Life Sciences Commons^™

Novel Thermophilic Bacteria Isolated From Marine Hydrothermal Vents, Christine Demko Sislak

Dissertations and Theses

As part of a large study aimed at searching for patterns of diversity in the genus Persephonella along the north to south geochemical gradient of the ELSC, ten novel strains of Alphaproteobacteria were isolated unexpectedly. Using defined media under microaerophilic conditions to enrich for Persephonella from chimney samples collected at the seven vent fields on the ELSC and the dilution to extinction by serial dilution method to purify cultures, a total of ten strains belonging to the Alphaproteobacteria were isolated. Two of these isolates, designate MN-5 and TC-2 were chosen for further characterization and are proposed as two new species …

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 …

Identification Of The Binding Partners For Hspb2 And Cryab Reveals Myofibril And Mitochondrial Protein Interactions And Non-Redundant Roles For Small Heat Shock Proteins, Kelsey Murphey Langston

Theses and Dissertations

Small Heat Shock Proteins (sHSP) are molecular chaperones that play protective roles in cell survival and have been shown to possess chaperone activity. As such, mutations in this family of proteins result in a wide variety of diseases from cancers to cardiomyopathies. The sHSPs Beta-2 (HspB2) and alpha-beta crystalline (CryAB) are two of the ten human sHSPs and are both expressed in cardiac and skeletal muscle cells. A heart that cannot properly recover or defend against stressors such as extreme heat or cold, oxidative/reductive stress, and heavy metal-induced stress will constantly struggle to maintain efficient function. Accordingly, CryAB is required …

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 …

The Glycine And Proline Reductase Systems: An Evolutionary Perspective And Presence In Enterobacteriaceae, Joshua Witt

HIM 1990-2015

The Glycine and Proline Reduction systems are two of the best characterized selenoenzymes in bacteria and have been found to occur in a wide variety of clostridia [1-5]. These enzymes are utilized to reduce glycine or D-proline to obtain energy via substrate level phosporylation or membrane gradients, respectively [6, 7]. This includes the pathogens C. difficile and C. botulinum [5, 8]. Strains of C. difficile are activate toxigenic pathways whenever either of these pathways is active within the cell [5, 8]. Though evolutionary studies have been conducted on ammonia producing bacteria [9] none has been done to directly characterize these …

Wood Decomposition In A Warmer World, Emily Elizabeth Austin

Doctoral Dissertations

Climatic warming is altering species distributions and ecosystem functions across the globe. Wood is an important carbon pool and the fungal communities in wood are relatively simple compared to those in soil. These factors make decomposing wood an ideal system for exploring the influence of decomposer community on the response of decomposition to warming. My research has focused on the effects of warming wood decomposition rates and wood decomposing communities. Using field and lab- based manipulative experiments and field observations I explore the influence of tree species, wood decomposition stage, geography and warming on fungal community structure and activity. In …

The Incidence Of Antibiotic Resistance In Mesophilic Aeromonas Isolated From The Buffalo River And From A Non-Urban Site Upstream, Amy L. Chapman

Biology Theses

Antibiotics are commonly used in agriculture and industry and their discharge is commonly seen in rivers, like the Buffalo River. This antibiotic discharge may cause a selective environment which favors the growth of antibiotic resistant Aeromonas. To study the effect of urban pollution on the antibiotic resistance in Aeromonas, 229 Aeromonas isolates were collected from fish tissues as well as sediment and water samples collected from the Buffalo River and a non-urban site (Cazenovia Creek). Seven different Aeromonas taxa were identified using biochemical tests. There were 124 (54%) isolates that were classified as atypical, which was the most commonly …

Identification And Characterization Of Genetic Factors Involved In Candida-Bacterial Interactions, Sean J. Fox

Electronic Theses and Dissertations

Throughout existence, fungi and bacteria have long shared ecological niches and thus engage in numerous interactions to mutually enhance survival or antagonistically gain competitive advantages. Of importance to human health are those interactions that involve bacteria with the opportunistic fungi, Candida albicans. An important virulence factor of C. albicans is the ability to control morphology, which allows the transition between yeast, pseudohyphal, and hyphal phenotypes. Morphological control in C. albicans is governed by quorum sensing and the secreted autoregulatory molecule farnesol. Quorum sensing allows individual cells to sense the environment and respond as a group. Bacteria also use quorum …

Characterization Of The Role(S) Of Env Protein Of Human Endogenous Retrovirus-K In Multiple Human Cancers, Ming Li

Dissertations & Theses (Open Access)

Human endogenous retroviruses (HERVs) are remnants from ancient retroviral infections, and most of them are inactive in normal tissues. One family of HERV, HERV-K, is found to be associated with multiple human cancers including breast cancer, pancreatic cancer and melanoma, but the causal relationship between HERV-K and cancer is still unclear. Increased expression of HERV-K in melanoma cells correlates with malignant transformation, while a serological response to HERV-K in breast cancer or melanoma patients correlates with survival probability. However, the mechanism behind these observations remains obscure. Our laboratory reported that anti-HERV-K envelope (Env) protein antibodies show antitumor potential in targeting …

Validation Of Thermal Destruction Of Pathogenic Bacteria In Rendered Animal Products, Mary Hayes

All Dissertations

This study tested enumeration techniques for high fat food matrices and determined thermal death times in rendering animal products. Standard Class O phosphate/magnesium chloride dilution buffer series (Dilution Series A) and a modified (pre-warmed to 32oC) lecithin phosphate dilution buffer series (Dilution Series B) were used to enumerate a Salmonella cocktail from both poultry and beef rendering materials. Results of this study indicate use of a modified lecithin buffer did not improve Salmonella enumeration accuracy from rendering materials. Instead, the results suggested use of xylose lysine deoxycholate agar (XLD) with either buffer system accurately enumerated Salmonella from rendering materials. The …

Studies Of Molecular Mechanisms Of Cranberry Mediated Healthspan Promotion In Caenorhabditis Elegans, Sujay Guha

All Dissertations

Extensive studies have demonstrated the potent ability of many nutraceuticals to alleviate the symptoms of aging and stress. We found that cranberry extract (CBE) supplementation at an optimum concentration could prolong Caenorhabditis elegans (C. elegans) lifespan in a manner of daf-16 and osr-1 dependence. Considering that aging is a progressive degeneration of physiological functions, it is intriguing to know whether CBE consumption may delay the age-related degenerative changes in animals. Thus, we examined the effect of CBE consumption on healthspan in C. elegans populations with different ages and age-related disorders. Our results suggest that long-term CBE supplementation leads to a …

A Synthetic Biology Approach To Engineering New Anticancer Agents, Shane Robert Wesener

Theses and Dissertations

Histone deacetylase (HDAC) inhibitors are becoming increasingly valuable therapeutic agents in treatment of several types of malignancies. FK228 is a depsipeptde anticancer compound produced by Chromobacterium violaceum no. 968 through a nonribosomal peptide synthetase (NRPS)-polyketide synthase (PKS) hybrid assembly line. In the present study, reconstitution of the biosynthetic pathway responsible for the production of FK228 revealed cross-talk between modular PKS and fatty acid synthase. This pathway contains two PKS modules on the DepBC enzymes that lack a functional acyltransferase (AT) domain, and no apparent AT-encoding gene exists within the gene cluster or its vicinity. We reported through heterologous expression of …

C-Jun N-Terminal Kinases Regulate Adenovirus-Mediated Autophagy And Antigen Presentation, Sarah R. Klein

Dissertations & Theses (Open Access)

Targeted immunotherapy with recombinant, oncolytic adenoviruses is under investigation for the treatment of cancer. Evidence indicates adenoviruses induce autophagy that is required for oncolysis, but the molecular regulation of autophagy in infected cells remains under investigation. Our data suggested the canonical pathway regulating starvation-induced autophagy was not implemented in adenovirus-induced autophagy; however, adenovirus infection triggered phosphorylation of c-Jun N-terminal kinases (JNK) that was essential for autophagy. Adenoviral replication within the host cell elicited JNK pathway activation leading to B cell lymphoma-2 (Bcl-2) phosphorylation. JNK-dependent Bcl-2 phosphorylation stimulated the dissociation of Bcl-2/beclin 1 heterodimers, enabling beclin 1 to initiate autophagy. Moreover, …

Bacterially-Mediated Formation Of Rock Coatings In Kärkevagge, Swedish Lapland: A Mineralogical And Micro-Environmental Analog For Mars, Cassandra L. Marnocha

Graduate Theses and Dissertations

The search for past or present life on Mars is, for now, limited to surface environments. An often neglected surface environment that could have served as an abode for life and could presently preserve evidence of that life is that of rock coatings. Rock coatings are mineral accretions on rock surfaces. On Earth, they are widespread and occur with considerable chemical diversity. There is growing evidence for a biotic role in their formation on Earth, particularly with respect to rock varnish. As a result, rock varnish has become a target of astrobiological interest on Mars, where varnish-like coatings have been …

Proximity-Labeling Of Near Neighbors Of Lamin A And Lamin A-Δ50 (Progerin)., Mohammad Sabri

Undergraduate Honors Theses

In an attempt to isolate and identify proteins that differentially interact with or locate near lamin A and progerin, we used a previously described method named BioID (proximity-dependent biotin identification). This method is based on fusion of a promiscuous E. coli biotin-protein ligase (BL) to a targeting protein (in this study, lamin A-GFP and progerin-GFP). The biotin ligase biotinylates amino residues in proteins that are near-neighbors of the fusion protein. To create the fusion proteins, BL was sub-cloned from a pcDNA3.1 MCS-BirA(R118G)-HA plasmid donated by Kyle Roux from University of South Dakota. The BL fragment was ligated into a pNEBR-X1-lamin …

Environmental Constraints On Cyanomyophage Abundance In The Subtropical Pacific Ocean, Tiana Maria Pimentel

Masters Theses

Viruses are abundant in the world’s oceans and are thought to be important participants in marine biogeochemical cycling. Of these viruses, cyanophages are considered especially important because they infect and lyse cyanobacteria, which are some of the main primary producers in marine environments. Cyanophages are thought to influence the abundance and diversity of cyanobacterial populations and impart significant mortality, thereby affecting primary productivity and microbial community structure. Despite their ecological relevance, little is known about how environmental factors shape cyanophage abundance and diversity over large temporal and spatial scales. To address this gap in knowledge, seawater samples were collected during …

Investigating Potential Bioactive Compounds From Rhodococcus And Their Effects On Mcf7 Breast Cancer Cells, Megan N. Crabtree

Electronic Theses and Dissertations

Many drugs used in the treatment of various cancers are derived from or influenced by compounds from nature. The soil bacterium Rhodococcus is of interest because of its identified secondary metabolic pathways and the production of novel natural antibiotics from several strains. In this study, a solid agar extraction method was used to collect compounds from strains of Rhodococcus. These bacterial compound extracts were then tested using a MTT assay in order to evaluate their effectiveness in augmenting MCF7 breast cancer cell death. The results of two way ANOVA analyses revealed 18 compound extracts from 15 strains of Rhodococcus that …

The Performance Of Bacterial Phytosensing Transgenic Tobacco Under Field Conditions, Michael Harrison Fethe

Masters Theses

Currently the platforms for wide-area detection of environmental contamination are limited. Therefore, there is interest in developing new platforms, especially for use in crop plants to detect and report the presence of biotic and abiotic stress agents. A biosensor uses a biological organism or substrate to detect the presence of an elicitor (i.e., heavy metal, TNT, or bacteria). The foundational groundwork to create biosensors in transgenic plants exists. The creation of bacterial phytosensing transgenic tobacco containing an orange fluorescent protein (OFP) reporter driven by synthetic pathogen-inducible promoters provides a fluorescent signal when infected with phytopathogens for earlier detection in the …

The Influence Of Hydrogeomorphology, Soil Redox Conditions, And Salinity On The Spatial Zoning Of Saltgrass, Salt Rush, And Cattails In Scotts Creek Marsh, Swanton Pacific Ranch, Ca, Mark D. Gormley

Master's Theses

Scotts Creek Marsh (SCM) is a small coastal wetland ecosystem in Davenport, CA. The vegetation of SCM is dominated by three halophytic zones comprised of saltgrass, salt rush, cattails. The objectives of the study were (i) to investigate the variables that influence the zoning of the three dominant halophyte communities in SCM and (ii) to the test the effectiveness of Indicator of Reduction in Soil (IRIS) tubes to indicate the reduction of S. The study examined the following parameters from April 6 to July 21, 2013: (i) the HGM of Scotts Creek Marsh, (ii) soil oxidation and reduction (redox) conditions, …

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 …

The Effect Of Media Selection On The Membrane Characteristics And Fatty Acid Composition Of Staphylococcus Aureus, Seth Russell Johnson

Theses and Dissertations

The cellular membrane is a vital structure to a microorganism and impacts important aspects of environmental regulation and growth. The physical structure of cellular membranes plays a key role in bacterial growth at low temperatures, membrane permeability and susceptibility to membrane active molecules as well as broader aspects of bacterial physiology and pathogenesis. Mueller Hinton medium is the required medium for antimicrobial susceptibility testing in the US, but there are many other types of media used in staphylococcal research. Different media vary in their nutrient compositions, and the effect of media composition on bacterial physiology is incompletely understood. The aim …

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 …

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 …

A Quadruplex Real-Time Pcr Assay For The Rapid Detection And Differentiation Of The Burkholderia Pseudomallei Complex: B. Mallei, B. Pseudomallei, And B. Thailandensis, Chinn-Woan Lowe

Theses and Dissertations

Methods for the rapid detection and differentiation of the Burkholderia pseudomallei complex comprising B. pseudomallei, B. mallei, and B. thailandensis, have been the topic of recent research due to the high degree of phenotypic and genotypic similarities of these species. B. pseudomallei and B. mallei are the causative agents of melioidosis and glanders, respectively. B. pseudomallei and B. mallei are recognized by the CDC as tier 1 select agents. Although B. thailandensis is generally avirulent in mammals, this species displays very similar phenotypic characteristics to that of B. pseudomallei. Optimal identification of these species remains problematic, …

Regulation Of Hyaluronate Lyase (Hyla) Expression By Regr A Transcriptional Repressor In Streptococcus Pyogenes, Alexis A. Kordis

Biological Sciences Theses & Dissertations

Streptococcus pyogenes, group A streptococci (GAS), causes diseases ranging from asymptomatic to life threatening. Some strains of S. pyogenes produce extracellular hyaluronate lyase (HylA), a potential virulence factor. HylA is an enzyme that degrades hyaluronic acid, which is found in the extracellular matrix of human tissues. The breakdown of the host tissue contributes to the spread of infection. The hylA gene is not constitutively expressed in vitro, which implies regulation.

A proposed regulator of hyaluronate lyase expression is RegR, a LacI/GalR like protein. A vector containing disrupted regR was electro-transformed into S. pyogenes M-type 22 strain 10403. Growth characteristics …

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 …