Life Sciences Commons^™

Functional And Structural Characterization Of The Mevalonate Diphosphate Decarboxylase And The Isopentenyl Diphosphate Isomerase From Enterococcus Faecalis, Chun-Liang Chen

Open Access Dissertations

Enterococcus faecalis causes a diverse range of nosocomial infections (in wounds, the gastrointestinal tract, the blood stream and the endocardium), and multidrug-resistant strains have become a serious issue across countries. Vancomycin, a FDA-approved drug for the disruption of the bacterial cell wall biosynthesis, has been utilized to treat infectious diseases caused by Enterococci; however, the prevalence of vancomycin-resistant enterococci (VRE) threatens communities all over the world. We aim at developing novel therapeutic strategies to control bacterial growth of Enterococci, and we focus on targeting two essential enzymes involved in poly-isoprenoid biosynthesis in Enterococcus faecalis; one is the mevalonate diphosphate decarboxylase …

Ros Regulation Of Axonal Mitochondrial Transport, Pin-Chao Liao

Open Access Dissertations

Mitochondria perform critical functions including aerobic ATP production and calcium (Ca2+) homeostasis, but are also a major source of reactive oxygen species (ROS) production. To maintain cellular function and survival in neurons, mitochondria are transported along axons, and accumulate in regions with high demand for their functions. Oxidative stress and abnormal mitochondrial axonal transport are associated with neurodegenerative disorders. However, we know little about the connection between these two. Using primaryDrosophila neuronal cell culture and the third instar larval nervous system as in vitro and in vivo models, respectively, we studied mitochondrial transport under oxidative stress conditions. In vitro …

The Role Of Seed Attributes In Eastern Gray Squirrel Foraging, Mekala Sundaram

Open Access Dissertations

Seed attributes are important predictors of rodent foraging behaviors. I examined the role of seed attributes in eastern gray squirrel (Sciurus carolinensis) foraging behavior from an evolutionary, economic, ecological and biochemical perspective. From an evolutionary perspective (chapter 2), I found that squirrel foraging behaviors are influenced by a combination of phylogenetically conserved and evolutionarily labile seed traits, which supports a diffuse coevolutionary relationship between hardwood trees and squirrels and provides indirect evidence supporting the Janzen-Connell and handling time hypotheses. From an economic perspective (chapter 3), I found that eastern gray squirrels are homogenous with respect to their preferences for seed …

The Relationship Between Protein And Phosphorus Digestion And Retention In Growing Pigs And Broiler Chickens, Pengcheng Xue

Open Access Dissertations

Xue, Pengcheng. Ph.D., Purdue University, December 2016. The Relationship between Protein and Phosphorus Digestion and Retention in Growing Pigs and Broiler Chickens. Major Professor: Dr. Layi Adeola. The objective of this study was to investigate the relationship between protein and P digestion and retention in growing pigs and broiler chickens. The methodology of determining the digestibility of AA and P and the effect of dietary N and P on the digestion and retention of these two nutrients were investigated.

An experiment was conducted in growing pigs to investigate the additivity of AID or SID of CP and AA in mixed …

Synthesis And Performance Of Novel Supramolecular Tools For Single-Particle Cryogenic Electron Microscopy And Drug And Gene Delivery, Kyle J. Wright

Open Access Dissertations

High-resolution biomacromolecular structure elucidation is fundamentally important to structure-based drug design and basic research into complex biochemical processes. Cryo-EM is an emerging alternative to XRD and NMR that is complementary in many ways relative to XRD and NMR. Materials approaches to cryo-EM are anticipated to greatly facilitate the cryo-EM process, allowing progress toward a more high-throughput application of cryo-EM to address challenges in structural biology. ^ Various affinity-based approaches inspired by approaches previously introduced for 2D crystallization were developed for facilitation of cryo-EM. A library of affinity lipopolymer constructs were synthesized consisting of lipopolymers of various PEG molecular weights conjugated …

Graphical Methods In Rna Structure Matching, Jiajie Huang

Open Access Dissertations

Eukaryotic genomes are pervasively transcribed; almost every base can be found in an RNA transcript. This is a surprising observation since most of the genome does not encode proteins. This RNA must serve an important regulatory function – important because producing non-coding RNA is an energy intensive process, and in the absence of strong selection one would expect it to disappear.

RNA families with common functions have specifically conserved structural motifs, which are directly related to the functional roles of RNA in catalysis and regulation. Because the conserved structures depend on base-pairing, similar RNA structures may have little or no …

Understanding Plant Response To Stress Using Gene Model Quality Evaluation And Transcriptome Analysis, Karthik Ramaswamy Padmanabhan

Open Access Dissertations

The overall aim of the project was to understand how plants reacted to environmental stress and evolved to overcome it. The land plants that we see today evolved from a green algal ancestor around 510 million years ago. Plants had to make significant changes to their cellular, morphological, regulatory and physiological processes during their adaptation to the terrestrial environment from an aquatic environment. The first part of the project was to find out how these changes were reflected on the protein makeup of the early land plants. The gene model sequence data of two early land plants, Physcomitrella patens (moss) …

Is Metabolism Goal-Directed? Investigating The Validity Of Modeling Biological Systems With Cybernetic Control Via Omic Data, Frank T. Devilbiss

Open Access Dissertations

Cybernetic models are uniquely juxtaposed to other metabolic modeling frameworks in that they describe the time-dependent regulation of cellular reactions in terms of dynamic "metabolic goals." This approach contrasts starkly with purely mechanistic descriptions of metabolic regulation which seek to explain metabolic processes in high resolution — a clearly daunting undertaking. Over a span of three decades, cybernetic models have been used to predict metabolic phenomena ranging from resource consumption in mixed-substrate environments to intracellular reaction fluxes of intricate metabolic networks. While the cybernetic approach has been validated in its utility for the prediction of metabolic phenomena, its central feature, …

Influence Of The 3d Microenvironment On Glioblastoma Migration And Drug Response, Ruth Marisol Herrera Perez

Open Access Dissertations

Glioblastoma (GBM) is a highly invasive brain cancer characterized by poor prognosis. Despite significant efforts by the basic and clinical research community our understanding of GBM progression and recurrence has been incremental. Improvements in therapeutic response have been dismal, and GBM continues to be the deadliest tumor of the central nervous system, with patient average survival rate of 12 months. Synergistic relationships that the tumor cells establish with the brain microenvironment have been proven fundamental for successful tumor progression and maintenance. Yet, many in vitro GBM studies are performed in formats that fail to recapitulate the most essential component of …

Analysis Of Chd Remodelers During Development: A Tale In Two Organisms, Brett Bishop

Open Access Dissertations

The correct development of different organisms requires the precise timing of genes important for development transitions. Organisms have recruited ATP-dependent chromatin remodelers to ensure the correct timing of gene expression during developmental transitions. Here I show how different CHD ATP-dependent chromatin remodelers regulate developmental transitions of different organisms. I show that PICKLE not only promotes H3K27me3 during development to repress developmental genes but also is targeted to these genes. The association of PICKLE to these genes suggests that both repression and H3K27me3 levels is a direct action of PICKLE on these loci. Using zebrafish as a model system, I show …

Acrolein As A Novel Therapeutic Target For Spinal Cord Injury Induced Neuropathic Pain, Jonghyuck Park

Open Access Dissertations

Despite years of research, post-spinal cord injury (SCI) chronic neuropathic pain remains refractory to treatment and drastically impairs quality of life for SCI victims beyond paralysis. Although inflammation and free radicals contribute to neuropathic pain in SCI, the mechanism is not completely clear. We have recently demonstrated that acrolein, a product and catalyst of lipid peroxidation, induces a vicious cycle of oxidative stress, amplifying its effects and perpetuating oxidative stress and inflammation. In the current study, we have confirmed that acrolein is elevated significantly at least two weeks post-SCI which coincides with the emergence of hyperalgesia (mechanical, cold and thermal). …

Modulation Of Host Phosphatidylinositol Phosphates By Salmonella Effector Protein Sopb, Heather L. Piscatelli

Open Access Dissertations

Salmonella spp. are gram negative bacteria capable of infecting a number of eukaryotic hosts. In humans, Salmonella infection can range anywhere from acute gastroenteritis to typhoid fever which can oftentimes be fatal.Salmonella are facultative intracellular pathogens that have acquired the ability to enter non-phagocytic cells such as those lining the intestinal epithelium. Uptake into epithelial cells is mediated by the Salmonellapathogenicity island 1 (SPI1) encoded type III secretion system (T3SS), a needle-like complex composed of over 20 proteins that translocates effector proteins directly into the host cell cytosol. Salmonella possess a second type III secretion system encoded on …

Migratory Ecology Of Sea Turtles, Nathan Jack Robinson

Open Access Dissertations

Establishing the movement patterns of free-ranging animals is imperative to understanding their behavior and ecology, and is often necessary for designing effective conservation-strategies. This is especially true for migratory species, such as sea turtles, whose long-distance movements form a major component of their life history. In this thesis, I investigated which factors are driving the migratory behavior of the leatherback turtleDermochelys coriacea. Firstly, I examined whether the timing of the nesting season (nesting phenology) is influenced by oceanographic conditions along the pre-nesting migratory route or by variation in population structure. The discovery that nesting phenology appears more influenced by …

Role Of Group Ii Metabotropic Glutamate Receptor Subtype 2 (Mglur2) In Appetitive And Consummatory Aspects Of Ethanol Reinforcement, Kyle Allyson Windisch

Open Access Dissertations

Background: Group II metabotropic glutamate receptors (mGluR2/3) are predominately presynaptically located G_i/o coupled receptors that are highly expressed in the cortex, nucleus accumbens, amygdala, and hippocampus. Previous studies suggest that group II mGluRs are involved in regulating ethanol (EtOH) consumption and seeking following extinction (Backstrom and Hyytia, 2005; Kufahl, et al., 2011). The sipper tube model, which allows for procedural separation of seeking and consumption, was used to further clarify the role of mGluR2/3 in EtOH-seeking and consumption. The non-selective group II mGluR agonist LY379268 (LY37) and selective mGluR2 positive allosteric modulator (PAM) BINA were used to determine the …

Host Cell Death In Legionella Pneumophila Pathogenesis And Immunity, Wenhan Zhu

Open Access Dissertations

Legionella pneumophila is an intracellular pathogen that causes a severe, atypical pneumonia termed Legionnaires' disease. Upon entering the host cell, L. pneumophila resides in a membrane-bound vacuole, in which the bacterium evades lysosomal fusion and replicates. The establishment of the vacuole requires the Dot/Icm (Defect in organelle trafficking/ intracellular multiplication) transport system, which translocates a large number of substrates into host cells to re-orchestrate various cellular processes, such as intracellular trafficking, protein synthesis and host cell death pathways. Therefore, a key step in understanding the biology of Legionella is to dissect the mechanisms of action of the Dot/Icm substrates. By …

Nano-Engineered Polymers In Drug Delivery: Potential Approaches For Attenuation Of Secondary Injury After Spinal Cord Trauma, Wen Gao

Open Access Dissertations

Secondary injury elicits a complex series of pathophysiological events after the primary spinal cord trauma and even after its implantation treatment for neural functional recovery. These secondary injuries include an up-regulation of glial cells associated reactive oxygen species, nitrogen species, and reactive astrogliosis, and they can result in various levels of cellular and tissue damage. The inhibition of them has been proved to lead to functional recovery of the spinal cord. In this study, we concentrated on developing polymers and nano-techniques based drug delivery strategies to eliminate these secondary injuries. ^ To maintain and improve the performance of the implants …

Associated Behavioral, Genetic, And Gene Expression Variation With Alternative Life History Tactics In Salmonid Fishes, Ashley Chin-Baarstad

Open Access Dissertations

Individual differences in behavior can have potential fitness consequences and often reflect underlying genetic variation. My research focuses on three objectives related to individual level variation: 1) evaluating the innate behavioral variation within and between individuals, families, and progeny of different life-history types across time; 2) testing for differences in gene expression within the brain associated with this behavioral variation; and 3) using genetic polymorphisms to test for associations with ecotype, as well as population structure, in polymorphic populations. First, we evaluated the variation in a suite of ecologically relevant behaviors across time in juvenile progeny produced from crosses within …

Dietary Carbohydrates Influence The Structure And Function Of The Intestinal Alpha-Glucosidases, Mohammad Chegeni

Open Access Dissertations

As the primary products of starch digestion by pancreatic α-amylase, maltooligosaccharides (including maltose) are the main substrates for the α-glucosidases at the intestinal brush border. Here, maltose was shown to induce the formation of a higher molecular weight (HMW) sucrase-isomaltase (SI) species in Caco-2 cells that sorts more quickly to the enterocyte surface to act as a digestive enzyme. As this finding suggested a maltose sensing ability of small intestinal enterocytes, molecular mechanisms associated with the maturation and trafficking of HMW SI were further investigated. A pulse-chase experiment using [ ³⁵S]-methionine revealed a higher rate of early trafficking and …

Quantitative Modeling Of Spatiotemporal Systems: Simulation Of Biological Systems And Analysis Of Error Metric Effects On Model Fitting, James Hengenius

Open Access Dissertations

Understanding the biophysical processes underlying biological and biotechnological processes is a prerequisite for therapeutic treatments and technological innovation. With the exponential growth of computational processing speed, experimental findings in these fields have been complemented by dynamic simulations of developmental signaling and genetic interactions. Models provide means to evaluate "emergent" properties of systems sometimes inaccessible by reductionist approaches, making them test beds for biological inference and technological refinement.^ The complexity and interconnectedness of biological processes pose special challenges to modelers; biological models typically possess a large number of unknown parameters relative to their counterparts in other physical sciences. Estimating these parameter …

Towards A Paradigm Shift In The Modeling Of Soil Organic Carbon Decomposition For Earth System Models, Yujie He

Open Access Dissertations

Soils are the largest terrestrial carbon pools and contain approximately 2200 Pg of carbon. Thus, the dynamics of soil carbon plays an important role in the global carbon cycle and climate system. Earth System Models are used to project future interactions between terrestrial ecosystem carbon dynamics and climate. However, these models often predict a wide range of soil carbon responses and their formulations have lagged behind recent soil science advances, omitting key biogeochemical mechanisms. In contrast, recent mechanistically-based biogeochemical models that explicitly account for microbial biomass pools and enzyme kinetics that catalyze soil carbon decomposition produce notably different results and …

Intranuclear Strain Measured By Iterative Warping In Cells Under Mechanical And Osmotic Stress, Jonathan T Henderson

Open Access Dissertations

The nucleus is a membrane bound organelle and regulation center for gene expression in the cell. Mechanical forces transfer to the nucleus directly and indirectly through specific cellular cytoskeletal structures and pathways. There is increasing evidence that the transferred forces to the nucleus orchestrate gene expression activity. Methods to characterize nuclear mechanics typically study isolated cells or cells embedded in 3D gel matrices. Often report only aspect ratio and volume changes, measures that oversimplify the inherent complexity of internal strain patterns. This presents technical challenges to simultaneously observe small scale nuclear mechanics and gene expression levels inside the nuclei of …

Identification And Characterization Of Signal Components In Early Aba Signaling In Arabidopsis, Yueh-Ju Hou

Open Access Dissertations

The phytohormone abscisic acid (ABA) regulates growth, development, and response to abiotic stress in plants. The core ABA signaling has three key components: ABA receptors PYRABACTIN RESISTANCE 1 and PYR1-Like protein (PYR1/PYLs), type 2C protein phosphatases (PP2Cs) and SNF1- related protein Kinase 2s (SnRK2s). In the presence of ABA, PYR1/PYLs bind to and inhibit PP2Cs, leading to the release of SnRK2s from suppression, to activate downstream events such as ABA-responsive gene expression. To identify new components in the ABA signaling pathway, we performed a yeast two-hybrid screen for PYLs and SnRK2s interactors. We identified two proteins that interact with PYLs …

Identification And Characterization Of Factors Involved In Dna Demethylation And Anti-Silencing In Arabidopsis, Zhaobo Lang

Open Access Dissertations

DNA methylation is a key epigenetic mark for transcriptional gene silencing in many eukaryotes. DNA methylation status can be dynamically controlled by methylation and active demethylation processes. Compared to the well-known mechanisms of DNA methylation, the mechanisms of DNA demethylation and its regulation are poorly understood. In order to better understand the DNA demethylation pathway, we developed two genetic screening systems in Arabidopsis to identify new components involved in the DNA demethylation. In the first system, which is based on 35S promoter driven SUC2 (sucrose transporter 2) transgene, a mutant (91-1) was isolated and map-based cloning identified the …

Multidimensional Approach To Comparative Avian Visual Systems, Bret Alan Moore

Open Access Dissertations

Since the birth of visual ecology, comparative studies on how birds see their world have been limited to a small number of species and tended to focus on a single visual trait. This approach has constrained our ability to understand the diversity and evolution of the avian visual system. The goal of this dissertation was to characterize multiple visual dimensions on bird groups that are highly speciouse (e.g., Passeriformes), and test some hypotheses and predictions, using modern comparative tools, on the relationship between different visual traits and their association with visual information sampling behaviors. First, I developed a novel method …

Morphometric Characterization Of Individual Sympathetic Postganglionic Axons Innervating The Muscle Layers Of The Gastrointestinal Tract Of The Rat: A Complex Effector Model, Gary C. Walter

Open Access Dissertations

A full description of the terminal morphology of sympathetic postganglionic axons innervating the musculature of the gastrointestinal (GI) tract has not been available. Furthermore, common assumptions about the morphology and distribution of catecholaminergic terminal fields have been strongly shaped by the limitations of the techniques employed to distinguish the fibers and complicated by inconsistent findings generated with various methodologies. Thus, the present experiment used modern neural tracer techniques to provide high-resolution labeling of sympathetic fibers projecting to the smooth muscle wall of the GI tract. Fischer 344 rats (N = 50) received injections of dextran biotin into the left …

Population Differentiation, Dispersal Limitation, And Local Adaptation In The Gametophytic Fern Vittaria Appalachiana, Sally M. Stevens

Open Access Dissertations

Species' distributions are driven by a variety of abiotic and biotic factors. As these factors become altered by global climate change, species are believed to respond to these projected environmental changes in four different ways. One response is the shifting of the species' geographic range to higher latitudes and elevations, which will be unlikely for those species that have limited dispersal potential. Alternatively, organisms may tolerate the change, which will be unlikely for those organisms that are not phenotypically plastic. A third potential response is to adapt to the new environment via rapid evolution, an unlikely response for those organisms …

Integrative High-Throughput Study Of Arsenic Hyper-Accumulation In Pteris Vittata, Qiong Wu

Open Access Dissertations

Arsenic is a natural contaminant in the soil and ground water, which raises considerable concerns in food safety and human health worldwide. The fernPteris vittata (Chinese brake fern) is the first identified arsenic hyperaccumulator[1]. It and its close relatives have un-paralleled ability to tolerant arsenic and feature unique arsenic metabolisms. The focus of the research presented in this thesis is to elucidate the fundamentals of arsenic tolerance and hyper-accumulation in Pteris vittata through high throughput technology and bioinformatics tools. The transcriptome of the P. vittatagametophyte under arsenate stress was obtained using RNA-Seq technology and Trinity de novo assembly. …

Changes In Gene Expression And Viral Titer In Varroa Jacobsoni Mites After A Host Shift Asian To European Honey Bees, Gladys K. Andino Bautista

Open Access Dissertations

Honey bees (Apis mellifera L.) are the most important insects for the pollination of crops and wildflowers. However, they have experienced increasing colony die-offs during the past two decades. Multiple species of parasitic mites have been described that affect honey bees. The most important species in beekeeping belong to the genus Varroa ( Varroa jacobsoniand Varroa destructor). Varroa mite parasitism of honey bees is thought to be the most significant cause of colony mortality worldwide, and mite resistance to active ingredients of acaricides has become common. V. destructor causes direct impacts on bee production as well as …

Dj-1 And Atp13a2: Two Proteins Involved In Parkinson’S Disease, Josephat M Asiago

Open Access Dissertations

Parkinson's disease (PD) is the second most common neurodegenerative disorder after Alzheimer's disease, affecting approximately 0.3% of the total U.S. population, and its prevalence increases with age. Two neuropathological hallmarks of PD are the loss of dopaminergic neurons in the substantia nigra pars compacta, a region in the midbrain involved in initiating and sustaining movement, and the presence of cytosolic inclusions called Lewy bodies (LBs) in various brain regions. LBs are enriched with fibrillar forms of the presynaptic protein &agr;-synuclein (aSyn). Two autosomal recessive genes implicated in familial PD are PARK9, encoding the P-type ATPase ATP13A2, a lysosomal ATPase; and …

Characterization Of Caxck31, A Bacterial Calcium/Proton Antiporter, Marc Robert Ridilla

Open Access Dissertations

To better understand a class of transporters known as Calcium/Cation Antiporters (CaCAs), the bacterial calcium/proton antiporter CAXCK31 was purified and characterized. New methods were developed for its heterologous overexpression and purification. These methods help to define stress responses to toxic membrane overproduction in E. coli and may be broadly applicable to studies of membrane proteins. The results from a variety of biochemical and biophysical experiments demonstrated that CAXCK31 exists as a dimer in the membrane and can be purified in the dimeric state. The methods used include chemical cross-linking, FRET, and SEC-MALS. In addition, various transport properties of CAXCK31, including …