Life Sciences Commons^™

Principles Of Aaa+ Proteases, Samar Mahmoud

Doctoral Dissertations

ATPases associated with diverse cellular activities (AAA+) proteases in bacteria help maintain protein homeostasis by degrading misfolded and regulatory proteins. While a handful of protein targets for these proteases have been identified in Caulobacter crescentus and other organisms, more research is needed to elucidate mechanisms that govern substrate specificity. In the second chapter of this thesis, I will elaborate on how AAA+ substrate specificity is less rigid than previous work has suggested and how limiting ATP or mutations can alter substrate preferences of the ClpXP protease. In the third chapter, I will highlight our efforts to use a quantitative proteomics …

Antibiotic Sensitivity Testing Of Foodborne Bacteria Using Surface-Enhanced Raman Spectroscopy, Joshua Gukowsky

Doctoral Dissertations

The spread of antibiotic resistant bacteria around the world has become a major public health issue, and it is essential that effective detection methods exist for identifying these organisms and preventing them from spreading throughout our food systems and into the environment. The goal of this research is to develop a novel analytical procedure that is capable of easily identifying antibiotic resistance in bacterial samples, and also provides more information about the biochemical characteristics of the bacteria and their responses to antibiotic exposure. Surface-enhanced Raman Spectroscopy (SERS), an analytical technique that uses light scattering to produce a spectrum based on …

Engineered Nanoparticles For Site-Specific Bioorthogonal Catalysis: Imaging And Therapy, Riddha Das

Doctoral Dissertations

Bioorthogonal catalysis offers a strategy for chemical transformations complementary to bioprocesses and has proven to be a powerful tool in biochemistry and medical sciences. Transition metal catalysts (TMCs) have emerged as a powerful tool to execute selective chemical transformations, however, lack of biocompatibility and stability limits their use in biological applications. Incorporation of TMCs into nanoparticle monolayers provides a versatile strategy for the generation of bioorthogonal nanocatalysts known as “nanozymes”. We have fabricated a family of nanozymes using gold nanoparticles (AuNPs) as scaffolds featuring diverse chemical functional groups for controlled localization of nanozymes in biological environments, providing unique strategies for …

Surface-Enhanced Raman Spectroscopic Imaging Of Bacteria Within Fresh Produce In Situ, Michael Hickey

Doctoral Dissertations

The growth curves for E. coli O157:H7 (#043888) are reported. We make the case that the onset of stationary growth is the optimal point at which a bacteria culture is considered suitable for quantitative Raman analyses. The optimal conditions for 3-mercaptophenylboronic acid coating of bacteria cells is also reported. Fundamental drawbacks of the status-quo approach have been elucidated and overcome, based on measurable improvements to the experimental methodology. This approach is shown to be suitable for the evaluation of bacterial rinse-washing efficacy by means of Raman light-scattering. The data were compared to label-free applications and the measurable differences between each …

Development Of Methodology For Rapid Bacterial Detection In Complex Matrices Using Sers, Madeline Tucker

Masters Theses

Fresh foods, including meats and produce are the fastest growing market in the supermarket and the class of foods most likely to cause a bacterial foodborne illness. As the rate of consumption of perishable products increases, rapid detection of pathogens within the food supply becomes a critical issue. Current methods used for the detection of bacteria that cause food-borne illnesses are time consuming, expensive and often require selective enrichment. In this study we adapted a separation technique originally developed for PCR to extract bacteria from ground beef using β-cyclodextrin (β-CD) and milk protein coated activated carbon (MP-CAC) as filtration agents. …

Structure-Property Relationships Of Polymer Films And Hydrogels To Control Bacterial Adhesion, Kristopher W. Kolewe

Doctoral Dissertations

The emergence and spread of antibiotic resistance across microbial species necessitates the need for alternative approaches to mitigate the risk of infection without relying on commercial antibiotics. Biofilm-related infections are a class of notoriously difficult to treat healthcare-associated infections that frequently develop on the surface of implanted medical devices. As biofilm formation is a surface-associated phenomenon, understanding how the intrinsic properties of materials affect bacterial adhesion enables the development of structure-property relationships that can guide the future design of infection-resistant materials. Despite lacking visual, auditory, and olfactory perception, bacteria still manage to sense and attach to surfaces. Previously, it has …

Pore Forming Protein Assembly And The Use In Nanopore Sensing: A Study On E. Coli Proteins Clya And Ompg, Monifa Fahie

Doctoral Dissertations

Pore forming proteins are typically the proteins that form channels in membranes. They have several roles ranging from molecule transport to triggering the death of a cell. This work focuses on two E. coli pore forming proteins that have vastly differing roles in nature. Outer membrane protein G (OmpG) is an innocuous β-barrel porin while Cytolysin A (ClyA) is an α-helical pore forming toxin. For OmpG we probed its potential to be a nanopore sensor for protein detection and quantification. A small high affinity ligand, biotin, was covalently attached to loop 6 of OmpG and used to capture biotin-binding proteins. …

The Effect Of Management Practices On Bacterial, Fungal, And Nematode Communities On Cool Season Turfgrass, Elisha Allan-Perkins

Doctoral Dissertations

Golf courses comprise 50 million acres in the United States of highly managed turf susceptible to abiotic and biotic stressors. A growing area of interest is utilizing microbes to improve plant growth, increase disease and stress tolerance, and reduce pathogens. In order to develop these new practices, we must gain an understanding of turfgrass microbial communities and how they are affected by management practices. We characterized bacteria, fungi, and nematodes on three golf courses: one organic, one with reduced inputs, and one conventional. We took samples from three management areas on each course representing different management intensities (roughs, fairways, and …

Engineering Novel Detection And Treatment Strategies For Bacterial Therapy Of Cancer, Jan T. Panteli

Doctoral Dissertations

Finding and treating cancer is difficult due to limited sensitivity and specificity of current detection and treatment strategies. Many chemotherapeutic drugs are small molecules that are limited by diffusion, making it difficult to reach cancer sites requiring high doses that lead to systemic toxicity and off-target effects. Tomographic detection techniques, like PET, MRI and CT, are good at identifying macroscopic lesions in the body but are limited in their ability to detect microscopic lesions. Biomarker detection strategies are extremely sensitive and able to identify ng/ml concentrations of protein, but are poor at discriminating between healthy and disease state levels due …

Bacterial Toxicity Of Oxide Nanoparticles And Their Effects On Bacterial Surface Biomolecules, Wei Jiang

Open Access Dissertations

Toxicity of nano-scaled Al2O3, SiO2, TiO2 and ZnO to bacteria (Bacillus subtilis, Escherichia coli and Pseudomonas fluorescens) was examined and compared to that of their respective bulk (micro-scaled) counterparts. All nanoparticles (NPs) but TiO2 showed higher toxicity than their bulk counterparts. Toxicity of released metal ions was differentiated from that of the oxide particles. ZnO was the most toxic among the three NPs, causing 100% mortality to the three tested bacteria. TEM images showed attachment of NPs to the bacteria, suggesting that the toxicity was affected by bacterial attachment.

The effects of oxide NPs on bacteria cells and bacterial surface …