Life Sciences Commons^™

Investigating The Involvement Of The Tick Vector In The Induction Of Alpha-Galactose Hypersensitivity (Alpha-Gal Syndrome, Red Meat Allergy) In The United States., Gary Crispell

Dissertations

Alpha-gal syndrome (AGS or sometimes called red meat allergy) is a result of the development of specific IgE antibodies to the oligosaccharide galactose-α-1,3-galactose (α-gal) after a person has had exposure to tick bites. This dissertation investigates four common tick species found in North America: the lone-star tick (Amblyomma americanum), the Gulf-Coast tick (Amblyomma maculatum), the American dog tick (Dermacentor variabilis), and the black-legged tick (Ixodes scapularis) for the presence of α-gal by utilizing a combination of immunoproteomic approaches and carbohydrate analysis techniques.

Anti-α-gal IgM antibodies (M86) were used in immunoblotting to detect …

Characterizing Endogenous Dicer Products To Unravel Novel Rnai Biogenesis Pathways, Jacob Oche Peter

Dissertations

ABSTRACT

RNA interference (RNAi) is a pervasive gene regulatory mechanism in eukaryotes based on the action of multiple classes of small RNA (sRNA). Exploiting RNAi pathways in non-model systems have great potential for creating potent RNAi technologies. Here, we accessed RNAi-mediated control of gene expression in the two-spotted spider mite, Tetranychus urticae (T. urticae) using engineered dsRNA designed to modulate the host RNAi pathway and increase RNAi efficacy. Analysis of Dicer (Dcr) generated fragments revealed how exogenous RNAs access the host RNAi pathway in this animal, opening avenues for designing RNAi technology for their control. Further, some organisms …

Sensitivity Of A Lamp Assay For Detection Of The Dinoflagellate Amyloodinium Ocellatum In Simulated Field Conditions And Freeze Tolerance Of The Parasite, Robert Gonzales

Master's Theses

The obligate parasitic dinoflagellate Amyloodinium ocellatum causes amyloodiniosis in warm water marine fishes. The prolific parasite, which has a direct, three-stage life cycle, is highly infectious and can cause heavy losses in aquaculture. Prevention, biosecurity, and early detection are vital for control. In this work, microscopy and a loop-mediated isothermal amplification (LAMP) assay were compared for early diagnosis of A. ocellatum in cultured stocks, and the freeze tolerance of tomonts was assessed to determine if frozen wild fish used as fish food can serve as a potential vector for the parasite.

The lowest dinospore concentration that could be detected by …

Dicer Represses Antiviral Innate Immunity Pathways In Mouse Embryonic Stem Cells, Chandan Gurung

Dissertations

Recent studies have demonstrated that embryonic stem cells (ESCs) are deficient in expressing type I interferons (IFN), the cytokines that play key roles in antiviral responses. However, the underlying molecular mechanisms and biological implications of this finding are poorly understood. In this study, I used a synthetic RNA-based assay that can simultaneously assess multiple forms of antiviral responses in ESCs. Dicer is an enzyme essential for RNA interference (RNAi), which is used as a major antiviral mechanism in invertebrates but not clear in vertebrates. RNAi activity is detected in wild-type ESCs but is abolished in Dicer knockout ESCs (D−/−ESCs) as …

Effects Of Infection Of The Protist Parasite, Dermomycoides Sp., In Dusky Gopher Frog Tadpoles, Jaime Smith

Master's Theses

Infections of the protist parasite, Dermomycoides sp. are thought to have caused several years of low recruitment in the dusky gopher frog (Rana sevosa) populations. I evaluated the effects of density of the infective zoospores, host developmental stage, and tadpoles' ability to acquire resistance to Dermomycoides sp. on dusky gopher frog tadpoles. Tadpoles were exposed to zoospore densities of 0, 250, 500, and 750 zoospores/µL at Gosner stage 25, and we found no significant differences among treatments in tadpole mortality. In evaluating susceptibility by development stage, I exposed R. sevosa to 50 zoospores/µL as eggs, embryos, hatchlings, and …

Generating Zika Vaccine Candidates Using Nhumirim Virus As A Backbone, Mariam A. Atobiloye

Honors Theses

Zika virus (ZIKV) is a mosquito-transmitted flavivirus that is causing significant world-wide health concern. There are currently no treatments or vaccines available for this virus, thus, there is an urgent need to develop a safe and effective vaccine to combat ZIKV infection. Nhumirim virus (NHUV) is also a mosquito-transmitted flavivirus, but it is unable to infect humans and other vertebrate animals, making it an ideal candidate to develop chimeric viral vaccines against other disease-causing flaviviruses, such as ZIKV. In this study, we generated chimeric viruses by replacing envelope (E) gene in the genome of NHUV with ZIKV E gene, which …

Characterizing Erad And Antioxidant Response In Ixodes Scapularis Ise6 Cells During Borrelia Infection, Latoyia Downs

Master's Theses

Ticks are hematophagous ectoparasites that transmit a multitude of diseases to humans. Borrelia burgdorferi (BB) and Borrelia miyamotoi (BM) are both tick-borne pathogens that cause disease in humans and are transmitted by the black-legged tick (Ixodes scapularis). A byproduct of blood digestion generates reactive oxygen species that are toxic and cause oxidative stress which promotes cellular damage and dysfunction. The endoplasmic reticulum (ER) is especially affected by oxidative stress, resulting in a buildup of improperly folded proteins in the ER lumen called ER stress. To prevent cellular damage, the tick utilizes an antioxidant system to neutralize ROS and mechanisms …

Development Of An Attenuated Zika Virus By Editing The 5’ Untranslated Region, E. Ashley Thompson

Master's Theses

Zika Virus (ZIKV) is a mosquito-transmitted flavivirus that usually causes no symptoms to mild febrile in humans, and it has been regarded as an insignificant pathogen to public health. However, recent outbreaks of ZIKV infection have revealed that ZIKV can cause severe neurological effects in adults, such as Guillain-Barre Syndrome (GBS), and in infants whose mothers acquired the virus during pregnancy, causing Congenital Zika Syndrome (CZS). Currently, no approved vaccine is available and there is a critical need to develop an effective and safe vaccine. While most vaccine developmental strategies target the viral prM-E protein of ZIKV, we aimed to …

The Histoplasma Capsulatum Ddr48 Gene Is Required For Survival Within Macrophages And Resistance To Oxidative Stress And Antifungal Drugs, Logan Blancett

Dissertations

Histoplasma capsulatum(Hc)is a systemic, dimorphic fungal pathogen that affects upwards of 500,000 individuals in the United States annually. Hc grows as a multicellular mold at environmental temperatures; whereas, upon inhalation into a human or other mammalian host, it transforms into a unicellular, pathogenic yeast. The research presented in this dissertation is focused on characterizing the DNA damage-responsive gene HcDDR48. HcDDR48was originally isolated via a subtractive DNA library enriched for transcripts enriched in the mold-phase of Hcgrowth. Upon further analysis we found that HcDDR48is not just expressed in the mold morphotype, but both growth programs …

Attenuated Responses To Inflammatory Cytokines In Mouse Embryonic Stem Cells: Biological Implications And The Molecular Basis, Bohan Chen

Master's Theses

Embryonic stem cells (ESCs) have attracted intense interest due to their great potential for regenerative medicine. However, their immune property is an overlooked but a significant issue that needs to be thoroughly investigated not only to resolve the concern for therapeutic applications but also for further understanding the early stage of organismal development. Recent studies demonstrated that ESCs are deficient in innate immune responses to viral/bacterial infections and inflammatory cytokines. Inflammatory conditions generally inhibit cell proliferation, which could be detrimental to ESCs, since cell proliferation is their dedicated task during early embryogenesis. Thus, I hypothesize that the attenuated innate immunity …

Taxonomy And Systematics Of Plagioporus (Trematoda), With Descriptions Of 10 New Species From Freshwater Fishes Of The Nearctic, Thomas John Fayton

Dissertations

The Opecoelidae Ozaki, 1925 is one of the largest families of digenetic trematodes of fishes. While the family is mostly marine/estuarine, invasion of freshwater hosts has occurred at least two times. The only representative freshwater plagioporine sequenced to date is Plagiocirrus loboides Curran, Overstreet, & Tkach, 2007, which previous phylogenetic analyses resolved as being related to deep water marine opecoelids. The taxonomy of the freshwater plagioporines, particularly Plagioporus, has long been confused; homoplasy is rife within the family and has complicated the delineation of species and genera, and the freshwater species from marine forms. Here, I hypothesize that the …

A Novel Role For Osteopontin In Facilitating West Nile Virus Neuroinvasion, Amber M. Paul

Dissertations

West Nile virus (WNV) is a positive-sensed, single-stranded RNA flavivirus that can cause human neuroinvasive diseases, including encephalitis, meningitis, and flaccid paralysis. The mechanisms by which WNV enters the central nervous system and the host-factors that are involved in WNV-neuroinvasiveness are not completely understood. Osteopontin (OPN), a multifunctional glycoprotein, has been implicated as a bio-marker for a number of neuroinflammatory diseases. In particular, secreted (s)OPN has been implicated to participate in recruitment of polymorphonuclear neutrophils (PMN) to sites of its expression, while PMNs have been suggested to act as WNV reservoirs. Therefore, sOPN recruitment of PMNs may contribute to neuroinvasive …

Elucidating The Role Of Interleukin-17a In West Nile Virus Infection, Dhiraj Acharya

Dissertations

West Nile virus (WNV) is a neurotropic flavivirus of significant public health importance for which no therapeutics and vaccine are currently available. Interleukin-17A (IL-17A) is an inflammatory cytokine that regulates diverse immune functions, while its role is unclear in host’s immune response to WNV. Furthermore, CD8⁺ T cells are crucial components of immunity and play a vital role in recovery from WNV infection. Here, we report a previously unrecognized function of IL-17A in regulating CD8⁺ T cell cytotoxicity. We show that WNV induces the expression of IL-17A in both mouse splenocytes and human peripheral blood mononuclear cells cultured …

Study Of Rickettsia Parkeri Colonization And Proliferation In The Tick Host Amblyomma Maculatum (Acari: Ixodidae), Khemraj Budachetri

Dissertations

Amblyomma maculatum (Gulf coast tick) ticks are prevalent across the Atlantic to Gulf Coast region of United States. These ticks are recognized vectors of Rickettsia parkeri, a spotted fever group of Rickettsia (SFGR) known to cause American boutonneuse fever associated with fever and eschar rashes localized to the site of bites. We hypothesized that Rickettsia parkeri colonization and proliferation in the tick vector involve pathogen-symbiont dynamics and tick-pathogen interactions, which influence rickettsial transmission to the victims after tick bites. The rickettsial infection is maintained across the tick life cycle for many generations due to transovarial and transstadial transmission of …

A Microbiomic Approach To The Characterization Of The Impacts And Influences Of Viral, Bacterial, And Harmful Algal Bloom Toxins On The Bottlenose Dolphin, Corey David Russo

Dissertations

As apex predators that display high site fidelity Tursiops truncatus (bottlenose dolphin) are indicators of marine ecosystem health. Bottlenose dolphins, additionally, display pathogenesis and immune response similar to that of humans. Humans and coastal bottlenose dolphins, in particular, are constantly exposed to the same industrial, agricultural and domestic toxins and pathogens, contaminants and pollutants. Thus, studies on the bottlenose dolphin are also valuable in bridging the gap between ocean and human health. Bottlenose dolphins are susceptible to viral bacterial and toxin infection. Infection in the bottlenose dolphin manifests itself in the form of mass stranding events, unusual mortality events, chronic …

Antiviral Responses In Mouse Embryonic Stem Cells: Differential Development Of Cellular Mechanisms In Type I Interferon Production And Response, Ruoxing Wang

Dissertations

Embryonic stem cells (ESCs) have been recognized as a promising cell source for regenerative medicine. Intensive research over the past decade has led to the possibility that ESC-derived cells will be used for the treatment of human diseases. However, increasing evidence indicates that ESC-derived cells generated by the current differentiation methods are not fully functional. It is recently recognized that ESC-derived cells lack innate immunity to a wide range of infectious agents and inflammatory cytokines. When used in patients, ESC-derived cells would be placed in wounded sites that are exposed to various pathogens and inflammatory cytokines; therefore, their viability and …

Role Of Msa In Immune Evasion, Persistence, And Protease Regulation In The Human Pathogenic Strains Of Staphylococcus Aureus, Maria Deepa Basco

Dissertations

Opportunistic pathogens like Staphylococcus aureus on entering the host can stay colonized at the foci of infection or evade the immune system to disseminate to other sites. In this study we investigated the regulatory influence of the modulator of sarA (msa) on immune evasion and host persistence, employing the hospital-acquired strain S. aureus UAMS-1 and community-acquired strain S. aureus USA300 LAC. In the murine sepsis model, mutation of the msa gene in LAC showed no change in dissemination of infection; however, in UAMS-1 a decrease in microbial load was observed in the lungs. Differential regulation by the msa gene was …

Clam (Corbicula Fluminea) As A Potential Sentinel Of Human Norovirus Contamination In Freshwater, Xunyan Ye

Dissertations

The purpose of this study was to evaluate and validate the use of the clam Corbicula fluminea as a sentinel of human noroviruses (HuNoV) contamination in freshwater. The first specific aim was to develop a new method to extract HuNoV RNA from contaminated bivalves (e.g. oysters, clams) that would be much faster than existing methods. The procedure developed includes an initial total RNA extraction using TRI Reagent, followed by HuNoV RNA concentration and purification using biotinylated probe-capture technology. HuNoV RNA is finally detected by real-time RT-PCR. Using bivalve homogenates spiked with HuNoV, 100 PCR detection units of the virus was …

Genetic Variation In Potentially Virulent Vibrio Parahaemolyticus From The Northern Gulf Of Mexico, Nicholas Felix Noriea Iii

Dissertations

Vibrio parahaemolyticus (V_p) is a gram-negative bacterium found naturally in marine and estuarine environments. V_p is found in oysters including those which are later consumed by the public. Sub-populations of potentially virulent Vp contain specific virulence factors and are relevant human pathogens capable of causing gastroenteritis, wound infection, and death. The tdh and trh genes, both encoding hemolysins, have been correlated with the majority of clinical V_p isolates but have not been shown to be the definitive virulence factors.

A total of 146 V_p isolates from the northern Gulf of Mexico were collected and probed …

Influence Of Detritus Levels And Organic Pollution On Interspecific Resource Competition, Oviposition Behavior, And Larval Survival Of Two Tire-Inhabiting Mosquito Species (Diptera: Culicidae), David Wayne Allgood

Master's Theses

Mosquitoes (Diptera: Culicidae) are vectors of disease in the adult stage, but understanding the factors affecting distributions of the immature stages is important to the understanding and control of adult populations. Discarded automobile tires comprise important larval mosquito habitats. The Asian tiger mosquito (Aedes albopictus) and the southern house mosquito (Culex quinquefasciatus) are two medically important species commonly found in tires, but factors affecting their larval distributions in tires have not been studied, nor have their interspecific interactions. I investigated the effects of chemicals associated with organic pollution on oviposition preferences and larval survival of both …

Regulation Of Biofilm And Antibiotic-Resistance By The Modulator Of Sara (Msa) In Staphylococcus Aureus, Antony Schwartz

Dissertations

Staphylococcus aureus is an important human pathogen that is the causative agent of life-threatening diseases such as endocarditis and osteomyelitis. The ability of S. aureus to thrive as a successful pathogen can be partially attributed to its ability to form biofilm. Biofilm is an extracellular polysaccharide, protein, and DNA-based slime layer that protects the bacterial community. The global regulator sarA is essential for biofilm formation. Since the modulator of sarA (msa) gene regulates several virulence factors and is required for the full expression of sarA, the capacity of the msa mutant to form a biofilm was examined. The mutation of …

The Role Of Msa In The Global Regulation Of Virulence In Staphylococcus Aureus, Vijayaraj Nagarajan

Dissertations

Staphylococcus aureus is an important pathogen causing life threatening diseases in humans. Previously we showed that msa modulates the activity of sarA (Staphylococcal accessory regulator), which is one of a major global regulator of virulence in S. aureus. The objective of this study is to characterize the role of msa (Modulator of SarA) in the global regulation of virulence in S. aureus. Structure and function predictions were done using several computational tools and approaches to understand the nature of msa. A novel S. aureus microarray meta-database (SAMMD) was designed and developed to compare and contrast other transcriptomes with msa transcriptome. …