Life Sciences Commons^™

Characterizing The Human Vaginal Microbiome Using High-Throughput Sequencing, Jean Megan E. Macklaim

Electronic Thesis and Dissertation Repository

The human vaginal microbiome undoubtedly has a significant role in reproductive health and for protection from infectious organisms. Recent efforts to characterize the bacterial species of the vagina using molecular techniques have uncovered an unexpected diversity. Using high-throughput sequencing I sought to describe the structure and function of the vaginal microbiome under different physiological states including healthy, bacterial vaginosis (BV), post-menopausal vaginal atrophy, and acute vulvovaginal candidiasis (VVC).

Partial 16S rRNA gene sequencing revealed that healthy, asymptomatic women most often have vaginal biotas dominated by Lactobacillus iners or L. crispatus. In contrast, BV is a heterogeneous, highly diversified condition …

Effects Of Biodiesel Concentration On Microbial Deterioration Of Polyethylene In A Simulated Fuel Storage Tank, Juan Manuel Restrepo-Florez

Electronic Thesis and Dissertation Repository

In this study a simulated fuel storage tank was used to investigate the effect of biodiesel concentration on biodegradation of polyethylene. This research is relevant in the field of fuel storage. The simulated storage system consisted of a number of identical conical flasks. Each flask was comprised of two layers, an upper one consisting of a fuel blend of diesel with biodiesel in concentrations ranging from 0 to 100% of biodiesel and the bottom layer containing an aqueous mineral media inoculated with a community obtained from a real fuel storage facility. Polyethylene slabs cut to a specific size were immersed …

Iron-Regulated Cyanobacterial Predominance And Siderophore Production In Oligotrophic Freshwater Lakes, Ryan J. Sorichetti

Electronic Thesis and Dissertation Repository

The frequency and intensity of cyanobacterial blooms (cyanoblooms) is increasing globally. Contrary to existing phosphorus (P) and nitrogen (N) paradigms describing cyanobloom proliferation in eutrophic (nutrient-rich) freshwater lakes, many of the recent cyanobloom reports pertain to oligotrophic (nutrient-poor) freshwater lakes with no prior history of cyanobloom occurrence. There exists a critical research need to re-visit existing conceptual models, identify regulating factors currently unaccounted for and improve our ability to effectively detect and measure cyanobacterial toxins (cyanotoxins) in lakes. Iron (Fe) is required in nearly all pathways of cyanobacterial macronutrient use, though its direct role in regulating cyanobacterial biomass is not …

Heterologous Production And Characterization Of Cell Wall Degrading Enzymes Using Plants As A Bioreactor, Eridan Orlando Rodrigues Pereira

Electronic Thesis and Dissertation Repository

Plants are wonderful living organisms. They are able to store solar energy into carbohydrates by fixing CO₂ through photosynthesis which can be subsequently harvested and used for fuel production. However, one of the major limitations for transforming these carbohydrates into liquid fuels is the recalcitrance of the plant cell wall. Although microorganisms have evolved a series of cell wall degrading enzymes to harvest efficiently this energy and are considered the main source of these biocatalysts, harnessing these microorganisms for the production of enzymes is a costly process and a major factor limiting the commercialization of lignocellulosic biomass-to-ethanol processes. The …

Pegylation As A Novel Tool To Investigate The Topology Of Escherichia Coli Weca, A Membrane Enzyme Involved In Lipopolysaccharide O Antigen Initiation, Stéphanie L. Lamothe

Electronic Thesis and Dissertation Repository

O-antigen, the most surface exposed moiety of bacterial lipopolysaccharide (LPS), plays several roles in pathogenicity. The biosynthesis of O-antigen starts by the formation of a phosphoanhydride bond linking a sugar phosphate with a membrane isoprenoid lipid phosphate. Two distinct families of integral membrane proteins catalyze this reaction. The protein WecA is the prototypic member of one of these families, termed the polyisoprenyl-phosphate N-acetylaminosugar-1-phosphate transferase (PNPT) family. Because the donor nucleotide sugar is only available in the cytosol, cytosolic exposed regions of WecA are expected to be critical for the catalytic activity of the enzyme. Therefore, elucidating an accurate topological …

Characterization Of Staphylococcus Aureus Lipase, Vithooshan Vijayakumaran

Electronic Thesis and Dissertation Repository

USA300, a strain of community-associated methicillin resistant Staphylococcus aureus (CA-MRSA), has become prevalent in the community. Colonization of human skin requires mechanisms that allow this bacterium to overcome the innate immune defenses on the skin, including secretion of antimicrobial lipids. Antimicrobial lipids inhibit S. aureus growth and induce the staphylococcal proteolytic cascade, producing aureolysin (Aur) which processes the lipase glycerol ester hydrolase (Geh). Nearly all S. aureus strains secrete Geh, yet little information exists concerning its function. Using purified Aur and Geh we confirm that aureolysin processes proGeh to Geh. We then confirmed that geh was required for lipase activity …

Role Of Branched-Chain Amino Acid Transporters In Staphylococcus Aureus Virulence, Sameha Omer

Electronic Thesis and Dissertation Repository

Branched-chain amino acids (BCAAs) act as effector molecules that signal a global transcriptional regulator, CodY, to regulate virulence factors in nutrient depleted environments. Staphylococcus aureus contains three putative BCAA transporters (BrnQ1, BrnQ2, BrnQ3) whose role in BCAA uptake is unknown. We hypothesize that BrnQ transporters are involved in BCAA uptake and contribute to virulence in S. aureus by modulating CodY activity. Results from radioactive uptake assays indicate that BrnQ1 is the predominant BrnQ transporter of isoleucine, valine and leucine. Meanwhile, BrnQ2 is more specific for isoleucine. Furthermore, only the lack of BrnQ1 hinders growth of S. aureus in chemically-defined media …

The Interaction Of The Human Adenovirus E1a Protein With The Human Dref Transcription Factor, Kris M. James

Electronic Thesis and Dissertation Repository

The human adenovirus (HAdV) E1A protein is the first protein produced post-HAdV infection, and serves two main functions. The first is to modulate host and viral transcription. The second is to induce host cell cycle progression to S phase, to promote an optimal environment for viral replication. E1A performs its functions by binding and manipulating over 50 cellular factors. Interestingly, I found that E1A is capable of interacting with the poorly characterized human DNA replication-related element-binding factor (hDREF). hDREF is a transcription factor associated with the expression of several genes related to the cell cycle. I hypothesized that the interaction …

Human Adenovirus E1a Binds And Retasks Cellular Hbre1, Blocking Interferon Signalling And Activating Virus Early Gene Transcription, Gregory J. Fonseca

Electronic Thesis and Dissertation Repository

Upon infection, human adenovirus (HAdV) must block interferon signaling and activate the expression of its early genes to reprogram the cellular environment to support virus replication. During the initial phase of infection, these processes are orchestrated by the first HAdV gene expressed during infection, early region 1A (E1A). E1A binds and appropriates components of the cellular transcriptional machinery to modulate cellular gene transcription and activate viral early genes transcription. We have identified hBre1/RNF20 as a novel target of E1A. hBre1 is an E3 ubiquitin ligase which acts with the Ube2b E2 conjugase and accessory factors RNF40 and WAC1 to monoubiquitinate …

Systematic Analysis Of Residues In Conserved Region 3 Of The Human Papillomavirus 16 E7 Oncoprotein, Biljana Todorovic

Electronic Thesis and Dissertation Repository

Although remarkable biological diversity is exhibited by viruses, as obligate intracellular parasites, they rely on host cell functions. As such, viruses typically must overcome a set of host barriers that prevent infection. For human papillomaviruses (HPV) one of these barriers is the state of terminal differentiation of the host cell. For that purpose HPVs encode two major oncoproteins, E6 and E7, which combine their efforts to effectively uncouple cellular differentiation from the cell cycle arrest. The E7 proteins have no intrinsic enzymatic activity or DNA binding ability, but they bind and manipulate numerous host proteins. E7 is a modular oncoprotein …

An Investigation Of Plant-Microbe Interactions Under Cadmium Stress In Agar-Based Medium, Hydroponics, And Soil Studies, Melanie P. Columbus

Electronic Thesis and Dissertation Repository

This thesis investigated plant-microbe-metal interactions at two scales: a single plant-microbe system and an agricultural rhizobacterial community. The first objective was to investigate the effectiveness of a plant growth-promoting rhizobacterium (PGPR) on mediating cadmium stress in a plant model system. Arabidopsis thaliana Col-0 was inoculated with Pseudomonas putida UW4, which in its wild type form has been reported to reduce plant stress by simultaneously metabolizing the ethylene precursor 1-aminocyclopropane-1-carboxylate (ACC) with the enzyme ACC deaminase and stimulating plant growth through the production of indole-3-acetic acid (IAA). A mutant strain that lacks ACC deaminase and a no bacteria treatment were used …