Life Sciences Commons^™

Interactive Effects Of Sublethal Concentrations Of Fracking Biocides And Abandoned Mine Drainage On Amphipod Behavior, Kelly Lenhart

Electronic Theses and Dissertations

This thesis examined the sublethal effects of three pollutants, namely glutaraldehyde, 4,4-dimethyloxazolidine (DMO), and abandoned mine drainage (AMD), on amphipods. The primary objective was to investigate their combined effects on amphipods. The three pollutants, despite having the potential to combine in the environment, have not been studied to determine their potential for detrimental interactive effects which could result in unexpected environmental damage.

The research employed a series of experimental setups involving controlled exposure of amphipods to varying, putatively sublethal, concentrations of the chemicals of interest. Subsequently, effects were assessed via both behavioral and feeding assessments. To facilitate this analysis, novel …

Analyzing Pseudomonas Aeruginosa With Bacteriophage Tags Using Photoacoustic Flow Cytometry, Jennifer C. Schinke

Electronic Theses and Dissertations

The number of daily bacterial infections is climbing and the CDC explains that this is due to the antibiotic-resistant threat in the United States. Finding a faster way of bacterial identification is necessary as it currently takes 1-4 days for a medical lab to culture and identify bacteria. Photoacoustic flow cytometry (PAFC) can be used as an alternative method resulting in swift identification within an hour (Edgar, 2019). Pseudomonas aeruginosa, cell line PA01, will be coated in up to a few hundred red dyed phages making it detectible by the photoacoustic flow cytometry system. Bacteriophages (phages) are viruses that …

Assessing The Appropriateness Of Free-Roaming Cat Management Strategies Based On Resident Perspectives In A Small Pennsylvania Municipality, Brianna Marks

Electronic Theses and Dissertations

The popularity and mismanagement of free-roaming cats has resulted in invasive populations responsible for over-predation of birds and small mammals and increased disease transmission to humans and animals (Bies, 2014; Crowley et al., 2020). Management effectiveness fluctuates between communities due to varying opinions of free-roaming cats by governments and the public, which can influence management development and success (Deak et al., 2019). This research assesses the relationship between community perspectives of free-roaming cats and management. A survey of residents in a small Pennsylvania municipality was conducted with questions pertaining to free-roaming cats and management preferences in the community. Over 50% …

Detecting Native Freshwater Mussels In Pennsylvania Waterways: Comparison & Validation Of Environmental Dna Methods, Meredith Bennett

Electronic Theses and Dissertations

North America is home to approximately one third of the world's freshwater mussel species. They are highly imperiled organisms due to habitat destruction and invasive species. Traditional surveys rely on visual identification of mussels, but individuals tend to be rare and difficult to identify. An alternative method is to extract environmental DNA (eDNA) from water samples, which has advantages over traditional sampling, including less sampling effort and fewer hazards to researchers and organisms. We conducted a review of the two main eDNA approaches: single-species detection and metabarcoding. We also developed and validated metabarcoding primers for the detection of native mussels. …

Quantum Computations And Molecular Dynamics Simulations: From The Fundamentals Of Antimicrobial Resistance To Neurological Diseases, Angel Tamez

Electronic Theses and Dissertations

Biophysical phenomena are modeled using a combination of quantum and classical methods to interpret and supplement three distinct and diverse problems in this dissertation. In the first project, decarboxylation reactions are ubiquitous across chemical and biological disciplines, yet the origin of non-catalytic solvent effects remains elusive. Specific solvent structure and energetics have not been well described for the monoanion of malonate, nor corrected from the gas-phase charge-assisted intramolecular hydrogen bond model known as “pseudochair”. In the aqueous phase, a low-lying energy conformer known as the “orthogonal conformation” is computed to be preferred by a three-water cluster of hydrogen bonding over …

Neural Stem Cells: Age-Dependent Outcomes During Viral Infections In The Central Nervous System, Manisha N. Chandwani

Electronic Theses and Dissertations

Viral infections in the central nervous system (CNS) are associated with neurodevelopmental and neurobehavioral deficits. The outcomes of viral infections can be driven by damage and death of neurons. Neural stem cells (NSCs) play key roles in neurodevelopment, repair, and physiological brain function. During a viral infection, NSC activity can disturbed by direct infection of NSCs by the virus or by anti-viral immune response. Here, we aimed to assess whether the anti-viral immune response can impact NSC activity during an immunocompetent response in the adult brain. We utilized a transgenic mouse model of Measles virus infection where only the CNS …

Novel Signal Sequences And Fusion Partners For Paratransgenesis In Asaia, Christina Grogan

Electronic Theses and Dissertations

Mosquitoes transmit many pathogens that cause human disease. One such disease, malaria, is caused by parasites in the genus Plasmodium, infecting over 200 million people and killing over 600,000 per year. Current strategies to control vector-transmitted diseases are increasingly undermined by mosquito and pathogen resistance. Research has turned to additional and novel methods of control, such as altering the microbiota of the vectors. In this method, called paratransgenesis, symbiotic bacteria are genetically modified to affect the mosquito’s phenotype by engineering them to deliver antiplasmodial molecules into the midgut to kill parasites. These molecules must be released by the …

Investigating Host Defenses Of North American Salamanders Against The Recently Emerged Chytrid Pathogen, Batrachochytrium Salamandrivorans, Kenzie Pereira

Electronic Theses and Dissertations

A recently emerged chytrid fungal pathogen, Batrachochytrium salamandrivorans (Bsal) threatens salamander biodiversity. Bsal susceptibility varies between and within salamander species, but little is known about the mechanisms underlying these differences. Susceptibility is likely influenced by numerous interacting factors, but my dissertation studied the role of host immune responses.

My first aim investigated between species differences by studying the bioactive properties of salamander skin peptides against Bsal and the related pathogen, B. dendrobatidis (Bd). Skin peptides were collected from five salamander species, used for in vitro assays, and analyzed by RP-HPLC. While skin peptides from one …

Identification And Characterization Of Genetic Elements That Regulate A C-Di-Gmp Mediated Multicellular Trait In Pseudomonas Fluorescens, Collin Kessler

Electronic Theses and Dissertations

Microbial communities contain densely packed cells where competition for space and resources are fierce. These communities are generally referred to as biofilms and provide advantages to individual cells against immunological and antimicrobial intervention, dehydration, and predation. High intracellular pools of cyclic diguanylate monophosphate (c-di-GMP) cause cells to aggregate during biofilm formation through the production of diverse extracellular polymers. Genes that encode c-di-GMP catalytic enzymes are commonly mutated during chronic infections where opportunists display enhanced resistance to phagocytosis and antibiotics. Our lab uses an emergent multicellular trait in the model organism Pseudomonas fluorescens Pf0-1 to study the emergence of c-di-GMP mutations …

A Bioaffinity Self-Assembling Gel For Localized Delivery Of Immunotherapeutics, Ngoc Pham

Electronic Theses and Dissertations

Renal cell carcinoma (RCC) is a type of cancer in which malignant cells develop in the kidney tubules. Immune checkpoint inhibitors (ICI), including anti-PD-1 antibodies (aPD1), as monotherapies or in combination settings have advanced to the front-line treatment for patients diagnosed with RCC. Systemic infusion of ICI may render subtherapeutic concentrations in the lesions and increase risks of immune-related adverse events. To overcome these limitations, locoregional drug delivery has gained substantial interests as a modality in immuno-oncology. However, conventional intratumoral injections may be ineffective because the high interstitial fluid pressure in tumors resists drug retention and results in rapid local …

Sex-Specific Effects Of Chaperone And Glial Defenses On Experimental Lewy Body Disease, Tarun Bhatia

Electronic Theses and Dissertations

Lewy body disorders are a group of neurodegenerative conditions characterized by the pathological misfolding and aggregation of the abundant protein, α-synuclein. The most common Lewy body disorders are Parkinson’s disease and dementia with Lewy bodies. Apart from ageing, male sex is a major risk factor for Lewy body disorders, as men are at ~1.5-fold higher risk for these diseases than women. Yet, preclinical studies on Lewy body disorders rarely examine sex as a biological variable, and the mechanisms underlying sex-skewedness in disease risk remain undetermined.

Here, we developed a sex-stratified model of Lewy body disorders by exposing primary neurons harvested …

Evaluation And Application Of Crispr-Cas12a-Based Systems For Amphibian Detection From Dietary And Environmental Dna, Macie Chess

Electronic Theses and Dissertations

Current methods for monitoring biodiversity are hindered by detection efficiencies or rely on expensive laboratory equipment. The goal of this study was to evaluate the application of a CRISPR-Cas12a-based system for the species-specific detection of amphibians from dietary and environmental DNA samples. A Cas-12a-based platform was designed and analyzed for the fluorescent detection of three salamander species; Desmognathus fuscus, D. ochropheaus, and Erycia bislineata. This system was applied to the identification of salamander DNA extracted from the fecal sacs of nestling Louisiana Waterthrush (Parkesia monticola), amplified with PCR primers targeting the CYT B mtDNA gene, and compared to DNA metabarcoding. …

Evaluating Threats To The Water Quality Of Little Buffalo Creek And Sarver Run Through Water Quality Analysis And Backpack Electrofishing Surveys, Riley Williams

Electronic Theses and Dissertations

Currently, 64.8% and 52.2% of Little Buffalo Creek and Sarver Run stream kilometers are impaired, respectively. Impairment sources to Little Buffalo Creek include agriculture, riparian deforestation, on-site wastewater, and urban runoff. Impairment sources to Sarver Run are unknown. Our goal was to understand sources of impairment through water quality analysis and fish surveys. Water samples were collected monthly and analyzed for cation and anion concentrations. Mass ratio analysis comparing Br/SO₄ and Mg/Li, Ca/Mg and Ca/Sr, Mg/Na and SO₄/Cl, and SO_4/Cl and Mg/Li suggests that mine drainage and conventional oil and gas may be impacting water …

A Review Of Current Methods In Avian Dietary Analysis And Their Integrated Application To Characterize The Trophic Niche Of Louisiana Waterthrush (Parkesia Motacilla)., Brandon Hoenig

Electronic Theses and Dissertations

Characterizing a species’ dietary composition presents an avenue to understand many facets of its ecological niche and can provide essential information for the species’ long-term conservation. To date, the vast majority of diet studies have relied on direct identification of prey during foraging observations or from diet samples to characterize the dietary habits of birds. However, advancements in laboratory-based approaches have revolutionized the field of trophic ecology by allowing researchers to indirectly infer dietary habits with higher resolution across greater time scales. Here, I apply two of these laboratory-based techniques, namely DNA metabarcoding and stable isotope analysis, to characterize the …

Analysis Of Water Quality And Fish Communities Of Crooked Creek, Tributary To The Allegheny River In Southwestern Pennsylvania, Katherine Stupar

Electronic Theses and Dissertations

Crooked Creek is a tributary of the Allegheny River with a drainage area of 292 square miles. It has been historically affected by mine drainage pollution. Extensive cleanup and monitoring efforts by the Commonwealth of Pennsylvania and the U.S. Army Corps of Engineers (USACE) have taken place over the last four decades. Unconventional oil extraction, or fracking, presents an emerging threat to the water quality of this stream and the health of its fish communities. Here we perform an analysis of the Crooked Creek watershed utilizing backpack electrofishing, eDNA metabarcoding, and various methods of water quality analysis. Seven sites on …

Real-Time Monitoring Of Cell Death Progress Using Capacitance Spectroscopy, Suyang Wu

Electronic Theses and Dissertations

Biologics, including the monoclonal antibody (mAb), has experienced rapid development in the last decade. However, the price of biologics is often prohibitively high because of the low process efficiency. Delaying the inevitable cell death improves the productivity of upstream bioprocessing, whose success relies on monitoring the cell death onset that indicates the timing for preventive actions.

This study proposes to develop a real-time monitoring model that quantifies the dying cell percentage in lab-scale bioreactors using capacitance spectroscopy. The capacitance spectroscopy contains cell death-related information due to various physical properties changes during the cell death process, e.g., cytoplasmic conductivity change. The …

Understanding Snow Leopard (Panthera Uncia) Population Structure, Diet, And Human-Wildlife Dimensions Using Noninvasive Genetic Approaches, Charlotte Hacker

Electronic Theses and Dissertations

Snow leopards (Panthera uncia) are a large cat endemic to the mountains of Central Asia. Despite their status as a flagship species, there are numerous knowledge gaps surrounding their population status, dietary ecology, and relationship with humans. Such knowledge gaps are prohibitive for effective conservation action for the species. Snow leopards are threatened by habitat loss, decreased prey availability, retaliatory killings, and large-scale negative impacts associated with climate change. In order to help bridge these gaps, noninvasive genetic methodologies were deployed in this work across several disciplines. Population genetic metrics were garnered at local, regional, and range-wide scales, …

Analysis Of Spore Shape Determination In Streptomyces, Ning Sun

Electronic Theses and Dissertations

Streptomycetes are Gram-positive, soil-dwelling bacteria that possess a complex life cycle with the alternation of vegetative mycelium, aerial mycelium, and spores. Streptomyces. coelicolor spore maturation is a complex process that involves spore shape metamorphosis from cylindrical pre-spores into ellipsoid spores, but the details of this process have remained enigmatic. Previously, our lab identified a novel gene ssdA that might play a role in spore shape determination using a transposon-based insertion mutagenesis in S. coelicolor. In this study, I isolated a S. coelicolor ssdA-null mutant that showed increased colony hydrophobicity and misshapen spores in sizes and shapes, confirming the …

Cgrp-Driven Lateralization Of Pain Modulation By The Central Amygdala, Heather Allen

Electronic Theses and Dissertations

The central amygdala (CeA) is a bilateral, mid limbic brain region involved in the processing of pain and emotion. Interestingly, the left and right CeA are functionally lateralized in the context of bladder pain, with the left and right CeA having opposing effects on bladder pain-like behavior. However, the specific mechanisms driving these divergent functions are unknown. Nociceptive signals from the bladder are relayed to the CeA via projection neurons in the parabrachial nucleus (PBN) that express high levels of calcitonin gene-related peptide (CGRP). In these studies, we investigated the influence of PBN CGRP signaling in the left and right …

Identification Of Fluoxetine-Sert Interactions And Apo-Sert Studies Via Crosslinking Mass Spectrometry, Elizabeth Castellano

Electronic Theses and Dissertations

The serotonin transporter (SERT) is a member of the neurotransmitter sodium symporter family of transporters. SERT controls the magnitude and duration of serotonergic neurotransmission by facilitating the reuptake of serotonin back into the pre-synaptic neuron and is thus a target for antidepressants. Selective serotonin re-uptake inhibitors (SSRIs) such as fluoxetine, are commonly prescribed to treat depression. SSRIs act by blocking reuptake and prolonging serotonin signaling. However, significant problems regarding selectivity and mechanisms of action of these drugs remain unresolved. The structures of SERT and related transporters have been determined and serve as useful structural models. However, they are typically mutated …

Characterizing The Dormancy And Repair Of A Circumneutral Passive Remediation System Receiving Iron And Sulfate-Rich Amd, Garrett Struble

Electronic Theses and Dissertations

The Wingfield Pines remediation system is a ~20-acre circumneutral, gravity-fed system constructed in 2009 to passively remediate Fe from an ~2000 gallon/minute effluent. In 2017, a fracture to the system’s underground cavern caused the AMD effluent to bypass the system, flowing directly into Chartiers Creek and leaving the system decommissioned between 2017-2019. Soil slurry and water samples were collected in the weeks prior to the system’s reinstalment in September 2019 through January 2020 in order to determine the effect of the repair on remediation efficiency and microbial community composition. This study acted as the first to describe the temporary decommissioning …

Rna G-Quadruplex Structures: Functions In Noncoding Rnas And Disease, Emily Benner

Electronic Theses and Dissertations

Paraspeckles are a relatively newly identified subnuclear body whose function is still largely unknown. They are membraneless organelles found within mammalian nuclei originally identified as being comprised of different paraspeckle proteins (PSPs), and later proven to also contain RNA. This study investigates the long non-coding RNA nuclear enriched abundant transcript 1 (NEAT1), which has been found to be indispensable in the formation of paraspeckles, with each containing up to 50 NEAT1 molecules. NEAT1 has two isoforms, the shorter NEAT1_1, and the longer NEAT1_2. The focus of this work will be on the NEAT1_2, which acts as a scaffold in the …

Antiparasitic Peptide From The Ocean: Discovery, Synthesis And Structure-Activity Relationship Study, Kh Tanvir Ahmed

Electronic Theses and Dissertations

Millions of people die every year because of infectious diseases, and malaria is among the top five of the deadliest infectious disease. In 2018, malaria took more than four hundred thousand lives, and more than half of them are children under five years of age. Most malaria-affected parts of the world are also the home of the most underprivileged people. Seemingly, antimalarial drug discovery never achieved the attraction that it requires. Like malaria, another infectious disease that is not extensively explored by drug discovery campaigns is American trypanosomiasis or Chagas disease. More than a hundred years have passed since discovering …

Identifying The Effects Naturally Forming Bacterial Communities Have On The Efficiency Of Passive Remediation Systems Built To Treat Abandoned Coal Mine Drainage, Michelle M. Valkanas

Electronic Theses and Dissertations

Abandoned mine drainage (AMD) has resulted in worldwide devastation. Passive remediation systems (PRS) are a cost-effective option to treat AMD. While the geochemistry (abiotic) of these systems is well understood, little work has been done to characterize the microbial communities (biotic) that naturally form within these systems. The goal of this dissertation was to identify the effects that bacterial communities have on the efficiency of PRSs. Four passive remediation sites built to treat coal mine drainage were studied: two that treat acidic discharges (Boyce and Middle Branch) and two that treat circumneutral discharges (Wingfield Pines and Lowber). Survey studies suggested …

Rna Structure And Function: Biological Relevance In Neurodegenerative And Infectious Disease Pathogenesis, Joshua Imperatore

Electronic Theses and Dissertations

The studies outlined in this dissertation encompass a broad focus, relating to the pathogenesis of neurological disorders such as Alzheimer’s disease, fragile X syndrome, and amyotrophic lateral sclerosis, as well as a novel infectious disease, COVID-19. However, all four studies detailed within this dissertation contain similar elements, namely, the coordinated role of RNA structure and function in essential molecular mechanisms. The results of these investigations allow us to speculate on various disease- related mechanisms, including dysregulated microRNA processing pathways, nucleocytoplasmic shuttling via distinct RNA-protein interactions, and fine-tuned molecular switches controlling genomic dimerization and viral hijacking of host cellular microRNAs. Taken …

Isolation, Structure Elucidation, And Synthesis Of Natural Products From Marine Cyanobacteria, Keren Solomon

Electronic Theses and Dissertations

This thesis describes the isolation, structure elucidation, and synthesis of natural products from marine cyanobacteria. A crude extract from a cyanobacterium collected in Curacao showed selective affinity for the dopamine D₅ receptor in a screen against a panel of CNS receptors. Due to the high similarity of the D₅ and D₁ receptor, to date there are no known ligands that differentiate them. Attempts to purify the compound responsible for this affinity led to the isolation of the known compound caylobolide A. A second extract from a cyanobacterium collected in Panama underwent bioassay-guided fractionation and yielded the novel …

Differential Expression Of Rna In The Rat Peripheral Nervous System Following Nerve Injury And Treatment With Pain-Relieving Celecoxib-Loaded Nanomedicine, Andrea Stevens

Electronic Theses and Dissertations

The neuroinflammatory response to peripheral nerve injury is associated with chronic pain and significant changes in the expression profiles of RNAs in neurons, glia and infiltrating immune cells: a neuro-immune triad. Chronic constriction injury (CCI) of the rat sciatic nerve provides an opportunity to mimic neuropathic injury and quantitatively assess behavior and differential gene expression in individual animals. Macrophages that phagocytose intravenously injected nanoemulsion carrying the non-steroidal anti-inflammatory, NSAID, Celecoxib, naturally accumulate at the site of injury resulting in relief of CCI behavioral hyper-sensitivity. It is not known beyond the inhibition of cyclooxygenase-2 (COX-2) activity and the reduction in prostaglandin …

Evaluating The Biological Effects And Safety Of Exposure To Low, Putatively Safe Concentrations Of The Common Pesticide Chlorpyrifos Using An Anuran Model, Sara Mcclelland

Electronic Theses and Dissertations

In the US, over 1.1 billion pounds of pesticides are applied annually. Unfortunately, pesticides contaminate natural habitats affecting animal health. Chlorpyrifos (CPF) is an organophosphorous insecticide that contaminates surface waters. Few studies have assessed the impacts of CPF at low, ecologically realistic doses. I aimed to test the effects of low dose CPF exposures on biological parameters including body morphology, hormone levels, and brain development in Northern Leopard Frogs (Lithobates pipiens), a common vertebrate model. To determine if CPF exposure has direct impacts or indirectly impacts animals through the disruption of the food web, I examined CPF exposure …

Development Of A Pharmaceutical Tablet Authentication System Using Spectroscopic Techniques In Combination With Multivariate Chemometric Methods, Md Nayeem Hossain

Electronic Theses and Dissertations

The spread of falsified drugs is increasing worldwide. Currently, 10%-30% of drugs in the world are falsified. Unfortunately, the supply chain system of developing countries (from manufacturers to customers) is not well monitored and suffers the highest rates of fraudulent activities. Also, the rise of product procurement from the internet increases the chance of American consumers' exposure to poor quality drugs. To combat this horrendous activity, surveillance of pharmaceutical materials is required in the supply chain system. Spectroscopic techniques (e.g., Near-Infrared and Raman spectroscopies) can be a potential solution to authenticate samples in different locations; they are non-destructive, safe, rapid, …

Pilot Study Exploring The Effect Of Targeted Cox-2 Inhibition In Macrophages Responding To Neuronal Injury; Promoting Enhanced Axonal Regeneration, Alyssa Brauckmann

Electronic Theses and Dissertations

Celecoxib nanoemulsion (CXB-NE) has been developed as a macrophage targeted analgesics by Dr. Janjic and her team at Duquesne University, (Janjic et al, 2018; Liu et al, 2020; Saleem et al, 2019b; Vasudeva et al, 2014). The CXB-NE nanoemulsion carrying a Nonsteroidal Anti-inflammatory (NSAID) inhibitor of COX-2 activity result in a reduction in PGE2 expression in macrophages. Using CXB-NE in rats that have peripheral nerve injury constricting the sciatic nerve relieves hypersensitivity, a pain-like behavior. The treatment also decreases inflammation associated with this chronic constriction injury (Janjic et al, 2018; Saleem et al, 2019b; Stevens et al, 2019). In this …