Life Sciences Commons^™

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 …

Exploring The Structure And Biochemistry Of Oxidation-Mediated Inhibitation Of The Peptidyl-Prolyl Isomerase Pin1, Brendan T. Innes

Electronic Thesis and Dissertation Repository

Pin1 is a phosphorylation-dependent peptidyl-prolyl isomerase that has been shown to be neuroprotective in aging-related neurodegenerative diseases such as Alzheimer's disease (AD). However, it is not active in AD brain, and a recent proteomic screen of Mild Cognitive Impairment (MCI) brain samples revealed that Pin1 is oxidized in the brains of these pre-AD patients. This suggests that this oxidation may be the cause of the loss of the neuroprotective Pin1 function in AD. The Pin1 active site contains a functionally critical cysteine residue (Cys113) with a low predicted pKa, making it highly susceptible to oxidation. We hypothesize that Pin1 is …

Prenatal Programming Of Hepatic Glucose And Cholesterol Regulation In Male Rat Offspring By Chronic Intermittent Hypoxia, Waseem Iqbal

Electronic Thesis and Dissertation Repository

Obstructive sleep apnea (OSA) is a chronic disorder involving repetitive interruptions in breathing during sleep. Sufferers of OSA are exposed to chronic intermittent hypoxia (CIH), characterized by cyclical reductions in oxygen availability. A number of studies have established a link between OSA and various cardiovascular and metabolic comorbidities in adulthood, including hypertension, obesity, and type II diabetes. While the consequences of OSA in adults have been well described, the cross-generational impact of this condition and potential effects on fetal development are not known. Epidemiological and animal studies have demonstrated that physiological insults during pregnancy lead to diminished growth of offspring …

The Staphylococcus Pseudintermedius Adhesin Spsd Contains A Central Fibronectin-Binding Domain, Andrea S. Bordt

Dissertations & Theses (Open Access)

Staphylococcus pseudintermedius is a Gram-positive bacterium significant because of its ability to cause costly and difficult to treat veterinary infections worldwide. It exhibits several similarities to Staphylococcus aureus, however, very little is known about its surface adhesins. Surface adhesins in S. aureus are significant contributors to pathogenesis. S. pseudintermedius encodes the surface protein SpsD, which contains characteristics of the microbial surface components recognizing adhesive matrix molecules family and confers attachment of the heterologous host Lactococcus lactis to fibronectin. This work has identified a centrally-located fibronectin binding domain in SpsD which binds the 30 kDa N-terminal domain of fibronectin with …

In Vitro Analysis Of Short Chain Fatty Acids And Human Fecal Microbiota Stimulated By Pectin Sources, Byungjick Min

Graduate Theses and Dissertations

As interest increases for colonic health, various studies have been conducted to investigate functional ingredients for host health and prevention of chronic disease in humans such as colon cancer and Type 2 diabetes. In particular dietary fiber has been reported to have beneficial effects on colonic health. More recently, dietary fiber has been categorized based on its characteristics and numerous studies have been conducted to determine correlations with chronic diseases. Pectin is a naturally occurring biopolymer that has been used in various applications for purposes of pharmaceutical and biotechnology. Pectin is a soluble dietary fiber and can be entirely fermented …

Phenotypic, Physiological And Growth Interactions Among Salmonella Serovars, Juliany Rivera Calo

Graduate Theses and Dissertations

This thesis consists of four research parts: a literature review that covers Salmonella spp., one of the more prominent foodborne pathogens that represents a major risk to humans (chapter 1). Understanding the growth of Salmonella serovars and strains is an important basis for more in depth research. In this case we studied a) the aerobic and anaerobic growth responses of multiple strains from six different serovars, b) how the spent media from different serovars, more importantly S. Heidelberg, affect the growth of S. Typhimurium, and c) determined whether or not two different serovars undergo competitive interactions when they were …

Effect Of Electrostatic Spraying Of Food-Grade Organic And Inorganic Acids, And Grape Seed Extract On Salmonella Typhimurium And Escherichia Coli O157:H7 On Spinach And Iceberg Lettuce, Vijayalakshmi Ganesh

Graduate Theses and Dissertations

The safety of fresh produce is often challenged by the sporadic incidences of outbreaks owing to Salmonella Typhimurium (S.T.) and Escherichia coli O157:H7 (E.c.). Washing and current disinfection strategies do not completely decontaminate leafy greens such as spinach and iceberg lettuce. Therefore, more effective treatments are essential to ensure the microbial safety of the produce at the selling point. Here, a multiple hurdle approach was applied: (1) Natural antimicrobials - organic acids (malic, lactic and tartaric acids; MA, LA, TA, respectively) and plant extract (grape seed extract, GSE) and (2) Application method - Electrostatic spraying, was …

Electrostatic Spray Application Of Food-Grade Organic Acids And Plant Extracts To Decrease Escherichia Coli O157:H7 And Salmonella Typhimurium On Select Produce, Leighanna Maeree Massey

Graduate Theses and Dissertations

Both consumers and suppliers have been negatively affected by an increase in foodborne pathogens contaminating fruits. Consequently, there is a need for the development of more efficient antimicrobials and application techniques to decrease contamination. Natural preservatives such as organic acids and plant extracts have demonstrated promising results in decontaminating produce. In addition, the effectiveness of such preservatives may be enhanced by the use of an electrostatic sprayer. The objective of this research was to determine the combinations and concentrations of organic acids and plant extract that were able to reduce Salmonella Typhimurium (ST) and Escherichia coli O157:H7 (EC) inoculated fruits …

Identification Of Biomarkers Associated With Ascites Incidence In Broilers, Kaylee Rowland

Graduate Theses and Dissertations

Poultry is key in genetic research due to breeding feasibility, relatively short generation interval, and distinct phenotypes. It is estimated that 8% of broiler deaths annually can be attributed to ascites, an economically important disease that has been challenging the industry for the past 2 decades. Genetically selected ascites resistant (RES) and susceptible (SUS) chicken lines have been established and maintained by the University of Arkansas, Fayetteville. Intensive research efforts have been made to reveal physiological and biochemical characteristics for the incidence of ascites. Since the whole genome of the major ancestral chicken, the Red Jungle Fowl, has been sequenced, …

Imaging Breast Cancer Progression And Lymph Node Metastases In Murine Models Using Mri And Magnetic Nanoparticles, Vasiliki Economopoulos

Electronic Thesis and Dissertation Repository

Most breast cancer related deaths are caused by the spread or metastasis of the primary tumor to distant sites in the body. The lymph nodes are one of the first places where metastases can be detected and are frequently examined for macroscopic metastases to help determine course of treatment for patients. However, little is known about the significance of microscopic metastases and disseminated individual cancer cells within the nodes. The goal of this work was to use MRI to monitor the development of primary tumors and lymphatic metastases in models of breast cancer.

In this thesis, we examined the MRI …

Regulating Rsma Expression In Pseudomonas Aeruginosa, Sean D. Stacey

Electronic Theses and Dissertations

Pseudomonas aeruginosa, a Gram-negative bacillus, commonly infects immunocompromised individuals and uses a variety of virulence factors to persist in these hosts. The posttranscriptional regulator, RsmA, plays a role in the expression of many virulence factors in P. aeruginosa. RsmA up regulates virulence factors used in colonizing hosts. However, regulation of rsmA is not well elucidated. Transposon mutagenesis was performed on P. aeruginosa containing a transcriptional rsmA-lacZ fusion to answer this question. Mutants were screened via β-galactosidase assay and transposon insertions identified via arbitrary PCR. A probable MFS transporter, we named mtpX, was one significant transposon mutant identified. …

Exercise Participation During Weight Loss On A High Protein – Low Carbohydrate Diet Plan In Females Aged 15-25 Years, Margaret Mobley-Meulman

Electronic Theses and Dissertations

Weight gain due to poor diet and lack of exercise is responsible for over 300,000 deaths each year (U.S. Department of Health and Human Services, 2010). Obese adults have an increased risk for serious health conditions including high blood pressure and cholesterol, type 2 diabetes, coronary heart disease, stroke, gallbladder disease, osteoarthritis, sleep apnea, respiratory problems, and certain cancers (National Cancer Institute, 2012). Participation in exercise can help control weight, strengthen muscles and bones, and reduce the incidence of cardiac events, stroke, hypertension, type 2 diabetes, colon and breast cancers, osteoporotic fractures, gallbladder disease, obesity, depression, anxiety, and delay mortality …

Comparison Of Potential Salmonella Portals Of Entry And Tissue Distribution Following Challenge Of Poultry, Gopala Krishna Kallapura Sheshagiri

Graduate Theses and Dissertations

The following studies evaluated our hypothesis that transmission by the fecal-respiratory route may be a viable portal of entry for Salmonella and could explain some clinical impressions of relatively low-dose infectivity under field conditions in relation to the requisite high oral challenge dose that is typically required for infection of poultry through the oral route in laboratory studies. Initial field reports indicating tracheal sampling to be a sensitive tool for monitoring Salmonella infection in commercial flocks, suggested that tracheal contamination could be a good indicator of Salmonella infection under commercial conditions. Further, a usual assumption regarding airborne Salmonella reaching the …

Relationship Of Global Dna Methylation With Cardiovascular Fitness And Body Composition, Michael Steele Jarrett

UNLV Theses, Dissertations, Professional Papers, and Capstones

Background: Global DNA Methylation (GDM), an epigenomic modification has been linked to the development of Cardiovascular Disease and its risk factors. Purpose: The purpose of this study was to determine if there is a significant correlation between GDM and cardiovascular fitness, as well as, measures of body composition. Methodology: 26 apparently healthy, adults (11 males) completed a physical activity and diet questionnaire, as well as, had a small blood sample (600μL) collected via finger prick for the determination of GDM. Body composition was assessed by means of a Dual-Energy X-ray Absorptiometry (DEXA) scan, while cardiovascular fitness was evaluated by the …

Bioengineering The Expression Of Active Recombinant Human Cathepsin G, Enteropeptidase, Neutrophil Elastase, And C-Reactive Protein In Yeast, Eliot T. Smith

Electronic Theses and Dissertations

The yeasts Pichia pastoris and Kluyveromyces lactis were used to express several recombinant human proteins for further biochemical characterization. Two substitution variants of recombinant human enteropeptidase light chain (rhEPL) were engineered to modify the extended substrate specificity of this serine protease. Both were secreted as active enzymes in excess of 1.7 mg/L in P. pastoris fermentation broth. The substitution variant rhEPL R96Q showed significantly reduced specificities for the preferred substrate sequences DDDDK and DDDDR; however, the rhEPL Y174R variant displayed improved specificities for these substrate sequences relative to all other reported variants of this enzyme. The neutrophil serine proteases human …

T-Cell Treatments For Solid And Hematological Tumors, Drew C. Deniger

Dissertations & Theses (Open Access)

Cell-based therapies have demonstrated potency and efficacy as cancer treatment modalities. T cells can be dichotomized by their T cell receptor (TCR) complexes where alpha/beta T cells (95% of T cells) and gamma/delta T cells (+T cells proliferated to clinically significant numbers and ROR1⁺ tumor cells were effectively targeted and killed by both ROR1-specific CAR⁺ T cell populations, although ROR1RCD137 were superior to ROR1RCD28 in clearance of leukemia xenografts in vivo. The second specific aim focused on generating bi-specific CD19-specific CAR⁺ gamma/delta T cells with polyclonal TCRgamma/delta repertoire on CD19⁺ artificial antigen presenting cells (aAPC). …

Characterization Of Host Immune Responses To Eimeria Adenoeides Infection In Turkey Poults, Ujvala Deepthi Gadde

Graduate Theses and Dissertations

Coccidiosis is a common enteric disease of turkeys that is caused by protozoan parasites belonging to the genus Eimeria. There are about seven species of Eimeria that affect turkeys and of these E. adenoeides is the most pathogenic and commonly recognized. Infection with Eimeria is known to induce a long lasting protective immunity in chickens, but nothing is known regarding the acquisition of immunity to Eimeria in turkeys. The experiments reported here were aimed at investigating the biological and cellular immune response to E. adenoeides in turkey poults under different conditions of exposure. In experiment 1 (Chapter II), 20 …

An Insight Into The Microbial Diversity And Expression Of Cysteine Protease Inhibitors (Cystatin) In Rickettsia Parkeri Infected Amblyomma Maculatum, Khemraj Budachetri

Master's Theses

Amblyomma maculatum (Gulf Coast tick) is an emerging tick species of public health significance in United States. It is a competent vector of Rickettsia parkeri, an etiological agent of a human rickettsiosis. In this study, we investigated the spotted fever group of rickettsial diversity in A. maculatum based on rickettsial ompA gene PCR. Our results showed A. maculatum harbors R. parkeri, R. amblyommii, and R. endosymbiont of A. maculatum. While only R. parkeri was detected in female salivary glands which suggest its ability to traffic from midgut to salivary glands via hemocoel. The presence of R. parkeri was further …

Mechanism Of Lpa-Induced Lipid Uptake In Macrophages, Kan Xu

Masters Theses

Cardiovascular disease, currently the leading cause of mortality throughout the developed countries, is mainly caused by atherosclerosis, which is recognized as a chronic inflammatory disease. Atherosclerotic plaques are characterized by accumulations of lipid in arterial walls together with infiltration of macrophages. These macrophages differentiate from monocytes which transform into foam cells through phagocytizing various forms of lipid, are believed to be the main component of early atherosclerotic lesions.

Lysophosphatidic acid (LPA), a potent bioactive lipid, regulates a broad range of cellular functions in various cell types. In our findings, LPA-induced macrophages may enhance the lipid uptake effect in both J774A.1-mouse …

In Vitro Selection Of Chloroquine Tolerant Plasmodium Falciparum Parasites, Daniel A. Daley

Biological Sciences Theses & Dissertations

Malaria is among the most devastating human diseases, and a majority of lethal cases are caused by the protozoan parasite, Plasmodiumfalciparum. The emergence of multi-drug resistant P.falciparum is a major obstacle to malaria control and is highlighted by the abandonment of chloroquine (CQ) as a first-line treatment of P.falciparum infections worldwide. Chloroquine resistance (CQR) is associated primarily with mutations in the transmembrane digestive vacuole protein, PfCRT. However, CQR P.falciparum parasites harboring the same mutant pfcrt allele vary in their CQ response, suggesting the CQ response is multigenic in nature. No gene outside of pfcrt is completely associated with CQR. Thus, …

Human Adenovirus E1a Binds And Retasks Cellular Hbre1, Blocking Interferon Signalling And Activating Virus Early Gene Transcription, Gregory J. Fonseca

Electronic Thesis and Dissertation Repository

Upon infection, human adenovirus (HAdV) must block interferon signaling and activate the expression of its early genes to reprogram the cellular environment to support virus replication. During the initial phase of infection, these processes are orchestrated by the first HAdV gene expressed during infection, early region 1A (E1A). E1A binds and appropriates components of the cellular transcriptional machinery to modulate cellular gene transcription and activate viral early genes transcription. We have identified hBre1/RNF20 as a novel target of E1A. hBre1 is an E3 ubiquitin ligase which acts with the Ube2b E2 conjugase and accessory factors RNF40 and WAC1 to monoubiquitinate …

Regulation Of Lipid Homeostasis, Inflammatory Signalling And Atherosclerosis By The Peroxisome Proliferator-Activated Receptor Delta, Lazar A. Bojic

Electronic Thesis and Dissertation Repository

The peroxisome proliferator-activated receptor (PPAR) δ is a ligand-dependent transcription factor that has been implicated in metabolic and inflammatory regulation. The molecular and physiological mechanisms by which PPARδ activation regulates lipid metabolism, inflammatory signaling and protection from atherosclerosis in states of metabolic disturbance such as insulin resistance and dyslipidemia, were investigated in a series of in vitro and in vivo studies. In vitro experiments demonstrated that PPARδ activation inhibits atherogenic lipoprotein-induced lipid accumulation and the associated proinflammatory responses. The primary mechanisms for these effects were increased fatty acid β-oxidation, decreased lipoprotein lipase (LPL) activity, reduced MAPK signaling and improved insulin …

Amyloid Beta Resistance And The Warburg Effect: Re-Examining Alzheimer's Disease, Jordan Taylor Newington

Electronic Thesis and Dissertation Repository

Alzheimer’s disease (AD) is characterized by deposition of the amyloid beta (Ab) peptide in the brain, an event which frequently but not universally correlates with nerve cell death. Although most nerve cells die in response to Ab, small populations of cells are able to survive by becoming resistant to Ab toxicity. Understanding the mechanisms by which cells become resistant to Ab may reveal novel treatments for AD. Interestingly, nerve cell lines selected for resistance against Ab exhibit increased glucose uptake and glycolytic flux. Here I show that these metabolic changes are mediated through an upregulation of pyruvate dehydrogenase kinase 1 …

Genetic Approaches To Studying Complex Human Disease, Joseph B. Dube

Electronic Thesis and Dissertation Repository

Common, complex diseases such as cardiovascular disease (CVD) represent an intricate interaction between environmental and genetic factors and now account for the leading causes of mortality in western society. By investigating the genetic component of complex disease etiology, we have gained a better understanding of the biological pathways underlying complex disease and the heterogeneity of complex disease risk. However, the development of high throughput genomic technologies and large well-phenotyped multi-ethnic cohorts has opened the door towards more in-depth and trans-disciplinary approaches to studying the genetics of complex disease pathogenesis. Accordingly, we sought to investigate select complex traits and diseases using …

Use Of A Specific Alpha7 Nicotinic Acetylcholine Receptor Agonist Can Prevent Loss Of Retinal Ganglion Cells In An In Vivo Rat Glaucoma Model, David Mata

Masters Theses

Acetylcholine (ACh) has been shown to have a neuroprotective effect against glutamate-induced excitotoxicity in cultured retinal ganglion cells (RGCs) through activation of α7 nicotinic ACh receptors (nAChRs). In this study, the neuroprotective effects of the α7 nAChR agonist, PNU-282987, were investigated using an in vivo model of glaucoma in adult Long Evans rats. Hypertonic saline injections were used to induce a glaucoma model. After one month, retinas were removed, flat mounted, fixed and labeled with an antibody against Thy 1.1 to label RGCs and fluorescently tagged for visualization. Eye drop application of PNU-282987 resulted in neuroprotection against RGC loss in …

Expression Of The Chemokine Receptor, Cxcr4, And Its Ligand, Sdf-1, Are Increased In Purkinje Cells Of The Multiple System Atrophy Cerebellum, Megan Welter

Masters Theses

Multiple System Atrophy (MSA) is a sporadic, neurodegenerative disease that consists of three conditions: autonomic dysfunction, Parkinsonism and cerebellar ataxia. Our lab conducted an Affymetrix global gene expression analysis using pons tissue of MSA patients to determine genes that are differentially expressed when compared to non- MSA controls. This study identified upregulated genes, including the C-X-C chemokine receptor type 4, CXCR4, to which stromal cell-derived factor-I (SDF-1) is the natural ligand. The CXCR4/SDF-1 signaling pair has been sho.wn to play multiple roles in the brain, such as inducing neuronal apoptosis and promoting leukocyte recruitment during inflammation. The MSA cerebellum presents …

Diabetes Mellitus And Hypercholesterolemia Are Risk Factors For Alzheimer’S Disease And Appear To Affect The Integrity Of The Blood Brain Barrier, Jacqueline Dash

Graduate School of Biomedical Sciences Theses and Dissertations

Studies have shown that the vascular risk factors common to diabetes mellitus and hypercholesterolemia are also risk factors for Alzheimer’s disease (AD). It is currently unknown how these diseases are associated with AD, but they may cause a leak in the blood brain barrier (BBB), which is one of the hallmarks of AD. In this preliminary study, over 150 pig brain slides were tested for the expression levels of tight junction proteins occludin and claudin V in the BBB microvasculature. There were three groups of pig brains used in this study namely, control pigs, pigs with diabetes mellitus and hypercholesterolemia …

Discovery And Characterization Of A Tanapoxvirus Encoded Human Neuregulin Mimetic, David Jeng

Dissertations

Studies on large dsDNA viruses such as poxviruses have been helpful in identifying a number of viral and cellular growth factors that contribute to our broad understanding of virus-host interaction. Orthopoxviruses and leporipoxviruses are among the most studied viruses in this aspect. However, tanapoxvirus (TPV) a member of the genus Yatapoxvirus, still remains largely unexplored, as the only known hosts for this virus are humans and monkeys. Here, we describe the initial characterization of an EGF-like growth factor mimicking human neuregulin from TPV, expressed by the TPV-15L gene. Assays using a baculovirus expressed and tagged TPV-15L protein demonstrated the ability …

Associations Between Alcohol Consumption And Fasting Blood Glucose In Young Adults, Julie Ann Lucca

Master's Theses

Current research shows moderate alcohol consumption is associated with decreased risk of diabetes and excessive consumption or binge drinking can cause insulin resistance and diabetes. In 2010, diabetes was the seventh leading cause of death in the United Statesand was responsible for significant health complications: blindness, kidney failure, and limb amputations, and is a large national economic burden. Fasting blood glucose (FBG) is a tool used to help diagnose diabetes. Abnormally high FBG, ≥100 mg/dl, is indicative of diabetes and pre-diabetes. Few studies have observed diabetic prevalence among young adults or college students. Studying young adults can help provide added …

Oxidation Of Thrombomodulin Methionine 388 In Cigarette Smokers, Samrat B. Thapa

Graduate Theses and Dissertations

This work tested the hypothesis that oxidation of methionine 388 in thrombomodulin is higher in cigarette smokers, and thus a likely contributor towards the hypercoagulable state in smokers. Thrombomodulin, a protein cofactor found on endothelial cell surfaces, regulates the activity of thrombin. Thrombin bound to thrombomodulin no longer converts fibrinogen to fibrin, but instead activates Protein C which, in turn, stops the coagulation cascade by inactivation of clotting factors. The oxidation of methionine 388 of thrombomodulin has been shown in vitro to dramatically decrease the anticoagulant cofactor activity of thrombomodulin. The blood of cigarette smokers is more prone to clot …