Life Sciences Commons^™

Utilization Of Ferrioxamine Microarrays For The Rapid Detection Of Pathogenic Bacteria, Nigam Bir Arora

Open Access Dissertations

Siderophores are low-molecular weight species utilized by bacteria for the sequestration of iron, an essential nutrient. Siderophores and their cognate receptors are considered to be virulence factors, due to their prominent role in pathogenicity. The work presented here focuses on ferrioxamine (FOx) as an “immutable” ligand for pathogen detection. A number of bacterial strains expressing high-affinity FOx receptors were identified by a proteomic BLAST search, and screened against microarrays patterned with FOx conjugates for detection using label-free optical imaging. Aspects such as inkjet printing and surface chemistry, iron-limiting conditions and bacterial selection protocols, and linker conjugate design were addressed and …

Investigating The Effects Of Ph On Alphaviral E3-E2 Glycoprotein Association, Organization, And Cellular Tropism, Jason Michael Sequra

Open Access Dissertations

In alphaviruses the role of E3 is required in protecting the fusion peptide region of E1 during intracellular transport. Throughout viral processing, the association of E2 and E3 is required for the successful trafficking and incorporation of E1 into the mature virion. This E3-E2 association has been observed to extend to mature virions in the solved structure for the envelope of Semliki Forest virus (SFV) and supported by the solved structure for the entire Venezuelan equine encephalitis virion (VEEV) with exclusive contacts being made between E3-E2. Immunization with monoclonal antibodies against VEEV E3 provided protection for mice challenged by lethal …

Complex Glycan Utilization Preferences Of Human Gut Bacteria, Yunus Emre Tuncil

Open Access Dissertations

Complex glycans, making up dietary fiber, have gained significance in recent years as they are the main energy source for the colonic microbiota which are physiologically important for the host health. Understanding glycan utilization strategies of the members of this community is essential to maximize beneficial health outcomes obtained from them. In order to investigate how the members of our gut microbiota utilize glycans and activate their respective polysaccharide utilization loci (PULs), we performed a series of time-course assays in which two model organisms, Bacteroides thetaiotamicron (Bt) and B. ovatus (Bo), were individually grown in a medium containing carbohydrates utilized …

Repurposing Non-Antimicrobial Drugs To Treat Multi-Drug Resistant Bacterial And Fungal Infections, Shankar Thangamani

Open Access Dissertations

Bacterial and fungal resistance to conventional antimicrobials is a burgeoning global health epidemic that necessitates urgent action. Even more alarming, the development of new antimicrobials to treat these multidrug-resistant pathogens has not kept pace with the rapid emergence of resistance to current antimicrobials. Antimicrobial drug development through the traditional de novo process is a risky venture given the significant financial and time investment required by researchers and limited success rate of translating these compounds to the clinical setting. This has led researchers to mine existing libraries of clinical molecules in order to repurpose old drugs for new applications (as antimicrobials). …

Mitigating Prevalence And Persistence Of L. Monocytogenes Using Novel Sanitation Strategies In Delis, Susan Rose Hammons

Open Access Dissertations

Ready-to-eat (RTE) deli meats sliced at retail are predicted to cause 83% of deli meat-associated listeriosis cases annually. While L. monocytogenes is commonly found in delis (55-90%), environmental prevalence varies by store (0-40%). Study 1 evaluated efficacy of 10 standardized SSOPs by testing 28 food and non-food contact surfaces for L. monocytogenes over 6 months in 30 retail delis. Intervention strategies employed in this study significantly decreased L. monocytogenesprevalence on FCS and NFCS in stores with moderate prevalence (0.5%, 0.8% reduction). However, we found that L. monocytogenes could persist despite implementation of SSOP control strategies. Study 2 assessed an …

Affinity Cryo-Electron Microscopy: Methods Development And Applications, Guimei Yu

Open Access Dissertations

Single particle cryo-electron microscopy (cryo-EM) is an emerging powerful tool for structural studies of macromolecular assemblies. Although less concentrated and smaller amounts of samples are required for single particle cryo-EM compared to X-ray crystallography, it remains challenging to study specimens that are low-abundance, low-yield, or short-lived. The recent development of affinity grid techniques holds great promise to tackle these challenging samples by combining the sample purification and freezing on TEM grids steps in cryo-EM grid preparation into a single step, revolutionize the grid preparation of cryo-EM, and extend single particle cryo-EM to a routine structural biology tool to characterize structures …

Magnesium Regulates Transcription Of The Mgta (Magnesium Transporter) Gene In Salmonella Enterica Serovar Typhimurium Via Prolyl-Bond Formation During Translation Of The Mgtl Leader Orf, Aaron R. Gall

Open Access Dissertations

In Enterobacteriaceae, the mgtA gene encodes a P-type ATPase that mediates Mg2+ uptake and is up-regulated by the PhoQP two-component system during invasion of host epithelial cells and macrophages. The regulation of mgtA has recently gained special interest given that it exists at several stages, including transcription, post-transcription and post-translation, in response to Mg2+availability. The mgtA mRNA has a 264-nucleotide 5’ leader that contains a 17 codon, proline-rich ORF, termed mgtL, whose translation has been proposed to affect the folding of the 5’ leader mRNA, which in turn regulates whether transcription is terminated before the mgtA structural gene at high …

Investigating Physiological Collaborations Between A Lower Termite And Its Symbionts, Brittany F. Peterson

Open Access Dissertations

This project was completed in an effort to better understand the contributions of symbiotic microbes to the biology of Reticulitermes flavipes, the eastern subterranean termite. Lower-termites, like R. flavipes, house symbionts from all three domains of life within their hindgut paunch. This intimate association is reflected in nearly every aspect of termite biology. Here, I investigate these physiological collaborations as they relate to digestion and immunity. My efforts focused on 1) quantifying the role of bacteria in wood digestion within the termite gut, 2) evaluating the role of symbionts in protection against pathogens, and 3) identifying gene products that bacterial …

Factors Affecting Transduction Efficiency Of Pseudotyped Viral Vectors Incorporating Alphaviral Glycoproteins, Aditi Kesari

Open Access Dissertations

The genome of an organism has the complete set of biochemical instructions required for sustenance of life. Mutations or abnormalities in this genome lead to genetic disorders. Currently available therapeutic options mostly focus on treating the symptoms, but not curing them. Gene therapy promises to be a curative form of medicine. In gene therapy cells carrying a defective gene are targeted and replaced with a healthy copy of that gene. The vehicles used for delivering this gene are known as vectors. Retroviruses are popularly used gene therapy/transfer vectors. However, retroviruses are limited in the range of cells they can enter …

Structural And Functional Studies Of The Papain-Like Protease 2 From Mouse Hepatitis Virus, Yafang Chen

Open Access Dissertations

Our goal is to establish a system to investigate how the deubiquitinating (DUB) and deISGylating activities of coronavirus (CoV) papain-like protease domains (PLPs) are involved in virus immune evasion. To this end, we chose PLP2 from mouse hepatitis virus (MHV) as our target of study because MHV has historically served as a model system for the study of CoVs, and it has undeniable advantage of ease in culturing in comparison to human coronaviruses.

It is reported here the expression and purification of a region of MHV nsp3 that contains the catalytic core of the PLP2 domain and its neighboring domains. …

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 …

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 …

Soil Quality And Nitrogen Cycling Dynamics Altered By Fertility Amendments In Intensively Managed Vegetable Production Systems, Matt A. Rudisill

Open Access Dissertations

Fresh market vegetable production in the Midwest U.S. is growing rapidly as a result of increased demand for locally-sourced food. High tunnels are becoming increasingly popular among Midwest vegetable growers as these structures offer many production benefits including season extension. However, vegetables require substantial fertility inputs, particularly nitrogen (N), in order to meet desired levels of productivity, and some fertility management strategies can negatively impact soil and environmental quality. This can be particularly problematic in high tunnels, where longer production periods and modification of the growing environment put these systems at greater risk for soil degradation. Identification of fertility practices …

Structural Studies On The Rubella Virus Capsid Protein And Its Organization In The Virion, Vidya Mangala Prasad

Open Access Dissertations

Rubella virus is a leading cause of birth defects due to infectious agents. When contracted during pregnancy, rubella infection leads to severe damage in fetuses. Despite its medical importance, very little is known about the structure of the pleomorphic rubella virus as compared to its alphavirus relatives. The rubella capsid protein is a critical structural component of virions as well as a key factor in virus-host interactions. Three crystal structures of the structural domain of the rubella capsid protein have been described here. The polypeptide fold of the capsid protomer has not been observed previously. The capsid protein structure, along …

Hybrid Opto-Electrokinetic Technique For Micro/Nanomanipulation: Towards Application Of A Novel Non-Invasive Manipulation Technique In Microbiological Assay, Jae-Sung Kwon

Open Access Dissertations

This dissertation explores various physical mechanisms of the Rapid Electrokinetic Patterning (REP) technique suggested for rapid and precise on-chip manipulation of colloids and fluids, and bio-compatibility of the technique for biological applications. REP is a hybrid opto-electrokinetic technique that is driven by the simultaneous application of an AC electric field and a heating source. It can not only effectively transport and manipulate a fluid but also concentrate and pattern particles suspended in the fluid through the combined effect of an electrohydrodynamic flow, electrostatic colloidal interactions and an electrothermal microfluidic flow. These capabilities make REP a promising tool which can provide …

Evidence Of An Infectious Asthma Phenotype: Chlamydia Driven Allergy And Airway Hyperresponsiveness In Pediatric Asthma, Katir Kirit Patel

Open Access Dissertations

Asthma is the most common chronic respiratory disease affecting young children and adults all over the world. An estimated 34.1 million Americans have reported asthma in their lifetime and the disease costs ~US $56 billion dollars to treat each year. Current treatment is based on a paradigm of asthma as a non-infectious atopic condition whose root cause is inflammation. Chronically administered anti-inflammatory medications, primarily inhaled corticosteroids (ICS), ameliorate asthma symptoms in many patients. However, up to 50% of asthmatics, characterized by neutrophil infiltration, IL-17 secretion and increased risk of fatality are refractory to ICS treatment. Chlamydia pneumoniae, a ubiquitous, obligate …

Reca Dynamics & The Sos Response In Escherichia Coli: Cellular Limitation Of Inducing Filaments, Shawn Christopher Massoni

Open Access Dissertations

During the course of normal DNA replication, replication forks are constantly encountering "housekeeping" types of routine damage to the DNA template that may cause the forks to stall or collapse. One product of this fork collapse is the induction of the SOS response, a coordinated global response to help pause the growth and replication of a cell while DNA damage is addressed and repaired. In E. coli, this response is activated by the formation of ssDNA, to which the RecA protein binds and forms a nucleoprotein filament, which acts as the activator for autocleavage of the LexA transcriptional repressor, which …

Investigation Of A Sulfur-Utilizing Perchlorate-Reducing Bacterial Consortium, Teresa Anne Conneely

Open Access Dissertations

We present research investigating how, with in depth knowledge of the community, microbial communities may be harnessed for bioremediation of hazardous water contaminants. We focused on the bacterial reduction of perchlorate, a common water contaminant. For this we studied the structure and capabilities of a novel sulfur-utilizing, perchlorate-reducing bacterial (SUPeRB) consortium. Initially, we characterized the minimal consortium that retained functional capabilities, using 16S rRNA and functional gene analysis. A diverse functional consortium dominated by Beta-Proteobacteria of the family Rhodocyclaceae and sulfur-oxidizing Epsilon-Proteobacteria was found. We also examined the optimal growth conditions under which perchlorate degradation occurred and uncovered the upper …

Mitochondrial Dna Polymerase Ib: Functional Characterization Of A Putative Drug Target For African Sleeping Sickness, David F. Bruhn

Open Access Dissertations

Trypanosoma brucei and related parasites are causative agents of severe diseases that affect global health and economy. T. brucei is responsible for sleeping sickness in humans (African trypanosomiasis) and a wasting disease in livestock. More than 100 years after T. brucei was identified as the etiological agent for sleeping sickness, available treatments remain inadequate, complicated by toxicity, lengthy and expensive administration regiments, and drug-resistance. There is clear need for the development of a new antitrypanosomal drugs. Due to the unique evolutionary position of these early diverging eukaryotes, trypanosomes posses a number of biological properties unparalleled in other organisms, including humans, …

Growth Kinetics And Constraints Related To Metabolic Diversity And Abundances Of Hyperthermophiles In Deep-Sea Hydrothermal Vents, Helene Chavanne Ver Eecke

Open Access Dissertations

This dissertation research aims to show that there are deterministic microbial distribution patterns based on quantifiable environmental thresholds by determining and rationalizing the relative abundances of hyperthermophilic methanogens, autotrophic iron(III) oxide reducers, and heterotrophic sulfur reducers within deep-sea hydrothermal vents. Organisms of these metabolisms are predicted to be relatively more abundant in different regions depending on environmental conditions such as reduction potential, organic carbon, and hydrogen availability. The relative abundances of these metabolic groups within samples from the Endeavour Segment and Axial Volcano in the northeastern Pacific Ocean were determined. Iron(III) oxide reducers were detected in nearly all samples while …

Role Of Sulfate-Reducing Bacteria In The Attenuation Of Acid Mine Drainage Through Sulfate And Iron Reduction, Caryl Ann Becerra

Open Access Dissertations

Acid mine drainage (AMD) is an acidic, iron-rich leachate that causes the dissolution of metals. It constitutes a worldwide problem of environmental contamination detrimental to aquatic life and water quality. AMD, however, is naturally attenuated at Davis Mine in Rowe, Massachusetts. We hypothesize that sulfate-reducing bacteria (SRB) are attenuating AMD. To elucidate the mechanisms by which SRB attenuate AMD, three research projects were conducted using a suite of molecular and geochemical techniques. First we established biological influence on the attenuation of AMD by comparing the microbial community and geochemical trends of microcosms of two contrasting areas within the site: AMD …

Investigation Of Chemotaxis Genes And Their Functions In Geobacter Species, Hoa T. Tran

Open Access Dissertations

Geobacter species are δ-Proteobacteria and are often predominant in the Fe(III) reduction zone of sedimentary environments. Their abilities to remediate contaminated environments and to produce electricity have inspired extensive studies. Cell motility, biofilm formation, and type IV pili, which have been shown to be regulated by chemotaxis genes in other bacteria, all appear important for the growth of Geobacter species in changing environments and for electricity production. The genomes of Geobacter species show the presence of a significant number of chemotaxis gene homologs, suggesting important roles for them in the physiology of Geobacter species, although gene functions are not yet …