Life Sciences Commons^™

Decreasing The Antibiotic Survival Of Multi-Drug Resistant Gram-Positive Organisms, Areej Malik

Biomedical Sciences Theses & Dissertations

Biofilm production plays a crucial role in bacterial antibiotic tolerance and novel approaches are needed to combat it. The stringent response (SR), activated by nucleotide alarmones (pp)pGpp, is crucial for bacterial transcriptional reprogramming. In Clostridioides difficile, the SR is implicated in antibiotic survival, contributing to infection recurrence. Despite its known roles in sporulation and biofilm production in other bacterial species, the broader influence of the SR on C. difficile physiology remains undiscovered. This study deletes the C. difficile SR gene relQ, revealing the pivotal role of RelQ in regulating SR-dependent phenotypes in this organism. The absence of RelQ …

Pulsed Electric Fields Sensitize Methicillin-Resistant Staphylococcus Aureus To Antibacterial Therapies And Stimulate Host Immune Responses, Alexandra E. Chittams-Miles

Biomedical Sciences Theses & Dissertations

This research explores the impact of nanosecond pulsed electric fields (nsPEF) on two fronts: their immune stimulatory effects and their potential as a novel strategy to enhance the sensitivity of Methicillin-resistant Staphylococcus aureus (MRSA) to clinically relevant antibiotics. While pulsed electric fields have been reported to have an immune stimulatory effect, the mechanisms responsible for these effects have yet to be determined.

Our investigation addresses the rising concern of MRSA derived skin and soft tissue infections (SSTIs). Consistent with other publications, we found that nsPEF alone cause modest inactivation of planktonic MRSA. We then investigated the effects of nsPEF in …

Modified Vaccine Vectors To Understand Adjuvant Functions Of Listeria During Chronic Schistosomiasis, Stephanie K. Norwood

Biological Sciences Theses & Dissertations

Vaccination is one of the most effective strategies employed to prevent infectious diseases. Successful vaccination is dependent upon the induction of a specific, robust, and prolonged immune response. One of the major challenges faced by vaccine development is vaccine failure due to host-related factors that can modulate the immune system, which leads to non-responsiveness to vaccinations. The generation of new vaccine strategies is imperative to combat these effects. Live bacterial vectors are one approach used as they can elicit humoral immunity, cellular immunity, or both. Listeria monocytogenes is a Gram positive, intracellular pathogen that is an effective bacterial vaccine vector …

T-Cell Antigen Receptors In Multiple Sclerosis, Lisa Lanée Keyes Jones

Biomedical Sciences Theses & Dissertations

Multiple Sclerosis (MS) is T-cell mediated autoimmune disease characterized by inflammation, demyelination, and degeneration of axons in the brain and spinal cord. A T cell-mediated immune response in MS is directed against myelin components and possibly other antigens in genetically susceptible individuals and is triggered by a viral infection. The T-cell antigen receptor (TCR) on T cells is responsible for antigen recognition and determines specificity. Our overall hypothesis is to determine whether clonally expanded T cells in patients with MS recognize viral or self-antigens and to determine whether molecular mimicry is involved in the development of the disease. To study …

The Influences Of Disperser Behavior, Host Availability, And Environmental Conditions On The Distribution Of Oak Mistletoe [Phoradendron Leucarpum (Raf.) Reveal & M. C. Johnst.] In Eastern Virginia And North Carolina, Nicholas Pearce Flanders

Biological Sciences Theses & Dissertations

Mistletoes are shrubs that typically parasitize the branches of host trees and rely on avian frugivores for seed dispersal. Because mistletoes are restricted to a narrow range of suitable recruitment sites and avian frugivores are more visible than other guilds of seed dispersers, mistletoe-frugivore systems afford opportunities for assessing the roles of dispersal limitation and local environment in determining plant distribution. These mechanisms have been proposed as determinants of the observed association of oak mistletoe [Phoradendron leucarpum (Raf.) Reveal & M. C. Johnst.] with forested wetlands in eastern Virginia and North Carolina, USA. I tested the alternative hypothesis that …

The Impact Of Nanopulse Treatment On The Tumor Microenvironment In Breast Cancer: Overturning The Treg Immunosuppressive Dominance, Anthony Nanajian

Biomedical Sciences Theses & Dissertations

Nanopulse treatment (NPT) is a high-power electric engineering modality that has been shown to be an effective local tumor treatment approach in multiple cancer models. Our previous studies on the orthotopic 4T1-luc breast cancer model demonstrated that NPT ablated local tumors. The treatment consequently conferred protection against a second live tumor challenge and minimized spontaneous metastasis. This study aims to understand how NPT mounts a potent immune response in a predominantly immunosuppressive tumor.

NPT changed the local and systemic dynamics of immunosuppressive cells by significantly reducing the numbers of regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), and tumor-associated macrophages …

Role Of Ixodes Scapularis Sphingomyelinase-Like Protein (Issmase) In Tick Pathogen Interactions, Pravesh Regmi

Biological Sciences Theses & Dissertations

Arthropod-borne diseases are one of the major concerns throughout the world. Ixodes scapularis (hard tick) is one of the major vectors that is involved in arthropod-borne disease transmission. Langat virus (LGTV) is a model pathogen that is very similar to other medically important flaviviruses such as Tick-Borne Encephalitis virus (TBEV) and Powassan virus (POWV). Sphingomyelinase-like protein (IsSMase, a Sphingomyelinase D or SMase D, a venomous protein ortholog of spiders) is an enzyme present in ticks that helps to catalyze the hydrolysis of the sphingomyelin (cell membrane lipid) into phosphocholine and ceramide. The objective of our study is to delineate the …

Effects Of Carboxylated Nanodiamonds On Macrophages During And After Differentiation, Maisoun E. Bani Hani

Biological Sciences Theses & Dissertations

Nanodiamonds (ND) are a carbon-based nanomaterial that are increasingly being proposed for developing novel imaging techniques, as carriers of biomolecules and therapeutic drugs, as coatings for implants, and for other biomedical applications. The exceptional chemical, mechanical, and optical properties of ND make this material suitable in a wide range of fields. The application of ND in the biomedical field is attractive but requires more in-depth investigation into the safety of ND and its interactions with different cells and systems. The effects of ND on the immune system are not fully understood or investigated and there are several controverting reports regarding …

Abiotic Factors Contributing To The Survival Of Three Tick Species In Southeastern Virginia, Amblyomma Americanum (Lone Star Tick), Dermacentor Variabilis (American Dog Tick), And Amblyomma Maculatum (Gulf Coast Tick), Lindsey A. Bidder

Biological Sciences Theses & Dissertations

Amblyomma americanum, Amblyomma maculatum, and Dermacentor variabilis are hard-bodied ticks in the Hampton Roads area of southeastern Virginia. This study consisted of two field projects focused on these tick species. To estimate the off-host survival of local tick species, a capture-mark-recapture (CMR) study was performed. An environmental survival study was performed to quantify the ability of these three tick species to survive in situ. Four field sites were used in the Hampton Roads region covering a variety of habitat types and vegetation; specifically two drier, upland field sites and two flood-prone sites. CMR was conducted from May through …

A Study Of A Simulated Infectious Disease On Healthcare Workers' Reaction, Knowledge, Attitudes, And Performance Towards Hand Hygiene, Lydia Wigglesworth-Ballard

Health Services Research Dissertations

Problem Statement. The high rate of healthcare worker-patient contact provides many opportunities for exposure to pathogens, which creates challenges in assessing healthcare workers' success at preventing healthcare associated infections caused by these dangerous pathogens.

Methods. This study explored the effects of a simulated infectious disease on healthcare workers' hand hygiene knowledge, attitudes, performance, and reactions using Kirkpatrick's Four levels of Evaluation. The study utilized a mixed method pre-test – 2 post-test design. The dependent variables were hand hygiene knowledge, attitudes toward hand hygiene guidelines, hand hygiene performance, and reaction to the overall experience. Data was collected three times over a …

Discretized Agent-Based Model Of Infectious Disease Spread That Uses Contact Probability, Tyrell L. Gardner

Computational Modeling & Simulation Engineering Theses & Dissertations

This study uses contact probability in an agent-based model to simulate the spread of an infectious disease. In order to perform the study, the agent-based model must first be discretized into events. Each agent in the model is given its own infectious disease state machine taken from the Susceptible-Exposed-Infected-Recovered (SEIR) model. The agents move between squares in a grid environment where each square represents a group. Groups have a contact probability as an attribute that is used to predict whether an agent comes in close contact with another agent. The transitions between the states in the SEIR model are easily …

Ecological Interactions Between Spiders And The Purple Pitcher Plant, Sarracenia Purpurea, Marc Aaron Milne

Biological Sciences Theses & Dissertations

Spiders and harvestmen are commonly captured by or reside upon the carnivorous purple pitcher plant, Sarracenia purpurea. Although spiders and harvestmen are often known to be prey of S. purpurea, other ecological interactions between these arthropods and the plant are poorly understood. Studies were undertaken at three pitcher plant populations, two in Virginia and one in North Carolina, to assess the ecological relationships between spiders and harvestmen and S. purpurea. Multiple plots containing pitcher plants (treatment) and plots lacking pitcher plants (control) were created at these locations. Spiders and harvestmen were collected through five different techniques. Spatial, …

Lipid Targets Of The Antimalarial Trioxanes In Plasmodium Falciparum, Carmony Leah Hartwig

Biological Sciences Theses & Dissertations

Malaria is among the most debilitating diseases of man. The protozoan parasite, Plasmodium falciparum, causes over a million annual fatalities. The antimalarial trioxanes, exemplified by artemisinin, are among the few pharmaceuticals for which clinical resistance has not become widespread. Artemisinin is a naturally occurring sesquiterpene lactone, containing a unique endoperoxide pharmacophore. Despite extensive study, the precise antimalarial mechanism of action of trioxanes remains elusive. Heme iron-mediated cleavage of the endoperoxide within the parasite digestive vacuole is hypothesized to generate cytotoxic metabolites capable of alkylating heme and damaging cellular macromolecules. The hypothesis of this research is that the endoperoxide pharmacophore …

Comparison Of Immune Correlates, Age Related Changes And Proteomic Profiling Of Healthy Individuals Receiving Influenza Vaccines, Gaurav Basu

Theses and Dissertations in Biomedical Sciences

The burden of influenza related infections is substantial, both in terms of illness, lives lost and economic impact on society. The degree of impact of influenza related infections is much higher in the elderly population where it is a leading cause of catastrophic disability; greatly affecting the quality of life of elderly persons above 65 years of age. Vaccination is the mainstay for control and prevention of influenza infections. There are two vaccine formulations that are licensed for use at present. The inactivated influenza vaccines (TIIV) which have been used for 60 years in all age groups and the new …

The Plasmodium Falciparum Chloroquine Resistance Transporter, Pfcrt, Mediates The Activity Of Chloroquine-Resistance Reversal Agents In The Malaria Parasite, Kristin Lane

Biological Sciences Theses & Dissertations

Chloroquine (CQ) resistant Plasmodium falciparum is a serious problem affecting 3.2 billion people in over 100 countries today. Most endemic malarious countries are among the poorest in the world and lack the resources to replace the inexpensive and highly effective CQ. CQ resistance (CQR) reversal agents are a potentially inexpensive solution to restoring CQ efficacy. CQR reversal agents are drugs that have little to no antimalarial activity alone, but in combination with CQ, they increase dmg accumulation in the parasite and enhance the sensitivity to CQ in CQR parasites. PfCRT is a putative transporter located on the parasite digestive vacuole …

Down-Regulation Of Natural Killer Cell Activation In Response To Influenza Virus In Older Adults, Yu Jing

Theses and Dissertations in Biomedical Sciences

Immune senescence contributes to influenza-associated high mortality and morbidity and reduced vaccine efficacy in elderly people. Type I T cell (Thl)-mediated immunity plays a significant role in Immune responses to influenza infection and vaccination. Natural killer (NK) cells secrete significant amount of IFN-7 , a hallmark Thl cytokine, in response to influenza infection. How aging influences human NK cell IFN-7 production in response to influenza virus has not been well documented. In this study we employed human peripheral blood mononuclear cells (PBMC) and performed intracellular cytokine staining and flow cytometry primarily to investigate how aging influences NK cell activation with …

The Defensin Gene In Dermacentor Variabilis And Ixodes Scapularis, Katherine Corbett Seguin

Biological Sciences Theses & Dissertations

In response to microbial challenge, invertebrates rely on their innate immune response to recognize and destroy harmful pathogens. One of the primary proteins produced by the response and that has been observed in many invertebrates, including insects, mussels, and ticks, is defensin. These proteins are responsible for forming pores in the membranes of invading microbes thereby destroying them. This study examined the gene coding region for defensin in two important tick disease vectors, Dermacentor variabilis and Ixodes scapularis. PCR amplification of the genes from tick chromosomal DNA resulted in the amplification of a 225 base pair amplicon. Sequencing of …

Characterization Of Mouse Prostate Tumor-Infiltrating Leukocytes, Daniel Alan Holterman

Theses and Dissertations in Biomedical Sciences

Prostate cancer is the most common form of non-cutaneous cancer among men in the United States and the second most common cause of cancer-related death among men with approximately 189,000 new cases diagnosed and 30,200 deaths of the disease in 2002. Prostate cancer can be a treatable disease but once it becomes metastatic there are no acceptable therapies. For this reason, immunotherapy has been attempted but the results have been disappointing. The TRAMP model was used to evaluate specific interactions between an intact murine immune system and a prostate tumor that expressed a naturally processed weak tumor antigen. Vaccination trials …

Investigations Into The Innate Resistance Of The Tick, Dermacentor Variabilis, When Challenged With The Bacterium, Escherichia Coli, Shane Michael Ceraul

Biological Sciences Theses & Dissertations

In addition to the soluble response, many invertebrates control bacterial infections by means of phagocytosis or melanotic encapsulation. In some insects, Escherichia coli growth is reported to be inhibited by aggregation/encapsulation. Soluble and phagocytic responses to bacterial challenge have been reported in ticks, but evidence of an aggregation/encapsulation response was reported only for inanimate (araldite) implants. This study was done to determine how ticks control infection by E. coli. Ticks were challenged by direct inoculation of bacteria into the hemocoel cavity. Using plate counts, no viable E. coli were detected I hour post-inoculation. A direct fluorescence assay (DF A) …

Diffusion Problems In Wound Healing And A Scattering Approach To Immune System Interactions, Julia Suzanne Arnold

Mathematics & Statistics Theses & Dissertations

A theoretical model for the existence of a Critical Size Defect (CSD) in certain animals is the focus of the majority of this dissertation. Adam [1] recently developed a one-dimensional model of this phenomenon, and chapters I–V address the exist the CSD in a two-dimensional model and a three-dimensional model. The two dimensional (or 1-d circular) model is the more appropriate for a study of CSD's. In that model we assume a circular wound of uniform depth and develop a time-independent form of the diffusion equation relevant to the study of the CSD phenomenon. It transpires that the range of …

Cytotoxic T Cell Response To Influenza Vaccination In Older Adults, Caroline R. Letter

Chemistry & Biochemistry Theses & Dissertations

Influenza infections result in activation of cellular and humoral immune responses, leading to stimulation of cytotoxic T cells (CTL) and helper T cells with subsequent viral clearance.

By using an assay of granzyme B activity, the in vivo cellular-mediated immune response of old adults was found to be significantly decreased in comparison to young adults. The effect of aging on the CTL response was further investigated via enzyme-linked immunospot technique. A significant increase in number of virus-specific T cells occurred in older adults after vaccination, although the magnitude of the CTL response decreased in comparison to young adults. These data …

The Effect Of Illicit Drugs On The Cardiovascular And Hormonal Responses In Pregnant Rabbits, Andrea Alana Chambliss

Biological Sciences Theses & Dissertations

The purpose of this study is to determine the effect of acute administration of heroin and ecgonine methyl ester (EME) on renin and atrial natriuretic peptide (ANP) secretion in time dated New Zealand White rabbits. Thirteen rabbits were studied. Arterial blood pressure and pCOz, increased significantly from a control value of 80 ± 8 mmHg and 30 ± 2 mmHg respectively to 93 ± 18 mmHg and 31 ± 1 mmHg. pO₂ decreased from 99 ± 8 mmHg to 90 ± 11 mmHg 10 minutes after heroin administration. EME caused arterial blood pressure to significantly increase from a control …

Characterization Of P39, A Borrelia Burgdorferi Specific Protein, Jill M. Troyer

Theses and Dissertations in Biomedical Sciences

Borrelia burgdorferi, the causative agent of Lyme disease, expresses a species specific, 39 kiloDalton protein of unknown function called P39. This protein is highly immunogenic and the presence of anti-P39 antibodies in patient sera is used as an indicator of B. burgdorferi infection. P39 is also a candidate for vaccine developement due to conserved expression of P39 among B. burgdorferi isolates. Since little is presently known about this important molecule, the present study was designed to characterize P39 and was carried out in six separate investigations. First, infectious and noninfectious isolates of B. burgdorferi were established in vitro, and …

Neuronal Toxicity Of Humoral Factors From Patients With Diabetic Neuropathy And Mechanism Of Neuronal Death On Cultured Adrenergic Neurons: Studies Of Autoimmune Pathogenesis In Diabetic Neuropathy, Dong Liu

Theses and Dissertations in Biomedical Sciences

Diabetic neuropathy is an extremely common medical problem because of the high frequency of diabetes and its complications. Diabetic neuropathy consists of not one, but many patterns of nerve injury, each potentially with its own discrete pathogenic mechanisms. Evidence is accumulating that indicates autoimmunity plays a role in diabetic peripheral neuropathy. However, the significance of immunopathogenesis of diabetic neuropathy is as yet unclear.

Using the N1E-115 mouse neuroblastoma cell line (NB) as a model of the adrenergic autonomic neuron, we found that sera of IDDM patients with neuropathy inhibit growth and differentiation of adrenergic neurons in culture (Pittenger et al, …

Characterization Of An Antiviral Agent Based On Nonionic Surfactants And It's Effects On Human Dermal Fibroblasts, Ji Young Li

Biological Sciences Theses & Dissertations

The AIDS ·pandemic has directed various research endeavors towards finding an appropriate method for eliminating all potentially infectious material within bone allografts before implantation into a recipient. To that effect, Panavirocide was conceived by Medicine and Applied Science·s, Inc. This compound, in which three nonionic surfactants serve as the active agents, has been shown to inactivate HIV-1 particles within blood and blood products. Because Panavirocide has never been used with allografts, the purpose of the present research was to characterize the nonionic surfactants for the putative alteration of the formulation by determining the critical micelle concentration values and to determine …

Purification And Characterization Of The Prostate Cancer-Associated Antigen: 7e11-C5, Qi Feng

Theses and Dissertations in Biomedical Sciences

A murine monoclonal antibody (MAb), 7E11-C5, recognizes a prostate organ-specific antigen. Under reducing conditions, the 7E11-C5 antigen exhibited major 100-Kd and minor 70-Kd bands, suggesting that the antigen comprises two peptide chains. The antigen was detected in pooled normal human seminal plasma, xenograft tissue extract of a prostate adenocarcinoma cell line, LNCaP, and tissue extracts of benign prostatic hyperplasia (BPH) and prostate carcinomas, but was not identified in any of the non-prostate tissue extracts tested. Antibodies of the three major prostate-associated antigens (prostatic acid phosphatase (PAP), prostate specific antigen (PSA) and prostate secretory protein (PSP)) were unable to block the …

Development And Characterization Of Monoclonal Antisperm Antibodies: Potential For Contraception, Dilrowshan H. Haque

Theses and Dissertations in Biomedical Sciences

There is an expressed need to develop a greater variety of safe contraceptive methods which would find acceptance worldwide, particularly in developing countries. Immunization against spermatozoa might be such a method as judged by the accumulated evidence from studies on human females and in a number of female animal species.

Two extraction techniques were used for sperm membrane antigen isolation. The first technique involved NP-40 detergent for antigen extraction from human motile sperm and the second technique employed homogenized human testis for antigen extraction. Using these immunogens, sperm membrane-specific monoclonal antibodies (MAbs) were developed. When these antisperm MAbs were subjected …

Evidence Of A Neurogenic Component During Ige-Mediated Inflammation In Murine Skin, Victoria Jean Cavanaugh

Biological Sciences Theses & Dissertations

The direct stimulation of sensory nerves or the intradermal injection of neuropeptides triggers a cutaneous inflammatory response that involves the degranulation of mast cells. These neurogenic inflammatory reactions are physiologically similar to IgE-mediated hypersensitivity responses, which also depend on mast cells. Although non-antigenic stimuli distinguish neurogenic from IgE-mediated inflammation, the similarity of their effector stages suggested that peripheral nerves may also participate in lgE-mediated responses. To examine this hypothesis, IgE responses were elicited in denervated skin. A murine model was developed where the footpad was denervated by surgically removing a 2 mm segment of the sciatic nerve, proximal to the …

Incorporation Of Carbon-14 Metabolites Into The Sex Pheromon 26-Dichlorophenol Of Dermacentor Andersoni Stiles; And The Disruption Of Pheromone Activity In D. Andersoni And D. Variabilis (Say) By Quinones, Daniel Martin Gainsburg

Biological Sciences Theses & Dissertations

The Rocky Mountain wood tick, Dermacentor andersoni Stiles, injected with (u-¹⁴c) sodium acetate or DL-(benzene ring-u-¹⁴c) tryptophan for in vivo studies, was found to produce sex pheromone, 2,6-dichlorophenol, containing ¹⁴c activity. Ambiguous results were obtained with L-(u- ¹⁴c) tyrosine, and no incorporation of ¹⁴c label into pheromone was observed with DL-(ring-u- ¹⁴c) phenylalanine or DL-(side chain-3-¹⁴c) tyrosine. Attempts were made to determine the site of synthesis, but in vitro studies proved inconclusive.

Disruption of sex pheromone activity in D. andersoni and the American dog tick, D. variabilis (Say) was achieved …

Detection Of Unique Tumor-Associated Proteins In The Urine Of Patients With Transitional Cell Carcinoma By High Resolution Two-Dimensional Polyacrylamide Gel Electrophoresis, Robert David Lehman

Biological Sciences Theses & Dissertations

A two-dimensional polyacrylamide gel electrophoretic technique was developed for the detection of unique tumor-associated proteins in urine of patients with transitional cell carcinoma (TCC) of the urinary bladder. This technique yielded high resolution of polypeptides in the urine of cancer patients. Urine was chosen as a source of tumor-associated components because the location of the tum or should release already solublized tum or substances in to the urine.

Urinary proteins were concentrated and then analyzed by the two - dimensional electrophoretic technique using isoelectric focusing in the first dimension and sodium dodecylsulfate - polyacrylamide gel electrophoresis in the second dimension. …