Microbiology Commons^™

Genetic Screen For Regulators Of Pol4, A Dna Repair Polymerase In Saccharomyces Cerevisiae, Pasang Dolma Sherpa

Theses and Dissertations

In Saccharomyces cerevisiae, DNA polymerase 4 (POL4) is the beta repair polymerase, the product of the POL4 gene, and is involved in base excision repair (BER) and microhomology-mediated end joining (MMEJ). Despite its involvement in these repair pathways and its conservation across eukaryotes, deletion of POL4 shows no detectable phenotype under standard laboratory conditions. I hypothesized that unknown genes act as backup systems, providing redundant activities that allow pol4Δ cells to survive. To test this, a genetic screen for mutants showing synthetic lethality with pol4Δ was implemented. I constructed a strain lacking the POL4 gene and carrying …

The Impact Of Glucose Exposure On Organic Cation Transporters (Oct) Function In A Corneal Epithelial Cell Line, David A. Garza

Theses & Dissertations

Organic cation transporters (OCT) facilitate the movement of neurochemicals and medications, contribute to the removal of cellular waste, and facilitate neuronal communication. Three subtypes of these high-capacity, low-affinity transporters (OCT1, OCT2, and OCT3) are differently expressed in peripheral tissues such as the liver, kidneys, as well as the central nervous system. OCTs are also expressed in ocular tissues. Chronic hyperglycemia is a complication commonly associated with diabetes that can lead to ocular diseases such as diabetic keratopathy, which affects the cornea. In this project, we expose immortalized human corneal epithelial cells (HCE-S) to various glucose concentrations to study their impact …

Validation Of A Real-Time Pcr Laboratory Developed Test For The Detection Of Pneumocystis Jirovecii In Bronchoalveolar Lavage Samples, Caitlyn Moon

Honors Scholar Theses

Pneumocystis jirovecii is an opportunistic, host-obligate fungal pathogen responsible for causing P. jirovecii pneumonia (PJP) in humans. PJP is treated with trimethoprim-sulfamethoxazole (TMP-SMX), a strong broad-spectrum antibiotic that carries risk of renal and hepatotoxicity. Therefore, rapid diagnosis or exclusion of PJP is crucial to ensure PJP-positive patients receive effective treatment and PJP-negative patients avoid unnecessary antibiotics. Unlike most microorganisms, P. jirovecii cannot be cultured, presenting complications for the typical methods of diagnosing fungal pathogens. Historically, the gold standard for diagnosis was direct microscopic examination with stains such as methenamine silver and calcofluor white, but this diagnostic technique requires highly skilled …

Changing Dynamics Of Microbial Communities At A Hydrocarbon Contaminated Site And Methanogenic Interactions With Iron Minerals, Jakob Sell

Masters Theses

Microbial life is not only ubiquitous but also plays a critical role in the geochemical makeup of our planet. While researchers have long studied easily culturable aerobic microbes, the influence of anaerobic life on soils and mineral cycling is less understood. Notably, knowledge about interactions between anaerobic microbes and inorganic elements such as iron has progressed with the discovery that magnetite influences how anaerobic archaea carry out methanogenesis. To better understand anaerobic bacterial metabolisms and how they interact with minerals and subsurface hydrocarbons, we extracted 16S rRNA genes from two adjacent subsurface petroleum plumes, sequenced the DNA, and analyzed it …

A Tractable Nematode Model For The Emerging Fungal Pathogen, Candida Auris, Melissa Martinez

Dissertations and Theses (Open Access)

Candida auris is an emerging multidrug-resistant opportunistic fungal pathogen. It

poses a significant threat to immunocompromised patients and those with prolonged stays in

healthcare facilities. C. auris spreads rapidly in health care facilities through enhanced

colonization of skin and abiotic surfaces. The genetic factors contributing to this ability and to

the virulence, drug resistance, and stress-tolerant nature of C. auris are largely unknown.

Current animal models of C. auris infections are not well standardized and report conflicting

results. Additional animal models of virulence are needed, especially those amenable to high-

throughput analysis. The nematode Caenorhabditis elegans has been validated as …

Characterization Of Staphylococcus Aureus Clinical Isolates From Deep Musculoskeletal Infections For An In Vitro Biofilm Model For Phage Therapy, Raquel C. Luna

Dissertations and Theses (Open Access)

The rise of multidrug-resistant (MDR) infections is a growing concern in the treatment of deep musculoskeletal infections (MSKIs) and orthopaedic device-related infections (ODRIs). Staphylococcus aureus is responsible for about one third of these infections due to its strong biofilm forming capacity, presence on human skin, and the large number of antibiotic-resistant variants. Bacteriophage (phage) therapy is now being considered as a treatment for MDR MSKIs/ODRIs. We are investigating the use of phage encapsulated into or adsorbed onto biodegradable microspheres for local delivery to the site of these MDR infections. We determined that the anti-staphylococcal phage K effectively lyses the S. …

Characterization Of The Antiviral Activity Of Mouse Schlafen Proteins, Wendy Paola Alvarez Leal

Open Access Theses & Dissertations

The goal of this work was to characterize the anti-viral role of mouse Schlafen8 and Schlafen9. Expression of these proteins is induced by type I interferon in response to viral infections. Importantly, their human functional orthologs exhibit anti-viral functions, and the structural element required for these activities are conserved in Schlafen8 and Schlafen9. Therefore, it is expected that these proteins will also impair viral replication. An important limitation in this field is that the anti-viral function of human SLFNs has been evidenced only in in vitro cellular models. No informative mutations have been described in human populations so far. Thus, …

Donor And Recipient Microbiome Interactions Determine Functional Outcomes Of Fecal Microbiota Transplantation For Clostridioides Difficile Infection, Sophie A. Millard

All Dissertations

Clostridioides difficile is a major cause of healthcare-associated diarrhea. Infection typically occurs after antibiotic treatment disrupts the gut microbiome, allowing C. difficile to colonize and produce toxins leading to disease (CDI). Standard antibiotic treatments can resolve acute infection but frequently lead to recurrence, highlighting the need for alternative strategies that restore the gut ecosystem. Fecal microbiota transplantation (FMT), the transfer of stool from a healthy donor, has emerged as an effective therapy for recurrent C. difficile infection (rCDI). Understanding the mechanisms underlying the success of FMT for treatment of rCDI could lead to safer, more defined microbial mixtures.

This dissertation …

Degradation And Metabolic Analysis Of Synthesized Quorum-Sensing Signals By The Model Eukaryote Chlamydomonas Reinhardtii, Adam Josef Bach

Theses and Dissertations

Bacterial organisms can “talk” with one another on a colony-wide scale using a communication system known as quorum-sensing (QS) via chemical signals. These signals are produced at a basal rate throughout the life of the bacteria and are classified as autoinducers, which diffuse across the cell membrane. Once the critical concentration is reached, they begin to diffuse back into the cell and initiate a phenotypic change. As these signals exist outside the membrane of their producer, they are also accessible to other organisms. It has already been noted that some plant species uptake these compounds through their roots and break …

Understanding The Role Of Toxs As A Bile Sensing Component Of The Toxrs Virulence Regulatory System In Vibrio Spp., Minje Kim

Dartmouth College Ph.D Dissertations

Pathogenic Vibrio bacteria, such as V. cholerae and V. parahaemolyticus, colonize in the human small intestine to cause severe gastrointestinal disease. During the infection, these bacteria encounter bile salts, antimicrobial cholesterol metabolites secreted into the intestine. Pathogenic Vibrio species have evolved to utilize bile salts as signals to regulate virulence. The signaling depends in part on ToxRS, a conserved co-component transmembrane transcription regulator. ToxRS consists of the transcription factor ToxR and its membrane-tethered binding partner ToxS. ToxS is required for ToxR stability and full transcriptional activity. Although bile salts are known to influence ToxRS-dependent virulence gene expression, the molecular …

Descriptive Comparative Analysis Of Bacteriophage And Auxiliary Metabolic Gene Diversity In Marine And Wastewater Environments, Andrew Crawford

Undergraduate University Honors Capstones

Bacteriophages (phages) are viruses that infect bacteria and play a role in bacterial evolution. Through horizontal gene transfer (HGT), phages integrate parts of their genomes into their bacterial hosts. While the molecular mechanisms of this process are well understood, the evolutionary impacts remain less certain. One evolutionary explanation is that auxiliary metabolic genes (AMGs) modulate host gene expression to favor phage reproduction or improve the host’s survival. AMG can persist in hosts' or phages' genomes across future generations, altering their contributions to the ecosystem and posing a concern for human and environmental health. The objective of this project is to …

An Investigation Of Entamoeba Histolytica Chitin Synthases And Other Encystation Factors In Early Encystation, John N. Talledo

All Theses

Entamoeba histolytica is a food-and-water-borne human protozoan pathogen that relies on a two-stage life cycle for successful proliferation outside of the host. E. histolytica exists in its dormant cyst form while outside of its host and converts to the motile trophozoite (amoeba) form in the small intestine. The trophozoites colonize the large intestine to cause symptomatic disease in ~10% of infections, resulting in ~100 million cases of amoebic dysentery each year. Trophozoites can convert back to the cyst form, which is shed in feces to continue the infection cycle.

Currently, there is a lack of understanding of the exact molecular …

Investigating The Synthesis And Antibacterial Activity Of A Main-Chain Cationic Polymer, Emily Smith

Honors Theses

Antibiotic resistance is a growing threat to public health. Due to the overuse of antibiotics in agriculture, the overprescription of antibiotics, and the misuse of antibiotics by patients, antibiotic resistance is spreading at an alarming rate. As antibiotic resistance spreads, the antibiotics we depend on to treat common infections become ineffective. Therefore, it is imperative to develop novel treatments for infections that do not increase the prevalence of antibiotic resistance. One treatment being explored is the use of synthetic polymers with antibacterial properties. Synthetic polymers are advantageous because they can be synthesized from widely available precursors and their structure is …

Analysis Of Bacterial Virulence And Resistance In A Model Of Recurrent Uti, Rachel L. Proper

Undergraduate Honors Theses

Urinary tract infection (UTI) is a major women’s health issue, affecting approximately 40% of women during their lifetime. Formation of a biofilm by UTI-causing bacteria poses a major challenge in treatment, as the protection provided by the biofilm hinders both host defenses and antibiotic treatment. Failure to eradicate these infections causes them to become recurrent (rUTI). Previous research indicates that rUTI may be comorbid with a mutation in the SERPINE1 gene, which causes fibrin to accumulate within the body. Literature also indicates that fibrin provides a better scaffold for bacterial attachment and, subsequently, biofilm formation. In this work it was …

Microarray Analysis Of Human Abdominal Aortic Aneurysm With Emphasis On Cardiovascular Genes Revealed Differentially Expressed Genes, Song Lu, Li Ping Li, John V. White, Xiaoying Zhang, Ifeyinwa Nwaneshiudu, Adaobi Nwaneshiudu, Nectaria Ntaoula, John Gaughan, Dimitri S. Monos, Wan-Lu Lin, Charalambos C. Solomides, Emilia L. Oleszak, Chris D. Platsoucas

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

BACKGROUND/AIM: We examined gene expression profiles in abdominal aortic aneurysm (AAA) lesions vs. normal aortas by cDNA microarray and real-time quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR).

MATERIALS AND METHODS: Phosphorus (³²P)-labeled cDNA from AAA specimens (mean AAA size 6.65 cm) and normal aortas were hybridized with a 588-gene microarray primarily of the cardiovascular system. The results were validated by qRT-PCR.

RESULTS: A total of 35 out of the 588 genes were differentially expressed, with either log₂ ratio of AAAs/controls ≥1 (upregulated; 20 genes) or ≤-1 (downregulated; 15 genes) in AAA lesions vs. normal aorta, and …

Garlic Breath And Beads Of Sweat: The Effects Of Garlic And Chili Peppers On The Growth And Ph Of Streptococcus Mutans., Lena Pitzer

Honors Projects

The purpose of this project is to provide a quantifiable and repeatable measurement of how the presence of garlic and chili pepper during the growth of a carious dentine strain of Streptococcus mutans affects the growth of the strain and surrounding pH level.This research project examines two possible alternative agents, garlic and chili peppers, due to the known antimicrobial effects of their active components, allicin and capsaicin. Prepared garlic and chili extracts were used during disk diffusion to assess and measure the zones of inhibition. The anticipated results are that there will be inhibition present with both the garlic and …

Beyond Attachment: Endolysosomal Determinants Of New World Arenavirus Entry And Fusion, Cassandra E. Thompson

Arts & Sciences Graduate Student Theses and Dissertations

Arenaviruses are enveloped, negative-sense RNA viruses maintained in rodent reservoirs and transmitted to humans through exposure to aerosolized rodent excreta, where they can cause severe and sometimes fatal disease. The geographic distribution of arenaviruses reflects the range of their rodent reservoirs, resulting in the classification into Old World and New World groups, with New World arenaviruses further subdivided into clades A–D. Viral entry is the first step of infection and a major determinant of host range, tissue tropism, and zoonotic potential. Further, viral entry is a tightly regulated, multistep process that couple’s receptor engagement with intracellular trafficking and pH-dependent membrane …

Multi-Strain Assessment Of Quorum Sensing In Chlamydomonas Reinhardtii, Jacqueline Krueger

Ocean Engineering and Marine Sciences Student Publications

Quorum sensing (QS) allows microbial populations to coordinate phenotypic switching, selecting for specific behaviors that are only beneficial when expressed at high cell densities (HCD, >106 cells/mL) rather than low cell densities (LCD, < 106 cells/mL)

Investigating The Intra-Host Relationships Of The Microbiota At Multiple Sites, Ariel Jose Hernandez-Leyva

Arts & Sciences Graduate Student Theses and Dissertations

The human body hosts an abundance of microbial communities that, through complex and multi-faceted interactions, interface with host physiology and help govern health and disease. Many of these interactions are beneficial and help maintain host homeostasis or contribute to the metabolic capacity of the host. Some are detrimental, potentially inducing or further exacerbating disease. All are complicated by the vast diversity of microbes that comprise the microbiota at any given site. Identifying microbes or microbial functions that influence health and disease for study and potential use as biomarkers is tantalizing, though difficult. The vast diversity of potential community compositions, redundancy …

Elasmobranchii Oral Microbiome Diversity Within Southeast Florida: Implications Of Shark Zoonotic And Antibiotic Resistant Pathogens Associated With Human Interactions, Amanda Finnefrock

All HCAS Student Capstones, Theses, and Dissertations

Sharks are incredibly common in the Atlantic and Gulf waters surrounding Florida. Despite the prevalent nature of these animals, little research exists on the composition of their microbial communities (or “microbiomes”). Here we aim to determine the microbial makeup of the oral environment of sharks so that we can determine if there are any bacteria of clinical significance to humans, such as being antibiotic-resistant or having elevated capability for pathogenicity. A total of 41 samples were collected from the mouths and teeth of bull, Caribbean reef, great hammerhead, lemon, nurse, and sandbar sharks caught in Southeast Florida using sterile swabs …