, such as therapeutic delivery, due to their unique chemical properties and modular tunability. Mass spectrometry methods, including laser desorption/ionization mass spectrometry (LDI-MS) and inductively coupled plasma mass spectrometry (ICP-MS) have been successfully used to evaluate the distribution of AuNPs in complex biological systems. As new AuNP-based materials are developed for applications in therapeutic delivery, it is essential to simultaneously develop analytical techniques that can comprehensively assess their behavior in vivo. In this dissertation, novel mass spectrometric methods have been developed and utilized to evaluate the uptake, distribution, …

Liver Kinase B1 Isoform Expression As A Regulator Of T Cell Phenotypic Stability, Heather Sherman

Doctoral Dissertations

Following activation, CD4 T cells undergo extensive metabolic and transcriptional changes to respond to external cues and differentiate into T helper (Th) cell subsets. This differentiation process was originally perceived to be terminal. However, it has been shown that T cells exhibit plasticity between subsets in the context of highly inflammatory environments. This phenomenon is especially prevalent in autoimmune conditions such as colitis and multiple sclerosis in which high levels of IL-6 promote considerable plasticity between regulatory T (Treg) cells and Th17 cells. Herein we show that Liver Kinase B1 (LKB1), a metabolic sensor that enforces energy-preserving mechanisms such as …

Micro-Physiological Models To Mimic Mucosal Barrier Complexity Of The Human Intestine In Vitro, Abhinav Sharma

Doctoral Dissertations

The mucosal barrier in the intestine is vital to maintain selective absorption of nutrients while protecting internal tissues and maintaining symbiotic relationship with luminal microbiota. This bio-barrier consists of a cellular epithelial barrier and an acellular mucus barrier. Secreted mucus regulates barrier function via in situ biochemical and biophysical interaction with luminal content that continually evolves during digestion and absorption. Increasing evidence suggests that a mucus barrier is indispensable to maintain homeostasis in the gastrointestinal tract. However, the importance of mucus barrier is largely underrated for in vitro mucosal tissue modeling. The major gap is the lack of experimental material …

Engineering Mesothelin-Binding Proteins As Targeted Cancer Diagnostics And Therapeutics, Allison Rita Sirois

Doctoral Dissertations

Cancer is a significant global health concern; and traditional therapies, including chemotherapeutics, are often simultaneously toxic yet ineffective. There is a critical need to develop targeted cancer therapeutics which specifically inhibit molecules or molecular pathways essential for tumor growth and maintenance. Furthermore, a targeted therapy is only effective when a patient's tumor expresses the molecular target; therefore, companion diagnostics, including molecular imaging agents, are a necessary counterpart of targeted therapies. Mesothelin (MSLN) is a cell surface protein overexpressed in numerous cancers, including triple-negative breast, pancreatic, ovarian, liver, and lung, with limited expression in normal tissues. Aberrant MSLN expression promotes tumor …

Soil Health Assessment For The Agroecosystems Of West Tennessee, Surendra Singh

Doctoral Dissertations

Soil health assessment is important for making informed sustainable management decisions in production systems. An established standardized method to quantify soil health is lacking and the validity of the existing methods across agroecoregions and cropping systems is not yet proven. Therefore, this study aims to evaluate the feasibility of widely discussed three soil health tests - Haney’s Soil Health Test (HSHT), Comprehensive Assessment of Soil Health (CASH), and Alabama Soil Health Index (ASHI) to assess soil health in diverse cropping systems of Tennessee. Since these approaches were originally developed for specific agroecoregions, we hypothesized that these tests are not sensitive …

Costs Of Protected Areas In The United States, Diane Le Bouille

Doctoral Dissertations

Protected areas, or land owned in fee by agencies and non-profits to further conservation goals, have traditionally been the go-to choice for conservation interests. The UN Environment World Conservation Monitoring Centre estimates that, currently, close to 15% of all terrestrial and inland water areas are protected. This figure falls short of the Aichi Biodiversity Target of 17% in 2020, that was added to the Convention on Biological Diversity by its 196 signatories in 2010. But as the Convention prepares to set new post-2020 targets, this percentage is expected to keep increasing. Although acquiring a parcel of land is only one …

Computer Simulations Of Biological Systems: From Protein Dynamics To Drug Discovery, Rupesh Agarwal

Doctoral Dissertations

Computational biophysics methods such as molecular dynamics (MD) simulations are often used in combination with experimental techniques like neutron scattering, NMR, and FTIR to explore protein conformational landscapes. With the improvements in experimental techniques, there is also a need to continually optimize the MD forcefield parameters to make precise predictions that match experimental results. To complement many of these experiments, an accurate model of deuteration is frequently required, but has been elusive. In our work, we developed a novel method to capture isotope effects in classical MD simulations by re-parameterization of the bonded terms of the CHARMM forcefield using quantum …

Efficacy Of A Cycling Intervention With Pedal Reaction Force Augmented Feedback On Reducing Inter-Limb Asymmetries In Patients With Unilateral Total Knee Arthroplasty, Erik T. Hummer

Doctoral Dissertations

Fifteen patients with unilateral total knee arthroplasty (TKA) performed cycling at two workates (80 W and 100 W) and two walking conditions (preferred and fast speeds). Ten of these patients of TKA also participated in a short-term cycling intervention paired with visual augmented feedback of vertical pedal reaction forces for six sessions over two-three weeks. These ten patients of TKA participated in a 2^nd post-training testing session. Study One compared the knee joint biomechanics for all fifteen participants during stationary cycling to ascertain if any biomechanical asymmetries may be present. The replaced limbs displayed significantly lower peak knee extension …

Prevalence Of Rats And Rodent Borne Pathogens Across Post-Katrina New Orleans, Bruno Marco Ghersi

Doctoral Dissertations

Disasters are happening at an increasingly higher rate and intensity a trend that is expected to continue as more humans migrate to coastal urban areas. Disasters, and as importantly, disaster recovery can affect how native and pest populations will recover. My aim was to improve understanding of disease risk by evaluating the socioecological conditions that have shaped commensal rat recovery and distribution, as well as the pathogens they carry, across New Orleans after Hurricane Katrina. I first estimated relative abundance and distribution of commensal rats from rodent trapping conducted between mid-2014 and early-2017 across 96 sites in 10 areas of …

Extracting Detailed Metabolic Information And Connections From Mammalian Gut Microbiomes Via Metaproteomics, Jose A. Blakeley-Ruiz

Doctoral Dissertations

A diverse community of bacteria populates the mammalian gastrointestinal tract. These populations exist in a balance with the host assisting with key functions, particularly metabolism of intractable fibers and immune modulation. Disruption of this balance can lead to diseases such as infection, inflammatory bowel syndrome, and obesity. Common symptoms include chronic pain, chronic inflammation, and altered metabolism. Several taxonomic classifications of bacteria have been associated with these diseases, but Recent studies have indicated that these finding are not always statistically valid. An explanation for this is that microbial communities between individuals and even across time can vary substantially even when …

More Than The Sum Of Their Parts: Building A Framework For Understanding Host-Microbe Interactions In Medicago Sativa, Katherine Mackenzie Moccia

Doctoral Dissertations

This dissertation seeks to understand plant-microbe interactions in the agriculturally relevant plant Medicago sativa from three distinct vantage points within microbiology. Within the plant microbiome, we examine how primer usage and the application of peptide nucleic acids impacts 16S and 18S rRNA gene sequencing. In doing so, we design a novel peptide nucleic acid, PNA, and test its impact using multiple primers and sequencing protocols. Once microbial sequencing methodology is established, we generate a synthetic consortium of bacterial isolates from M. sativa leaves and modulate nitrogen levels to better understand microbial structure. Drop out communities, where we remove one member …

Approaches To Studying Bacterial Biofilms In The Bioeconomy With Nanofabrication Techniques And Engineered Platforms., Michelle Caroline Halsted

Doctoral Dissertations

Studies that estimate more than 90% of bacteria subsist in a biofilm state to survive environmental stressors. These biofilms persist on man-made and natural surfaces, and examples of the rich biofilm diversity extends from the roots of bioenergy crops to electroactive biofilms in bioelectrochemical reactors. Efforts to optimize microbial systems in the bioeconomy will benefit from an improved fundamental understanding of bacterial biofilms. An understanding of these microbial systems shows promise to increase crop yields with precision agriculture (e.g. biosynthetic fertilizer, microbial pesticides, and soil remediation) and increase commodity production yields in bioreactors. Yet conventional laboratory methods investigate these micron-scale …

Genetic And Epigenetic Control Of Soybean Agronomic Traits, Daniel Niyikiza

Doctoral Dissertations

Greater soybean productivity depends on the genetic improvement of yield components, epigenetic effects and the interactions with surrounding environment. We explored the possibility of soybean improvement by conventional biparental crossing of soybean lines, differing by seed quality, yield, and resistance to a range of pathogens, including soybean cyst nematode (SCN). We investigated the role of Resistance to Heterodera glycines (Rhg) 1 and 4 loci in soybean resistance to SCN, race 2 and 3. Further, we evaluated the adaptability of temperate-origin soybean lines in tropical environments of Rwanda. Lastly, we examined gene expression, RNA splicing, DNA methylation and their interactions in …

A Review Of Horticultural Therapy And Horticultural Therapy Education In The United States: Addressing Challenges And Opportunities, Derrick R. Stowell

Doctoral Dissertations

The Horticultural Therapy (HT) profession has been formally organized in the United States since 1973. Despite the long history of horticulture being used as therapy, the profession of HT in the United States has not advanced as quickly as other allied healthcare professions. This study will review the current challenges and opportunities for HT and HT education in the United States.

A qualitative study of the status of the profession was conducted to determine what challenges and opportunities the profession of HT currently has and how the profession can meet those challenges and opportunities. The study sample was recruited from …

Investigating Transmembrane-Lipid Interactions Of Epha2 And Ph Responsive Peptides, Katherine M. Stefanski

Doctoral Dissertations

Single-pass membrane receptor signaling plays vital roles in human development and maintaining homeostasis. These membrane receptors can also have causative functions in several diseases including cancer. Much is known about the structure and signaling outcomes of these receptors but the mechanistic details of how they pass an extracellular signal across the membrane and into cytoplasm via the transmembrane (TM) domain is unclear. It is further unknown how or if interactions with membrane lipids facilitate and/or regulate these events. Here we use the TYPE7 peptide to target the TM region of a receptor tyrosine kinase, EphA2. EphA2 engages in both tumorigenic …

Probing Structure, Function And Dynamics In Bacterial Primary And Secondary Transporter-Associated Binding Proteins, Shantanu Shukla

Doctoral Dissertations

Substrate binding proteins (SBPs) are ubiquitous in all life forms and have evolved to perform diverse physiological functions, such as in membrane transport, gene regulation, neurotransmission, and quorum sensing. It is quite astounding to observe such functional diversity among the SBPs even when they are restricted by their fold space. Therefore, the SBPs are an excellent set of proteins that can reveal how proteins evolution novel function in a structurally conserved/constrained fold. This study attempts to understand the phenomenon of affinity and specificity evolution in SBPs by combining a set of biochemical, biophysical, and structural studies on the SBPs involved …

Soil Moisture Sensitivity Of Microbial Processing Of Soil Organic Carbon, Shikha Singh

Doctoral Dissertations

Soil organic carbon (SOC) is the largest terrestrial C pool and understanding SOC decomposition in response to environmental factors is critical for accurate predictions of climatic change. Soil moisture is one of the most important, yet less explored, environmental factors controlling soil microbial respiration. The relationship between soil moisture and respiration also varies with soil texture. Currently, it is difficult to predict feedbacks to climatic changes from changes in soil moisture, as most earth system models lack site-specific, experimentally-derived parameters to represent soil moisture-texture-respiration relationships. The overarching goals of this dissertation are to gain a fundamental understanding of the interactive …

Root Stage Distributions And Their Importance In Plant-Soil Feedback Models, Tyler Poppenwimer

Doctoral Dissertations

Roots are fundamental to PSFs, being a key mediator of these feedbacks by interacting with and affecting the soil environment and soil microbial communities. However, most PSF models aggregate roots into a homogeneous component or only implicitly simulate roots via functions. Roots are not homogeneous and root traits (nutrient and water uptake, turnover rate, respiration rate, mycorrhizal colonization, etc.) vary with age, branch order, and diameter. Trait differences among a plant’s roots lead to variation in root function and roots can be disaggregated according to their function. The impact on plant growth and resource cycling of changes in the distribution …

A High Resolution Study Of Long-Term Vertebrate Decomposition In Human And Animal Model Systems, Lois S. Taylor

Doctoral Dissertations

The effects of vertebrate decomposition are wide-ranging across multiple foodwebs, and have been shown to persist in the environment, however there is a lack of systematic assessment of these changes over long periods of time or in sufficiently high resolution as to resolve seasonal flux patterns. The ultimate aim of this body of research was to explore nematode systematics in decomposition environments, culminating in a pair of long-term human decomposition seasonal trials, in high resolution, with the specific intent of integrating the fields of soil chemistry, microbial ecology, and nematology in order to assess the relationships of cross-disciplinary impacts. Of …