Medicine and Health Sciences Commons^™

A Potential Klebsiella Bacteriocin With Efficacy Toward The Enterbacteriaceae Family, Kasey Barber

Undergraduate Honors Theses

Drug resistance is unfortunately becoming a prevalent issue in the course of patient treatment, ranging from chemotherapy resistance to antimicrobial resistance. The Centers for Disease Control and Prevention (CDC) estimated in 2016 that at least 23,000 people die every year in the United States from an infection with an antibiotic-resistant organism (Munita, et al, 2016). Carl Friedlander was the first scientist to describe Klebsiella pneumoniae in 1882 as an encapsulated bacillus after isolating the bacterium from the lungs of patients who had died from pneumonia (Ashurst and Dawson, 2022). Klebsiella pneumoniae is the type species for the Klebsiella genus and …

Nicotinamide Riboside And Beta-Hydroxybutyrate Activate Parallel Pathways For C. Elegans Lifespan Extension, Mckenzie Peters

Undergraduate Honors Theses

Supplementation with nicotinamide riboside (NR), a form of vitamin B3 and a precursor of nicotinamide adenine dinucleotide (NAD⁺) extends lifespan in the nematode C. elegans and delays aging-related pathologies in mammals. During aging, levels of NAD⁺ decline causing metabolic dysfunction and oxidative damage. Studies in C. elegans found that when NR was administered during larval development it induced the mitochondrial unfolded protein response (UPR^mt), which is frequently associated with lifespan extension. Both calorie restriction (CR) and ketogenic diets (KD) have been shown to extend lifespan, in part through increasing NAD⁺ and through increasing levels …

Early Detection Of Atypical Motor And Neurobehavior Of Infants At Risk Secondary To Opioid Exposure: A Prospective Study, Kara Boynewicz

Electronic Theses and Dissertations

Prenatal opioid exposure has been studied in relation to infants' medical outcomes. However, large gaps exist in the literature supporting early identification of atypical neurobehavior and motor development of infants with prenatal opioid exposure. The purpose of the study was to investigate whether prenatal opioid exposure has a negative influence on a newborn infant’s neurobehavior and motor development to aid in the early identification of potential delays. Using a prospective quasi experimental design, infants motor development using the Test of Infant Motor Performance (TIMP) and neurobehavior using the NICU Neonatal Network Scale (NNNS) was assessed on 58 infants in a …

Effects Of Nicotinamide Riboside And Beta-Hydroxybutyrate On C. Elegans Lifespan, Jeffery Peters

Undergraduate Honors Theses

The nicotinamide riboside (NR) form of vitamin B₃and the ketone body ß-hydroxybutyrate (BHB) are two of the most promising natural compounds yet identified for the treatment of aging and aging-related diseases. Forms of vitamin B₃are precursors for the synthesis of the coenzymes nicotinamide adenine dinucleotide (NAD(H)) and nicotinamide adenine dinucleotide phosphate (NADP(H)). In aged cells levels of NAD⁺decline, decreasing metabolism and decreasing activity of protective sirtuin protein deacetylases. In aged cells NR, but not more common forms of vitamin B3, boost NAD⁺levels. BHB is naturally produced by the body when individuals fast …

Ctrp3 And Alcoholic Liver Disease In Female Mice, Callie Root

Undergraduate Honors Theses

C1q TNF Related Protein 3 (CTRP3), is a cytokine that is primarily secreted from adipose tissue, which classifies it as an adipokine. Our previous research has shown that CTRP3 prevents alcoholic fatty liver disease (ALD) in male mice. However, even when accounting for confounding factors such as absolute and relative alcohol intake, females are more sensitive to the effects of consumption compared to male mice. Therefore, the goal of this project was to determine whether CTRP3 prevented ALD in female mice. Methods: Female wild type (WT) and female CTRP3 transgenic over expressing (Tg) mice were fed an ethanol containing liquid …

Isolation, Analysis, And Partial Characterization Of An Inhibitor Of Neisseria Gonorrhoeae, Natania Paul

Undergraduate Honors Theses

There is an emerging threat of Neisseria gonorrhoeae strains that are resistant to all antibiotics. Because of this, the purpose of this research is to isolate, analyze, and partially characterize a new inhibitor(s) of N. gonorrhoeae. Since there is an unknown molecule secreted by Candida albicans that inhibits N. gonorrhoeae, this molecule can be partially characterized using ¹H NMR Spectroscopy to assist in the development of a new antibiotic compound. It was hypothesized that quorum-sensing molecules, trans, trans- farnesol, tyrosol, phenylethyl alcohol, and tryptophol, could be possible candidates for the inhibitor. Because of this, ¹H …

The Study Of Alcoholic Liver Diseases, Seun Emmanuel Owoseni

Electronic Theses and Dissertations

Excessive alcohol consumption is the primary contributing factor in the development of alcoholic liver diseases (ALD). Nicotine contained in tobacco is a major addictive alkaloid, which enhances the effects of ALDs. The major enzyme involved in nicotine metabolism is cytochrome P450 2A5 (CYP2A5) which is produced in the liver. Alcohol can stimulate the CYP2A5 enzyme. We utilized cyp2a5-/- knockout mice in this research to examine the effects of CYP2A5.

The cyp2a5-/- mice and wild-type (WT) mice were fed liquid ethanol diet with or without nicotine to induce ALD. Nicotine enhancing effects on ALD were observed in WT mice but not …

Refrigerated Stability Of Diluted Cisatracurium, Rocuronium, And Vecuronium For Skin Testing After Perioperative Anaphylaxis, Kristen Dinsmore, Bethany Campbell, Timothy Archibald, Greg Mosier, Stacy Brown Phd, Alexei Gonzalez-Estrada Md

Appalachian Student Research Forum

Rationale: The purpose of this study is to investigate the stored stability of dilutions of neuromuscular blocking agents (NMBAs), namely cisatracurium, rocuronium, and vecuronium, for skin prick/intradermal testing.

Methods: Concentrations of NMBAs were monitored by liquid chromatography-mass spectrometry (LC-MS/MS) for a period of 14 days. Dilutions of NMBAs were prepared in saline by factors of 10x, 100x, 1,000x, 10,000x, and 100,000x as sensitivity of the assay allowed. Diluted drug products were stored in a laboratory refrigerator until sampling. On sampling days, aliquots of each dilution were removed and compared to a freshly prepared set of reference dilutions.

Results: The results …

Synthesis And In-Vitro Cell Viability/Cytotoxicity Studies Of Novel Pyrrolobenzodiazepine Derivatives, John M. Jarrett

Undergraduate Honors Theses

Pyrrolobenzodiazepines (PBDs) are a group of naturally occurring compounds that were discovered in the cultures of Streptomyces in the 1960s. Some natural PBDs discovered in these cultures, such as anthramycin and sibiromycin, were shown to possess a broad spectrum of anti-tumor activity. Since cancer is still a leading cause of death globally, the development of novel anti-proliferative derivatives of PBDs is essential for human welfare worldwide. Further synthesis and structure-activity relationship (SAR) studies of the parent natural products and their tetracyclic analogs will lead to the discovery of drug candidates. In this work, thirteen PBD analogues were synthesized using no …

Chat Expression In Chlamydia Muridarum-Infected Female Murine Genital Tract, Hallie Sartain

Undergraduate Honors Theses

Chlamydia trachomatis is the most prevalent agent of bacterial sexually transmitted infections in the world. However, a profuse number of cases are unreported, as the infection is often asymptomatic. Sequelae such as pelvic inflammatory disease, an increased risk of cervical cancer, premature birth, and perinatal infections in pregnant women can occur. Inflammation occurs in the body in response to infection or injury. Although inflammation can lead to some unwanted secondary effects, such as pain, it serves to return the body to homeostasis by restoring injured tissues and eliminating pathogens. One recently identified connection between the central nervous system and the …

The Effects Of Chronic Alcohol Consumption On Ovarian Function/ Morphology, Destiny Roberts

Undergraduate Honors Theses

Chronic alcohol (ethanol) consumption has been known to affect the major organs of the body and particularly the liver. However, the effects of chronic ethanol consumption on the female reproductive system remain relatively unstudied. A convenient way to study these effects is by analyzing laboratory mice that have been fed an ethanol diet for an extended period of time and comparing them to control mice. In this study, female mice were separated into control and ethanol fed groups. The mice were placed on their specified diets and observed over the course of six weeks. The mice were fed and weighed …

Sex Differences In The Kinetic Profiles Of D- And L- Methylphenidate In The Brains Of Adult Rats, J. Bentley, F. Snyder, Stacy D. Brown, R. W. Brown, Brooks B. Pond

ETSU Faculty Works

OBJECTIVE: Methylphenidate is commonly used in the treatment of Attention Deficit Hyperactivity Disorder and narcolepsy. Methylphenidate is administered as a racemic mixture of the d- and l- threo enantiomers; however, the d-enantiomer is primarily responsible for the pharmacologic activity. Previous studies of the behavioral effects of methylphenidate have highlighted sex differences in the responsiveness to the drug, namely an increased sensitivity of females to its stimulatory effects. These differences may be due to differences in the uptake, distribution, and elimination of methylphenidate from male and female brains. Therefore, we compared the pharmacokinetics of d- and l- threo methylphenidate in the …

Physiologically-Based Pharmacokinetic Model For Ertapenem, Whitney Forbes

Electronic Theses and Dissertations

Ertapenem is a carbapenem used to treat a wide range of bacterial infections. What sets ertapenem apart from other carbapenems is its longer half-life which implies it need only be administered once daily. We developed a physiologically-based pharmacokinetic model for the distribution of ertapenem within the body. In the model, parameters such as human body weight and height, age, organ volumes, blood flow rates, and partition coefficients of particular tissues are used to examine the absorption, distribution, metabolism, and excretion of ertapenem. The total and free blood concentrations found were then compared to experimental data. We then examined the sensitivity …

A Systematic Column Performance Comparison For The Confirmation Of Opioids Used In Pain Management By Lc-Ms, Derek Stallard

Undergraduate Honors Theses

In this study, three different chromatographic column chemistries (C18, Pentafluorophenyl (PFP), Hydrophilic Interaction Chromatography (HILIC)) were compared under optimal conditions to determine which stationary performed best in the separation and detection of a mixture of opioids using LC-MS. Furthermore, these stationary phases were examined in three different column technologies – traditional silica, porous shell, and porous polymer (PRP). The PRP column had the best peak shape for all 13 opioids and dominated for later-eluting compounds. In terms of column reproducibility, the Hamilton C18 column had the lowest %RSD values. The Kinetex HILIC produced the most theoretical plates and best resolution …

Random Mutagenesis Of The Aspergillus Oryzae Genome Results In Fungal Antibacterial Activity, Cory A. Leonard, Stacy D. Brown, James Russell Hayman

ETSU Faculty Works

Multidrug-resistant bacteria cause severe infections in hospitals and communities. Development of new drugs to combat resistant microorganisms is needed. Natural products of microbial origin are the source of most currently available antibiotics. We hypothesized that random mutagenesis of Aspergillus oryzae would result in secretion of antibacterial compounds. To address this hypothesis, we developed a screen to identify individual A. oryzae mutants that inhibit the growth of Methicillin-resistant Staphylococcus aureus (MRSA) in vitro. To randomly generate A. oryzae mutant strains, spores were treated with ethyl methanesulfonate (EMS). Over 3000 EMS-treated A. oryzae cultures were tested in the screen, and one isolate, …

A Mathematical Model For Antibiotic Resistance In A Hospital Setting With A Varying Population, Edward H. Snyder

Electronic Theses and Dissertations

Antibiotic-resistant bacteria(ARB) is causing increased health risk and cost to society. Mathematical models have been developed to study the transmission of resistant bacteria and the efficacy of preventive measures to slow its spread within a hospital setting. The majority of these models have assumed a constant total hospital population with the admission and discharge rates being equal throughout the duration. But a typical hospital population varies from day to day and season to season. In this thesis, we apply variable admission and discharge daily rates to existing deterministic and stochastic models which examine the transmission of single and dual resistant …

Stability Of Extemporaneously Prepared Lansoprazole Suspension At Two Temperatures, Jordan T. Morrison, Ralph A. Lugo, Jim C. Thigpen, Stacy D. Brown

ETSU Faculty Works

OBJECTIVE The purpose of this study was to examine the stability of a generic lansoprazole product in a 3 mg/mL sodium bicarbonate suspension under room temperature and refrigerated conditions. METHODS Lansoprazole suspensions (3 mg/mL) were prepared in triplicate using an 8.4% sodium bicarbonate vehicle for each storage condition (room temperature and refrigerated). During 1 month, samples from each replicate were periodically removed and analyzed for lansoprazole concentration by liquid chromatography–tandem mass spectrometry (LC-MS/MS). Each sample was spiked with 10 mg/L omeprazole to serve as the internal standard. A positive electrospray LC-MS/MS method was validated over the calibration range of 5 …

The Effects Of Nicotine Administration On Behavior And Markers Of Brain Plasticity In A Rodent Model Of Psychosis, Marla K. Perna

Electronic Theses and Dissertations

Schizophrenia affects about 1% of the population. A hallmark of the disorder is increased dopamine D2 receptor sensitivity in the brain. Studies have shown that schizophrenics smoke cigarettes at approximately 4 times the rate of the general population. It has been suggested that nicotine use is a form of self-medication for symptoms in schizophrenia. Smoking behaviors typically begin in adolescence. We assessed effects of nicotine on behavior and brain plasticity in an adolescent rodent model of schizophrenia with the goal of identifying targets for smoking cessation. Methods: Rats were neonatally treated with quinpirole (a D2/D3 agonist) or saline and sensitized …

The Fate Of Sulfamethazine In Sodium-Hypochlorite-Treated Drinking Water: Monitoring By Lc-Msn-It-Tof, Tyler C. Melton, Stacy D. Brown

ETSU Faculty Works

Pharmaceutical compounds represent a rapidly emerging class of environmental contaminants. Such compounds were recently classified by the U.S. Geological Survey, including several antibiotics. An LC-MS/MS screening method for the top five antibiotics in drinking water was developed and validated using a Shimadzu LC-MS-IT-TOF. The separation was performed using a Waters Acquity UPLC BEH C18 column with a gradient elution. Sulfamethazine was exposed to conditions intended to mimic drinking water chlorination, and samples were collected and quenched with excess sodium sulfite. Kinetics of sulfamethazine degradation was followed as well as the formation of the major chlorinated byproduct (m/z 313). For the …

Quantification Of Lansoprazole In Oral Suspension By Ultra-High-Performance Liquid Chromatography Hybrid Ion-Trap Time-Of-Flight Mass Spectrometry, Stacy D. Brown, Justin D. Connor, Nicholas C. Smallwood, Ralph A. Lugo

ETSU Faculty Works

An LC-MS/MS method was developed and validated to be used as a stability indicating assay for the study of a 3 mg/mL lansoprazole oral suspension. The method utilizes a UPLC (ultra-performance liquid chromatography) column and unique mass spectrometric detection (ion-trap time-of-flight (IT-TOF)) to achieve a sensitive (LOD 2 ng/mL), accurate, and reproducible quantification of lansoprazole. This method reports an intraday and interday coefficient of variation of 2.98 ± 2.17% ( for each concentration for each day) and 3.07 ± 0.89% ( for each concentration), respectively. Calibration curves (5–25 μg/mL) were found to be linear with an value ranging from …

Predicting Flavonoid Ugt Regioselectivity With Graphical Residue Models And Machine Learning., Arthur Rhydon Jackson

Electronic Theses and Dissertations

Machine learning is applied to a challenging and biologically significant protein classification problem: the prediction of flavonoid UGT acceptor regioselectivity from primary protein sequence. Novel indices characterizing graphical models of protein residues are introduced. The indices are compared with existing amino acid indices and found to cluster residues appropriately. A variety of models employing the indices are then investigated by examining their performance when analyzed using nearest neighbor, support vector machine, and Bayesian neural network classifiers. Improvements over nearest neighbor classifications relying on standard alignment similarity scores are reported.

Characterization Of The Genes Involved In Biosynthesis And Transport Of Schizokinen, A Siderophore Produced By Rhizobium Leguminosarum Iari 917., David Jack Hammond

Electronic Theses and Dissertations

Iron is the 4th most abundant metal on the earth's crust and is required by most organisms as a cofactor for many enzymes; however, at physiological pH and aerobic conditions iron forms insoluble ferric oxyhydroxide polymers. Siderophores are low molecular weight compounds that scavenge ferric ions, bind with high affinity, and transport it into the cell via multicomponent transport systems. Rhizobia are soil dwelling organisms that form symbiotic relationships with host plants and fix atmospheric nitrogen, while the bacteria receive nutrients. R. leguminosarum IARI 917 produces a siderophore characterized as 'schizokinen'. In the present study, we have characterized the binding …

Sex Differences In Nicotine-Conditioned Hyperactivity In A Model Of Dopamine D2 Receptor Priming: Roles Of Dopamine D2 And D3 Receptor Subtypes., Ashley Brianna Sheppard

Electronic Theses and Dissertations

The aim of this investigation was to determine the effect of a nicotine-conditioned context on locomotor hyperactivity in an animal model of D2-priming, and whether conditioned hyperactivity could be blocked by the D2 antagonist eticlopride or the D3 antagonist nafadotride. D2-primed male rats showed enhanced nicotine sensitization as evidenced by statistically significant differences in horizontal activity. D2-primed female rats administered nicotine demonstrated an increased hypoactive response after initial sensitization and increased stereotypy. Eticlopride and nafadotride blocked sensitization to nicotine in both D2-primed and non D2-primed males and females. Eticlopride blocked conditioned hyperactivity in females but not in males. D2-primed female …

Toward The Synthesis Of Nuclease Models., Enni Nina Fomumbod

Electronic Theses and Dissertations

Nucleases are enzymes that can specifically recognize nucleic acids and hydrolyze their phosphodiester bonds effectively. As is the case with many hydrolases, nucleases often carry one or more metal centers. Cooperation between such metal centers and other interactions involving general acid-base activities are believed to be essential in multifunctional catalyses. Combination of such interactions in model compounds often resulted in larger than additive effects.

This work is aimed at synthesizing nuclease models that combine the ability to recognize phosphate groups and/or nitrogen bases of DNA together with the ability to catalyze phosphodiester hydrolysis. These models were designed to achieve optimum …

Expression, Purification, And Characterization Of The Mast Cell Proteases Chymase And Cathepsin G., Brent E. Lockhart

Electronic Theses and Dissertations

Human mast cells have been associated with wound healing, allergies, inflammation, and defense against pathogens and have been detected in tissues close to blood vessels especially in the areas between the inside of the body and the external environment, such as the skin, lungs, digestive tract, mouth, and nose. Previous studies have shown that mast cells contain large granules filled with histamine, heparin, cytokines, eicosanoids, and the serine proteases, tryptase, Chymase, and cathepsin G (CatG). These proteases are stored and released from mast-cell granules upon activation by antigen binding to IgE immunoglobulins on the cell surface or by direct injury. …

Antibiotics That Inhibit 30s Or 50s Ribosomal Subunit Formation: Hygromycin B, Quinupristin-Dalfopristin And Xrp 2868., Susan Mabe Mcgaha

Electronic Theses and Dissertations

Several antibiotics that prevent translation by binding to ribosomal subunits have been shown to also inhibit ribosomal subunit assembly (Champney and Tober 2003). The aminoglycoside hygromycin B was examined in Escherichia coli cells for inhibitory effects on translation and ribosomal subunit assembly. The streptogramin antibiotics quinupristin-dalfopristin and XRP 2868 (NXL 103) were examined for similar effects on these 2 cellular functions in antibiotic-resistant strains of Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae.

Pulse chase experiments were performed which verified slower rates of ribosomal subunit formation in drug treated cells. Hygromycin B exhibited a concentration dependent inhibitory effect …

Localization Of A Microsporidia Adam (A Disintegrin And Metalloprotease Domain) Protein And Identification Of Potential Binding Partners., Carrie E. Jolly

Electronic Theses and Dissertations

Microsporidia are spore-forming, obligate intracellular pathogens typically associated with opportunistic infections in immunocompromised individuals. Treatment options for microsporidia infections in humans are limited and additional research is necessary to create better therapeutic agents. For many pathogenic organisms, adhesion to the host cell surface is a prerequisite for tissue colonization and invasion. Our previous research has demonstrated a direct relationship between adherence of microsporidia spores to the surface of host cells and infectivity in vitro. In an effort to better understand adherence, we have turned our attention to determining what proteins may be involved in this process. Examination of the …

The Effects Of Nicotine Conditioned Place Preference In D2 Primed Adolescent Rats: Age-Related And Gender Effects., Yoko Emily Ogawa

Electronic Theses and Dissertations

This study investigated nicotine conditioned place preference (CPP) in two different ages of adolescence using a rodent model of schizophrenia. Both 2- and 3-chambered CPP apparatuses were used to test whether the CPP was due to an aversion to the white chamber. Animals were neontally treated with the dopamine D₂/D₃ agonist, quinpirole, or saline and raised to either early postweanling age (P 22) or adolescence (P 29). Rats were conditioned to prefer the white chamber using nicotine. Results showed that nicotine induced CPP and appeared to alleviate an increased stress response in D₂ primed animals, which …

An Analysis Of Nicotine Exacerbation Of Reductions In Ppi In A Rodent Model Of Schizophrenia., Amanda Marie Maple

Electronic Theses and Dissertations

Prepulse inhibition (PPI) is an operational measure of sensorimotor gating and is known to be reduced when the dopamine D² receptor is activated. We used a rodent model of psychosis in which increases in dopamine D_{2 receptor sensitivity are produced through neonatal quinpirole (a dopamine D2 / D3 agonist) treatment to rats. Rats were administered quinpirole (1mg/kg) or saline from postnatal day (P) 1-21. Rats were raised to adulthood and tested on PPI. Results showed that neonatal quinpirole treatment produced a significant reduction in PPI, and nicotine exacerbated this reduction. This reduction was partially blocked by …}

Nicotine Sensitization In A Rodent Model Of Schizophrenia: A Comparison Of Adolescents, Adults, And Neurotrophic Factors., Marla Kay Perna

Electronic Theses and Dissertations

The behavioral effects of nicotine on locomotor activity in a rodent model of psychosis were analyzed. This model is based on neonatal quinpriole treatment (a dopamine D2/D3 agonist) which causes increased D2 receptor sensitivity, a phenomenon known as D2 priming that is common in schizophrenia. D2-primed adolescent rats did not demonstrate nicotine-induced hypoactivity early in training, and males demonstrated more rapid sensitization to nicotine as compared to controls administered nicotine. D2-primed females administered nicotine demonstrated increased stereotypic behavior. D2-primed adult rats given nicotine demonstrated significantly more robust sensitization to nicotine than controls given nicotine. Brain-derived neurotrophic factor (BDNF) was analyzed …