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Discovery And Characterization Of Cytomegalovirus Inhibitors Using Reporter-Based Antiviral Assays, Amine Ourahmane

Theses and Dissertations

ABSTRACT

DISCOVERY AND CHARACTERIZATION OF CYTOMEGALOVIRUS INHIBITORS

USING REPORTER-BASED ANTIVIRAL ASSAYS

By Amine Ourahmane, MS

A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science at Virginia Commonwealth University.

Virginia Commonwealth University, August 2017

Major Director: Michael McVoy, Ph.D.

Professor, Department of Pediatrics and Microbiology and Immunology

Human cytomegalovirus (HCMV), a member of the herpesvirus family, causes significant disease in immunocompromised patients and is the major infectious cause of birth defects when acquired congenitally. Current HCMV antivirals are suboptimal due to modest potency, significant toxicities, and emergence of resistance. Because HCMV does not infect …

Depressive Symptoms Among College Students: An Exploration Of Fundamental Cause Theory, Peter A. Carrico

Theses and Dissertations

Fundamental Cause Theory suggests that socioeconomic and demographic factors are causal to various illnesses, including depression. However, no known previously existing research has used Fundamental Cause Theory to create a model of depression among college students. To do this, the present study conducted a stepwise binomial logistic regression to examine how socioeconomic status and the sociodemographic variables of Gender, Race, and Sexual Orientation, and others predict depressive symptoms in a large sample of undergraduates when controlling for stressful life events and social support (N = 2,915). Results support the hypothesis that socioeconomic disparities in depressive symptoms are the result …

A Systems Biology Approach For Predicting Essential Genes And Deciphering Their Dynamics Under Stress In Streptococcus Sanguinis, Fadi El-Rami

Theses and Dissertations

Infectious diseases are the top leading cause of death worldwide. Identifying essential genes, genes indispensable for survival, has been proven indispensable in defining new therapeutic targets against pathogens, major elements of the minimal set genome to be harnessed in synthetic biology, and determinants of evolutionary relationships of phylogenetically distant species. Thus, essentiality studies promise valuable revenues that can decipher much of biological complexities.

Taking advantage of the available microbial sequences and the essentiality studies conducted in various microbial models, we proposed a framework for the prediction of essential genes based on our experimentally verified knowledge of the pathways involved in …

Health Aid In Africa: Placement, Service Utilization, And Benefit, Carrie Dolan

Theses and Dissertations

While the health sector has attracted significant foreign aid, evidence on the effectiveness of this support is mixed. This dissertation examines the allocation of health aid within the context of placement, service utilization, and benefit. The first paper examined the sub-national allocation of Chinese development aid projects across Africa. I determined how political preferencing of Chinese aid specifically, allocating aid to the birth region of the current political leader differs across sectors such as health, education, and transportation. I find some evidence that aid, more broadly defined, is subject to political preferencing in recipient countries, which could potentially limit its …

Characterization Of A Putative Hemolysin Expressed By Sneathia Amnii, A Preterm Birth-Associated Pathogen, Lizette Carrasco, Kimberly Jefferson

UROP Posters

The gram-negative bacteria Sneathia amnii is a poorly-characterized commensal of the female urogenital tract frequently associated with adverse clinical outcomes such as bacterial vaginosis (BV), amnionitis, and preterm labor. To investigate its potential role in virulence, we sought to identify and characterize virulence determinants produced by S. amnii in an effort to better understand the pathogenesis of infectious preterm birth. Through sequencing of the Sn35 genome (type strain of S. amnii), we identified two genes with amino acid sequence similarity and structural similarity to the filamentous hemagglutinin (FHA) protein of Bordetella pertussis and its Type Vb transporter. Because S. amnii …

Mast Cell Activation By Diverse Stimuli Can Be Suppressed By Steroid Therapy And Targeting The Fyn-Stat5b Cascade, Anuya Paranjape

Theses and Dissertations

Mast cells are critical effectors of allergic disease that can be activated by numerous stimuli. We have examined mast cell control by the inflammatory cytokine, IL-33, as well as IgG. In the first study reported here, we found that the synthetic glucocorticoid, dexamethasone, potently and rapidly suppressed IL-33-induced cytokine production from murine bone marrow–derived and peritoneal mast cells, as well as human mast cells. Dexamethasone also antagonized IL-33-mediated enhancement of IgE-induced cytokine production and migration. Although dexamethasone had no effect on IL-33-induced phosphorylation of MAP kinases or NFκB p65 subunit, it antagonized AP-1 and NFκB-mediated transcriptional activity. Finally, intraperitoneal administration …

Comparing The Structural Components Variance Estimator And U-Statistics Variance Estimator When Assessing The Difference Between Correlated Aucs With Finite Samples, Anna L. Bosse

Theses and Dissertations

Introduction: The structural components variance estimator proposed by DeLong et al. (1988) is a popular approach used when comparing two correlated AUCs. However, this variance estimator is biased and could be problematic with small sample sizes.

Methods: A U-statistics based variance estimator approach is presented and compared with the structural components variance estimator through a large-scale simulation study under different finite-sample size configurations.

Results: The U-statistics variance estimator was unbiased for the true variance of the difference between correlated AUCs regardless of the sample size and had lower RMSE than the structural components variance estimator, providing better type 1 error …

Role Of Mir-155 And Mir-146a In Mast Cell Function, Amina Abdul Qayum

Theses and Dissertations

Mast cells are resident immune cells abundantly found in the tissue at the host-environment interface, where they play a critical role in inflammatory allergic responses. Mast cell responses may be regulated by the cytokine milieu at the site of inflammation. Recent studies have revealed microRNAs to be important in altering cytokine signaling in immune cells. Here, we demonstrate for the first time that IL-10 and IL-33 induce miR-155 and miR-146a, respectively, to alter mast cell functions. We report that IL-10 enhanced IgE induced activation of mast cells. IL-10 effects are dependent on Stat3 activation, which elicits miR-155 expression, resulting in …

Role Of The Pediatric Dental Provider In Human Papillomavirus (Hpv) Education, Meredith L. Dugoni

Theses and Dissertations

Purpose: This study investigates knowledge about HPV and examines if pediatric dental providers should include HPV education for guardians of patients 10-18 years.

Methods: Legal guardians of 10-18 year-old patients of the Virginia Commonwealth University Pediatric Dental Clinic were enrolled in this prospective cohort study. Participants completed a baseline survey, were provided HPV education, completed an initial follow-up survey, and then completed a 6-month follow-up survey.

Results: A total of 54 participants completed the baseline and initial follow-up surveys and 17 completed the 6-month follow-up survey. The average number of correct responses was 3.4 of 6 knowledge questions, which significantly …

The Role Of The Nucleosome Remodeling Factor Nurf In Inhibiting T And Natural Killer Cell Mediated Antitumor Immunity By Suppressing Tumor Antigenicity And Natural Cytotoxicity Receptor Co-Ligands, Kimberly Mayes

Theses and Dissertations

Tumor immunoediting is a dynamic process in which the immune response attacks tumor cells by detecting danger signals and tumor antigens. In order to survive, tumor cells develop mechanisms to avoid detection or destruction by the immune system. To counteract this, several strategies are being developed to enhance the antitumor immune response, including the depletion of immunosuppressive cells, enhancing the activation of antitumor immune cells and increasing tumor cell immunogenicity. These therapies have seen limited success individually, however, and it is likely that combination therapy with novel targets will be necessary to see reproducible beneficial responses. Epigenetic modifications are attractive …

Functional Study Of Cmya5, A Candidate Gene For Schizophrenia, Anting Hsiung

Theses and Dissertations

CMYA5 is a candidate gene for schizophrenia because of the association of variant rs10043986 (Pro4063Leu). Studies of CMYA5 and its gene product, myospryn, in brain and neuronal cells have not been previously reported. We examined the neuronal expression of myospryn and its binding partner, desmin intermediate filament (IF), and investigated the difference in binding and colocalization of the two alleles of myospryn to IFs. Myospryn and desmin are expressed in brain regions. Using yeast two-hybrid and surface plasmon resonance, the T allele (Leu) is found to have higher binding affinity to desmin than the C allele (Pro). Myospryn localizes to …

A Targeted Approach To Increasing The African American Blood Donor Pool, Arnethea L. Sutton

Theses and Dissertations

A continuous need for blood products, specifically for those who require frequent transfusions, such as individuals with sickle cell disease, warrants the need for targeted interventions to increase blood donations from underrepresented populations. One population in particular, African Americans, only account for 1% of blood donors in the United States. Literature indicates numerous reasons why this population is underrepresented amongst donors, including fear, lack of knowledge about the blood donation, and specific to this population, lack of trust in the medical community. This study involves the development, implementation, and assessment of a targeted educational approach, incorporating the Theory of Planned …

A Multi-Level Model Of Synergistic T Cell Activation, Anna Konstorum, Anthony Vella, Reinhard C. Laubenbacher

Biology and Medicine Through Mathematics Conference

No abstract provided.

Anticipating Elimination Of Mosquito-Borne Diseases, Suzanne M. O'Regan, Jonathan Lillie, John M. Drake

Biology and Medicine Through Mathematics Conference

No abstract provided.

Toward Adaptive Control Of Acute Inflammation, Judy D. Day, Seddik M. Djouadi, Ouassim Bara, Gregory L. Zitelli

Biology and Medicine Through Mathematics Conference

No abstract provided.

Mathematical Modeling Of Endotoxin-Induced Inflammation In Young Men, Renee Brady

Biology and Medicine Through Mathematics Conference

No abstract provided.

Mathematical Models Of Hiv And Hpv Coinfection, Samantha Erwin, Meghna Verma, Vida Abedi, Raquel Hontecillas-Magarzo, Stefan Hoops, Josep Bassaganya-Riera, Stanca M. Ciupe

Biology and Medicine Through Mathematics Conference

No abstract provided.

Can Including Time Delay In Epidemic Models Significantly Improve Predictions Concerning Intervention Strategies?, Adrienna N. Bingham, Leah Shaw

Biology and Medicine Through Mathematics Conference

No abstract provided.

Needle In The Haystack: Combining Intravital Imaging And Mathematical Modeling To Understand How Vaccine-Induced T Cells Find Malaria-Infected Cells In Murine Livers, Vitaly V. Ganusov, Ian Cockburn, Reka Kelemen

Biology and Medicine Through Mathematics Conference

No abstract provided.

General Models For Ecological Drivers Of Poverty, Calistus Ngeh Ngonghala

Biology and Medicine Through Mathematics Conference

No abstract provided.

Cryptosporidium Priming Is More Effective Than Vaccine For Protection Against Cryptosporidiosis In A Murine Protein Malnutrition Model, Luther A. Bartelt, David T. Bolick, Glynis L. Kolling, James K. Roche, Edna I. Zaenker, Ana M. Lara, Francisco Jose Noronha, Carrie A. Cowardin, John H. Moore, Jerrold R. Turner, Cirle A. Warren, Gregory A. Buck, Richard L. Guerrant

Biochemistry and Molecular Biology Publications

Cryptosporidium is a major cause of severe diarrhea, especially in malnourished children. Using a murine model of C. parvum oocyst challenge that recapitulates clinical features of severe cryptosporidiosis during malnutrition, we interrogated the effect of protein malnutrition (PM) on primary and secondary responses to C. parvum challenge, and tested the differential ability of mucosal priming strategies to overcome the PM-induced susceptibility. We determined that while PM fundamentally alters systemic and mucosal primary immune responses to Cryptosporidium, priming with C. parvum (106 oocysts) provides robust protective immunity against re-challenge despite ongoing PM. C. parvum priming restores mucosal Th1-type effectors (CD3+CD8+CD103+ T-cells) …

Regulation Of Reactive Oxygen Species And The Antioxidant Protein Dj-1 In Mastocytosis, Do-Kyun Kim, Michael A. Beaven, Joseph M. Kulinski, Avanti Desai, Geethani Bandara, Yun Bai, Calman Prussin, Lawrence B. Schwartz, Hirsh Komarow, Dean D. Metcalfe, Ana Olivera

Internal Medicine Publications

Neoplastic accumulation of mast cells in systemic mastocytosis (SM) associates with activating mutations in the receptor tyrosine kinase KIT. Constitutive activation of tyrosine kinase oncogenes has been linked to imbalances in oxidant/antioxidant mechanisms in other myeloproliferative disorders. However, the impact of KIT mutations on the redox status in SM and the potential therapeutic implications are not well understood. Here, we examined the regulation of reactive oxygen species (ROS) and of the antioxidant protein DJ-1 (PARK-7), which increases with cancer progression and acts to lessen oxidative damage to malignant cells, in relationship with SM severity. ROS levels were increased in both …

The Pathogen-Occupied Vacuoles Of Anaplasma Phagocytophilum And Anaplasma Marginale Interact With The Endoplasmic Reticulum, Hilary K. Truchan, Chelsea L. Cockburn, Kathryn S. Hebert, Forgivemore Magunda, Susan M. Noh, Jason A. Carlyon

Microbiology and Immunology Publications

The genus Anaplasma consists of tick-transmitted obligate intracellular bacteria that invade white or red blood cells to cause debilitating and potentially fatal infections. A. phagocytophilum, a human and veterinary pathogen, infects neutrophils to cause granulocytic anaplasmosis. A. marginale invades bovine erythrocytes. Evidence suggests that both species may also infect endothelial cells in vivo. In mammalian and arthropod host cells, A. phagocytophilum and A. marginale reside in host cell derived pathogen-occupied vacuoles (POVs). While it was recently demonstrated that the A. phagocytophilum-occupied vacuole (ApV) intercepts membrane traffic from the trans-Golgi network, it is unclear if it or …

Involvement Of Nadh Oxidase In Biofilm Formation In Streptococcus Sanguinis, Xiuchun Ge, Xiaoli Shi, Limei Shi, Jinlin Liu, Victoria Stone, Fanxiang Kong, Todd Kitten, Ping Xu

Microbiology and Immunology Publications

Biofilms play important roles in microbial communities and are related to infectious diseases. Here, we report direct evidence that a bacterial nox gene encoding NADH oxidase is involved in biofilm formation. A dramatic reduction in biofilm formation was observed in a Streptococcus sanguinis nox mutant under anaerobic conditions without any decrease in growth. The membrane fluidity of the mutant bacterial cells was found to be decreased and the fatty acid composition altered, with increased palmitic acid and decreased stearic acid and vaccenic acid. Extracellular DNA of the mutant was reduced in abundance and bacterial competence was suppressed. Gene expression analysis …

Interaction Between Cannabinoid System And Toll-Like Receptors Controls Inflammation, Kathleen L. Mccoy

Microbiology and Immunology Publications

Since the discovery of the endocannabinoid system consisting of cannabinoid receptors, endogenous ligands, and biosynthetic and metabolizing enzymes, interest has been renewed in investigating the promise of cannabinoids as therapeutic agents. Abundant evidence indicates that cannabinoids modulate immune responses. An inflammatory response is triggered when innate immune cells receive a danger signal provided by pathogen- or damage-associated molecular patterns engaging pattern-recognition receptors. Toll-like receptor family members are prominent pattern-recognition receptors expressed on innate immune cells. Cannabinoids suppress Toll-like receptor-mediated inflammatory responses. However, the relationship between the endocannabinoid system and innate immune system may not be one-sided. Innate immune cells express …

Interaction Between Cannabinoid System And Toll-Like Receptors Controls Inflammation, Kathleen L. Mccoy

Microbiology and Immunology Publications

Since the discovery of the endocannabinoid system consisting of cannabinoid receptors, endogenous ligands, and biosynthetic and metabolizing enzymes, interest has been renewed in investigating the promise of cannabinoids as therapeutic agents. Abundant evidence indicates that cannabinoids modulate immune responses. An inflammatory response is triggered when innate immune cells receive a danger signal provided by pathogen- or damage-associated molecular patterns engaging pattern-recognition receptors. Toll-like receptor family members are prominent pattern-recognition receptors expressed on innate immune cells. Cannabinoids suppress Toll-like receptor-mediated inflammatory responses. However, the relationship between the endocannabinoid system and innate immune system may not be one-sided. Innate immune cells express …

Mir-9 Acts As An Oncomir In Prostate Cancer Through Multiple Pathways That Drive Tumour Progression And Metastasis, S. J. Seashols-Williams, W. Budd, G. C. Clark, Q. Wu, R. Daniel, E. Dragoescu, Z. E. Zehner

Biochemistry and Molecular Biology Publications

Identification of dysregulated microRNAs (miRNAs) in prostate cancer is critical not only for diagnosis, but also differentiation between the aggressive and indolent forms of the disease. miR-9 was identified as an oncomiR through both miRNA panel RT-qPCR as well as high-throughput sequencing analysis of the human P69 prostate cell line as compared to its highly tumorigenic and metastatic subline M12, and found to be consistently upregulated in other prostate cell lines including DU-145 and PC3. While miR-9 has been characterized as dysregulated either as an oncomiR or tumour suppressor in a variety of other cancers including breast, ovarian, and nasopharyngeal …

Impaired Immunological Synapse In Sperm Associated Antigen 6 (Spag6) Deficient Mice, Lauren Folgosa Cooley, Mohey Eldin El Shikh, Wei Li, Rebecca C. Keim, Zhengang Zhang, Jerome F. Strauss, Zhibing Zhang, Daniel H. Conrad

Microbiology and Immunology Publications

Sperm associated antigen 6 (SPAG6), a component of the central apparatus of the “9 + 2” axoneme, plays a central role in ciliary and flagellar motility; but, its contribution to adaptive immunity and immune system development is completely unknown. While immune cells lack a cilium, the immunological synapse is a surrogate cilium as it utilizes the same machinery as ciliogenesis including the nucleation of microtubules at the centrosome. This prompted our hypothesis that SPAG6 critically regulates the formation and function of immunological synapses. Using bone marrow reconstitution studies of adult WT mice, we demonstrate that SPAG6 is expressed in primary …

The Prostaglandin E2-Ep3 Receptor Axis Regulates Anaplasma Phagocytophilum-Mediated Nlrc4 Inflammasome Activation, Xiaowei Wang, Dana K. Shaw, Holly L. Hammond, Fayyaz S. Sutterwala, Manira Rayamajhi, Kari Ann Shirey, Darren J. Perkins, Joseph V. Bonventre, Thangam S. Velayutham, Sean M. Evans, Kyle G. Rodino, Lauren Viebrock, Karen M. Scanlon, Nicholas H. Carbonetti, Jason A. Carlyon, Edward A. Miao, Jere W. Mcbride, Michail Kotsyfakis, Joao H. F. Pedra

Microbiology and Immunology Publications

Rickettsial agents are sensed by pattern recognition receptors but lack pathogen-associated molecular patterns commonly observed in facultative intracellular bacteria. Due to these molecular features, the order Rickettsiales can be used to uncover broader principles of bacterial immunity. Here, we used the bacterium Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis, to reveal a novel microbial surveillance system. Mechanistically, we discovered that upon A. phagocytophilum infection, cytosolic phospholipase A2 cleaves arachidonic acid from phospholipids, which is converted to the eicosanoid prostaglandin E2 (PGE2) via cyclooxygenase 2 (COX2) and the membrane associated prostaglandin E synthase-1 (mPGES-1). PGE2-EP3 receptor signaling leads …

Aftermath Of The Hobby Lobby Decision: Implications For Women In The Workforce, Hirsh Shah

AUCTUS: The Journal of Undergraduate Research and Creative Scholarship

Hobby Lobby is a chain of 640 arts and crafts stores owned by the Green family, based in Oklahoma City. This company is required to follow the Affordable Care Act (ACA), which mandates that larger employers—those with more than 50 employees—have to include coverage for the full range of preventative care, including contraceptives, in their female employees’ health insurance plans. However, the Green family holds deeply religious views and did not want to include four of the twenty contraceptives covered by the ACA, including long acting reversible contraception and emergency contraception, in their female employee coverage. The family believed that …