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Inferring a Network of Horizontal Gene Flow among Prokaryotes Using Complementary Approaches
Horizontal gene transfer (HGT), a mechanism that facilitates exchange of genetic material between organisms from different lineages, has a profound impact on prokaryotic evolution. To infer HGT, we first developed a comparative genomics-based tool, APP, which can perform phyletic pattern analysis using completely sequenced genomes to identify genes are unique to a genome or have sporadic distribution in its close relatives. Performance assessment against currently available tools on a manually created 18-genome dataset and 2 benchmarking datasets revealed the superior accuracy of APP over other methods. We then utilized a parametric method to construct a gene exchange network. The composition-based method, Jenson-Shannon Codon Bias (JS-CB), groups genes into clusters based on similar codon usage bias. These clusters were analyzed using APP and examined for the enrichment HGT associated marker genes, then annotated as of native or alien origin based on these multiple lines of evidence. Intergenome clustering enabled identification of genes mobilized across alien components of the genomes (alien-alien transfer) and from native components of donor genomes to the recipient genomes (native-alien transfer). Functional classification of alien gene clusters revealed that metabolism associated genes are most frequently mobilized, in concurrence with previous reports, and additionally, a large number of genes …
Date:
August 2022
Creator:
Sengupta, Soham
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Multi-Level Effects of Oxygen Exposure in Endothermic Insects
This dissertation examined the phenotypic plasticity of endothermic, flight and respiratory physiology in response to developmental oxygen exposure in the moth Manduca sexta. Development in both 10% O2 hypoxia and 30% O2 hyperoxia treatments were used to look at the physiological consequence on both ends of the oxygen spectrum. Hypoxic insects reached smaller sizes as adults and had longer pupation lengths than controls. Hyperoxic insects were larger at the end of the larval stage, had increased larval growth rates, but also had longer developmental larval developmental times and pupation lengths than controls. There was a decrease in both metabolic rate and thorax temperatures of hypoxic reared insects at normoxic levels. In flight trials hypoxic insects had the lowest critical flight PO2, and the hyperoxic insects had the highest PO2. There was an increase in hypoxic insect flight muscle mitochondria oxygen consumption in permeabilized fibers, but this did not translate to the isolated flight muscle mitochondria metabolic rates. Rearing oxygen level did not significantly affect mitochondrial density and size; myofibril density and size, or tracheal density and size in flight muscle. Overall, I found that higher levels of organization were more susceptible to the effects of chronic oxygen exposure and found …
Date:
August 2022
Creator:
Wilmsen, Sara M
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Regulation of Receptors in Neuronal Cilia with Development, Seizures, and Knockouts: Implications for Excitability
Neurons commonly have a primary cilium, which is a non-motile organelle extending from the centrosome into the extracellular space. In most brain regions, neuronal cilia are enriched in either somatostatin receptor type 3 (SstR3) or melanin concentrating hormone receptor type 1 (MCHR1), or both. The present immunohistochemical study provides novel evidence that primary cilia regulate neuronal excitability via G-protein coupled receptors (GPCRs), and that their identity is governed by brain region and by competition, both in adulthood and in postnatal development. The hippocampus, which is particularly vulnerable to seizures, has opposing gradients of SstR3(+) and MCHR1(+) ciliary GPCRs. We hypothesized that there is a competition between these two ciliary GPCRs, which might take place on any level from gene expression to presence in the cilium. We examined whether receptor colocalization occurs transiently in development before ciliary GPCR dominance is established in neurons in the CNS. In postnatal CA1 and CA3, the first GPCR to appear in cilia was the one that will dominate in adults: MCHR1 in CA1 and SstR3 in CA3. Some days later, the second GPCR was expressed along with the first; dual-receptor cilia were the exclusive type until single-receptor cilia emerged again around P14. Single-receptor cilia then …
Date:
August 2022
Creator:
Shrestha, Jessica
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Temperature Change and Its Consequences for the Physiology of the Eurythermic Sheepshead Minnow (Cyprinodon variegatus)
The estuarine sheepshead minnow (Cyprinodon variegatus) is the most eurythermic fish species, with a thermal tolerance window between 0.6°C and 45.1°C. However, little is known about the physiological mechanisms that allow this species to survive this temperature range. In order to understand how sheepshead minnow physiology is affected by temperature acclimation and acute changes in temperature, I conducted research on this species using a multi-level approach. I began at the organismal level, and examined the effects of these temperature changes on the sheepshead minnow's metabolic rate and swimming performance. The next chapter investigated the effects of changing temperatures on cardiac function (i.e., tissue/organ specific effects). In the final chapter, I conducted research at the sub-cellular level, and determined how mitochondrial bioenergetics / function is impacted by changing temperatures. This research shows that while sheepshead minnows are able to sustain heart function and mitochondrial respiration over a broad range of temperatures; they also display a plastic temperature response which is associated with the downregulation of standard metabolic rate and cardiac remodeling to maintain force generation. Collectively, these physiological responses may contribute to the sheepshead minnow's ability to maintain physiological and organismal function across a large temperature range.
Date:
August 2022
Creator:
Reynolds, Amanda Caroline
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Analysis of Multipartite Bacterial Genomes Using Alignment-Free and Alignment-Based Pipelines
In this work, we have performed comparative evolutionary analysis, functional genomics analysis, and machine learning analysis to identify the molecular factors that discriminate between multipartite and unipartite bacteria, with the goal to decipher taxon-specific factors and those that are prevalent across the taxa underlying the these traits. We assessed the roles of evolutionary mechanisms, namely, horizontal gene transfer and gene gain, in driving the divergence of bacteria with single and multiple chromosomes. In addition, we performed functional genomic analysis to garner support for our findings from comparative evolutionary analysis. We found genes such as those encoding conserved hypothetical protein DR_A0179 and hypothetical protein DR_A0109 in Deinococcus radiodurans R1, and Putative phage phi-C31 gp36 major capsid-like protein and hypothetical protein RSP_3729 in Rhodobacter sphaeroides 2.4.1, which are located on accessory chromosomes in both bacteria and were not found in the inferred ancestral sequences, and on the primary chromosomes, as well as were not found in their closest relatives with single chromosome within the same clade. These genes emphasize the important potential roles of the secondary chromosomes in helping multipartite bacteria to adapt to specialized environments or conditions. In addition, we applied machine learning algorithms to predict multipartite genomes based on gene …
Date:
August 2021
Creator:
Almalki, Fatemah
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Analysis of the Accumulation, Toxic Effects, and Risk of Persistent Organic Pollutants in Pinnipeds
The present studies determine the accumulation of persistent organic pollutants (POPs) in three pinniped species, evaluate the relationship with relevant biomarkers of exposure, and calculate toxic effect thresholds. Stranded harp and hooded seals were found to be accumulating PBDEs at levels which could pose a based on threshold levels determined in this study. Northern fur seals are accumulating all three classes of POPs (PCBs, PBDEs, and OCPs) with significant relationships being seen with blubber percent lipid. Correlations between contaminant concentrations and expression levels of relevant biomarkers were seen potentially indicating an effect on multiple pathways. Overall risk can be hard to determine due to factors such as sex and age. Broad threshold response values and hazard quotients were calculated for toxic effect endpoints in pinnipeds. Overall these results suggest that certain populations of pinnipeds are at high risk of experiencing toxic effects due to POP exposure, but it is important to understand effects even at lower concentrations. The relationship between exposure, toxic effects, and other stressors, both environmental and physiological, can impact the overall fitness and survival of pinnipeds.
Date:
August 2021
Creator:
Soulen, Brianne K
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Flow-Recruitment Relationships of Smallmouth Buffalo (Ictiobus bubalus) in Three Texas River Basins
This project focused on the relationship between instream flows and smallmouth buffalo (Ictiobus bubalus) recruitment in the Gulf Coastal Plain of Texas. The flow regime is the dominant factor in lotic systems and, consequently, the relationship between instream flows, including impacts to natural flow regimes, and life-history is a subject of growing interest. Smallmouth buffalo is a good model to investigate the relationship between river flows and variable interannual recruitment success of periodic life-history strategist fish species. Smallmouth buffalo were collected from the Brazos, Colorado, and Guadalupe Rivers of Texas, U.S.A., and otoliths were extracted from individuals in the field and sectioned and photographed in the lab. Photographs of sectioned otoliths were used to estimate age and thus the year in which the individual was spawned by counting back from the time of capture. Population age structure (i.e. a ‘state' or condition at a point in time) was used to infer effects of flow variation on a rates-based process (i.e. recruitment). After controlling for mortality using recruitment index values, interannual variation in recruitment was modeled using multiple components of the flow regime quantified as indicators of hydrologic alteration (IHA) variables based on daily discharge data from USGS gaging stations in …
Date:
August 2021
Creator:
Reeves, Cole Griffin
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Sociality in Harris's Hawks Revisited: Patterns of Reproductive Output and Delayed Dispersal
In the lower Rio Grande Valley of south Texas, more than half the nesting groups of Harris's hawks (Parabuteo unicinctus) include at least one auxiliary group member in addition to a breeding pair. To provide further insight into cooperatively breeding raptors, I evaluated sociality in Harris's hawks through the dual benefits framework. I explored the formation, structure, and stability of cooperative group formation across a spatially variable study area, which includes high levels of urbanization and development as well as remote, undisturbed native habitats with low anthropogenic impact. I used color banding, regular censuses of active territories, and a microsatellite relatedness analysis to examine patterns of sociality, including delayed dispersal, the effect of auxiliary group members on reproductive output, parentage of broods, and the relatedness of auxiliaries compared to the nestlings in their territories. I confirmed cooperative polygamy with genetic techniques for the first time in Harris's hawks and found 58% of juvenile hawks delayed dispersal for at least 6 mo. Using the dual benefits framework, I found social associations that formed through delayed dispersal followed predictions for resource-defense benefits, but sociality among mature non-related hawks more closely followed predictions associated with collective action benefits, specifically reproductive output was significantly …
Date:
August 2021
Creator:
Gibbons, Andrea L
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Investigating the Mechanisms involved in Traffic-Generated Air Pollution: Mediated Disruption of the Blood-Brain Barrier in a Wild Type Mouse Model using a Pharmaceutical Intervention Approach
This study investigated whether oxLDL and/or angiotensin (Ang) II signaling pathways mediate traffic-generated air pollution- exposure induced alterations in blood-brain barrier (BBB) integrity and permeability in a healthy wild type (C57Bl/6) mouse model; additionally, whether these outcomes are exacerbated by a high fat-diet investigated. An environmentally relevant concentration of a mixture of vehicle engine exhaust (MVE) was used. To investigate the hypotheses, 12 wk old male C57Bl/6 mice on either a high fat (HF) or low fat (LF) diet were randomly assigned to inhalational exposure of either filtered-air (FA) or 30 µg PM/m3 diesel exhaust + 70 µg PM/m3 gasoline exhaust (MVE) for 6 hr/day for 30 days. Additionally, we examined mechanisms involved in MVE-mediated alterations BBB integrity using a novel BBB co-culture in vitro model, consisting of mouse primary cerebral vascular endothelial cells on an apical transwell and astrocytes in the basal compartment, which was treated with plasma from the mice on our exposure study. Our in vivo exposure study results showed that MVE inhalation resulted in increased circulating plasma oxLDL and Ang II, compared to FA controls. Additionally, we observed increased cerebral microvascular expression of oxLDL receptors, LOX-1 and CD-36, and Ang II receptor subtype 1 (AT1) in …
Date:
August 2020
Creator:
Suwannasual, Usa
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Metabolic Responses to Crude Oil during Very Early Development in the Zebrafish (Danio rerio)
The present study sought to determine some morphological and physiological critical windows during very early development in zebrafish exposed to crude oil. I hypothesized that exposed zebrafish would present a decrease in survival rate and body mass, and an increase in routine oxygen consumption (ṀO2), and critical oxygen tension (PCrit). To test these hypotheses, zebrafish were acutely exposed (24 h) during different days of development (1 to 6 days post-fertilization, dpf) to different concentrations of high-energy water-accommodated fractions (HEWAFs). The endpoints of survival, body mass, routine oxygen consumption, and critical oxygen partial pressure were measured at 7 dpf. Survival rate decreased based on the exposure concentration but not as a function of the day of crude oil exposure. No significant effects were found in PCrit. Body mass was reduced by the different concentrations of HEWAF, with the size of the effect varying with exposure day, with the effect strongest on when exposure occurred at 2 and 3 dpf. Oxygen consumption (ṀO2) differed significantly depending upon the day of exposure in fish exposed to crude oil. Specifically, HEWAF exposure significantly increased ṀO2 in larvae exposed at 3 dpf (9.081 µmol O2/g/h, ±0.559) versus 2 dpf (6.068 µmol O2/g/h, ±0.652) and 6 …
Date:
August 2020
Creator:
Vazquez Roman, Karem Nathalie
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Ceramide Biosynthesis and NEET Proteins Impact Development, Function, and Maintenance of the Caenorhabditis elegans Germline
Access:
Use of this item is restricted to the UNT Community
I used the C. elegans genetic model to examine the role of ceramide biosynthesis (sphingolipid pathway) and iron regulation and found that each process impacts germline development and function. Using a sphingolipid specific antibody mAb15B4, I found that sphingolipids are associated with germ granules (P granules) within C. elegans and zebrafish; thus, suggesting conservation of macromolecules associated with germ granules. Phenotype analysis of ceramide biosynthesis mutants in C. elegans revealed that this pathway is essential for normal germline function in the aging adult hermaphrodite; specifically, precocious germline senescence was observed. Furthermore, I found that disruption of ceramide biosynthesis, via the hyl-2 deletion mutation, negatively impacts mAb15B4 localization at the P granules. Through genetic suppression analysis, I determined that insulin signaling and lipid biosynthesis can modulate the mAb15B4 localization to P granules. Additional, phenotype analysis showed that ceramide biosynthesis dysfunction decreased fecundity, and led to germline structure defects and uterine tumors. Through suppression analysis, I determined that modulation of the insulin signaling pathway suppressed the precocious germline senescence due to ceramide biosynthesis dysfunction. Since the presence of uterine tumors is associated with reproductive senescence I concluded that ceramide biosynthesis has a role in germline maintenance in the aging of the germline …
Date:
August 2019
Creator:
King, Skylar Dawn
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
The Metabolic Physiology of Planarian Flatworms
Access:
Use of this item is restricted to the UNT Community
Using a high throughput closed respirometry method to measure oxygen consumption, I determined metabolic rates in asexual and sexual Schmidtea mediterranea and Girardia dorotocephala, as a function of temperature, taxon, stressors, reproductive mode, age, regeneration, and specific dynamic action. This study has shown that oxygen consumption can reliably be measured in planaria using optode closed respirometry, and also provided a reliable method for measuring wet mass in planaria, which has been a challenge to researchers in the past. This research revealed that oxygen consumption in S. mediterranea is 1.5-2.1X greater in the sexual strain over the asexual strain at 13-18°C. Within the sexual strain, oxygen consumption is 1.5 -2.2X greater in sexually mature adults over the sexually immature groups (hatchlings, juveniles, and regenerating sexuals). Furthermore, I was able to quantify differences in sexual morphology between these groups exhibiting significant differences in oxygen consumption. The results of this research supports a theory of higher metabolic costs with sexual maturity in S. mediterranea. Therefore, this study has established sexual and asexual S. mediterranea as simple, yet attractive models for investigating energetic costs between sexual and asexual phenotypes. This research also provided quantitative values for specific dynamic action in planaria, with a maximum …
Date:
August 2019
Creator:
Lewallen, Melissa A
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Effects of Deepwater Horizon Crude Oil on Visual Function in Teleost Fishes
The Deepwater Horizon oil spill released millions of barrels of oil into the Gulf of Mexico, impacting economically and ecologically important fishes. Polycyclic aromatic hydrocarbons (PAHs) present in the oil have been shown to cause developmental impairments in early life stage fishes, such as morphological and behavioral changes related to eye formation and visual processing following PAH exposure. Prior research reported reduced eye growth in open water, pelagic species, as well as reduced photoreceptor-specific transcription factors associated with eye development following exposure to crude oil. Though changes in transcriptomic-level pathways associated with vision and visual processing have been reported, it has yet to be determined how these changes relate to physiological or behavioral-level effects in fish. Therefore, the present studies evaluated the effect of weathered crude oil on eye development and visual function in mahi-mahi, red drum, sheepshead minnow, and zebrafish larvae. Fish were assessed through several visually-mediated behavioral assays, analyzed histologically and immunohistologically, along with subsequent transcriptomic analyses and associated gene expression changes. Larvae exposed to crude oil experienced significantly reduced abilities to exhibit optomotor or optokinetic responses relative to controls, with associated reductions in retinal development. Furthermore, genes associated with eye development and phototransduction were downregulated, with subsequent …
Date:
August 2018
Creator:
Magnuson, Jason T
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Exposure to Nanomaterials Results in Alterations of Inflammatory and Atherosclerotic Signaling Pathways in the Coronary Vasculature of Wildtype Rodents
Cardiovascular disease (CVD) is the leading cause of death for people of most ethnicities on a global scale, and countless research efforts on the pathology of CVD has been well-characterized over the years. However, advancement in modern technologies, such as nanotechnology, has generated environmental and occupational health concerns within the scientific community. Current investigation of nanotoxicity calls into question the negative effects nanomaterials may invoke from their environmental, commercial, and therapeutic usage. As a result, further research is needed to investigate and characterize the toxicological implications associated with nanomaterial-exposure and CVD. We investigated the toxicity of multi-walled carbon nanotubes (MWCNT) and titanium dioxide (TiO2), which are two prominently used nanomaterials that have been previously linked to upregulation of inflammatory and atherogenic factors. However, the mechanistic pathways involved in these nanomaterials mediating detrimental effects on the heart and/or coronary vasculature have not yet been fully determined. Thus, we utilized two different routes of exposure in rodent models to assess alterations in proinflammatory and proatherogenic signaling pathways, which are represented in contrast throughout the dissertation. In our MWCNT study, we used C57Bl/6 mice exposed to MWCNTs (1 mg/m3) or filtered air (FA-Controls), via inhalation, for 6 hr/d for 14d. Conversely, intravenous TiO2 …
Date:
August 2018
Creator:
Davis, Griffith M.
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Genomic Island Discovery through Enrichment of Statistical Modeling with Biological Information
Horizontal gene transfer enables acquisition and dissemination of novel traits including antibiotic resistance and virulence among bacteria. Frequently such traits are gained through the acquisition of clusters of functionally related genes, often referred to as genomic islands (GIs). Quantifying horizontal flow of GIs and assessing their contributions to the emergence and evolution of novel metabolic traits in bacterial organisms are central to understanding the evolution of bacteria in general and the evolution of pathogenicity and antibiotic resistance in particular, a focus of this dissertation study. Methods for GI detection have also evolved with advances in sequencing and bioinformatics, however, comprehensive assessment of these methods has been lacking. This motivated us to assess the performance of current methods for identifying islands on broad datasets of well-characterized bacterial genomes and synthetic genomes, and leverage this information to develop a novel approach that circumvents the limitations of the current state-of-the-art in GI detection. The main findings from our assessment studies were 1) the methods have complementary strengths, 2) a gene-clustering method utilizing codon usage bias as the discriminant criterion, namely, JS-CB, is most efficient in localizing genomic islands, specifically the well-studied SCCmec resistance island in methicillin resistant Staphylococcus aureus (MRSA) genomes, and 3) …
Date:
August 2018
Creator:
Jani, Mehul
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Implications of Diet in Cardiovascular Disease Risk: Postprandial Changes in Circulating Monocytes and Endotoxemia
It is well established that continual consumption of a diet high in fat leads to the development of chronic conditions such as obesity, cardio metabolic syndrome, and atherosclerosis that are associated with high incidence of cardiovascular disease. Recent studies have identified endotoxin-derived inflammation as a major diving force for the development of these conditions. Our laboratory has recently demonstrated that consumption of a single high-fat meal results in acute postprandial endotoxemia and alters monocyte cell surface adhesion molecule expression and scavenger receptor CD36 expression. These collective projects describe our efforts to understand the physiological significance of these postprandial changes and if supplementation with spore-based probiotics are able to provide any form of protection against these responses that are associated with the onset of atherogenesis.
Date:
August 2018
Creator:
Venable, Andrea Henning
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Resistance Exercise and Alcohol: Combined Effects on Physiology and Performance
Resistance exercise (RE) training is a well-known and effective method for promoting increases in muscle mass and strength. A single bout of RE induces physiological disturbances that require coordinated activation of the immune system and intramuscular signaling in order to return the tissue to homeostasis and adapt to the RE challenge. On the other hand, acute binge alcohol consumption can affect the immune response to an inflammatory challenge, intramuscular anabolic signaling, and muscle protein synthesis, and the effects of alcohol on these processes are opposite that of RE. Furthermore, individuals who report more frequent exercise also report a greater frequency of binge drinking. However, few investigations exist regarding the effects of binge alcohol consumed after a bout of RE on RE-induced physiological changes and performance recovery. Therefore, the overarching purpose of the investigations contained within this dissertation was to investigate the effect of alcohol consumed after RE on the RE-induced changes in mTOR pathway signaling, muscle protein synthesis, inflammatory capacity, strength recovery, and power recovery. Although RE increased mTOR pathway signaling and inflammatory capacity after exercise and reduced maximal strength and explosive power the day after exercise, we observed no effects of alcohol (1.09 g ethanol∙kg-1 lean body mass, designed …
Date:
August 2018
Creator:
Levitt, Danielle E.
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
The Role of Thyroid Hormone on the Development of Endothermy in White Leghorn Chickens (Gallus gallus)
As chickens hatch, there is a rapid change in their physiology and metabolism associated with attaining endothermy. It is thought that thyroid hormones (TH) play a major role in regulating developmental changes at hatching. In birds, TH regulates skeletal muscle growth, which has a direct impact on the chick's ability to thermoregulate via shivering thermogenesis. To better understand the role of TH in the timing of hatching, development of thermogenic capacity, and metabolic rate, we manipulated plasma TH levels in chicken embryos beginning at 85% development (day 17 of a 21 day incubation) with either thyroperoxidase inhibitor methimazole (MMI) or supplemental triiodothyronine (T3). After TH manipulation, we characterized O2 consumption and body temperature in the thermal neutral zone and during gradual cooling. Externally pipped embryos and 1 day post hatch (dph) chicks were cooled from 35 to 15°C. Manipulation of TH altered the timing of hatching, accelerating hatching under hyperthyroid conditions and decelerating hatching with hypothyroid conditions. Cohen's d revealed a large effect size on body temperature (Tb) of EP embryos of hypothyroid animals when compared to euthyroid animals in environmental temperatures of 32°C to 15°C, which was not seen in 1dph animals. Hyperthyroid EP animals were able to maintain …
Date:
August 2018
Creator:
Rippamonti, Jessica D.
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
The Role of Transmembrane Protein 59 in Thrombocyte Function and the Effect of MS-222 on Hemostasis in Zebrafish
Transmembrane protein 59 (tmem59) is a gene that encodes a protein involved in autophagy and apoptosis in human. A previous study in zebrafish showed that tmem59 mRNA was several folds higher in thrombocytes than those found in red blood cells (RBCs). Therefore, we hypothesized that tmem59 has a role in thrombocytes function. We injected a hybrid of control vivo-morpholino (cVMO) and tmem59 specific antisense standard oligonucleotide (tmem59SO) into adult zebrafish to knockdown tmem59.This piggyback knockdown approach resulted in fish that had more bleeding in gill bleeding assay than the control fish. The thrombocytes fromtmem59 knockdown zebrafish aggregated faster with ADP and collagen agonists. Also, the number of blood cells was reduced after the knockdown of tmem59. We also found the effects of MS-222 anesthesia on hemostasis and found that the bleeding was reduced yielding less blood and the blood cell counts increased probably due to vasoconstriction of the blood vessels. In summary, we found tmem59 is a negative regulator of hemostasis and inferred that anesthesia should be avoided in hemostasis studies.
Date:
August 2018
Creator:
Deebani, Afnan Omar M.
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
The Acute Toxic Effects of the Synthetic Cannabinoid, JWH-018 on the Cardiovascular and Neuroendocrine Systems in Ictalurus punctatus (Channel Catfish)
Cannabinoid (CB) receptors have been found in most vertebrates that have been studied. The location of various CB receptors in the body and brain are known, but their physiological functions are not fully understood. The effects CBs have on the cardiovascular system have been of growing interest in recent years. Increasing reports from emergency departments and law enforcement agencies detail acute cardiovascular and psychological effects from synthetic CB intoxication, such as JWH-018. This major health concern is substantiated by governmental agencies like the CDC and NIDA. This pilot study investigates the acute toxic effects of the synthetic CB, JWH-018, on the cardiovascular and neuroendocrine systems in Ictalurus punctatus (channel catfish). Research in organisms besides the traditional mammal models can provide new insights into CB function and physiology. Ictalurus punctatus lend multiple benefits as a model organism that permits researchers to investigate in vivo effects of both cardiovascular and neuroendocrine systems without much influence from traditional sampling methods, and further more provide ample size and tissue to perform specific cardiovascular experiments. Multiple methods were used to assess cardiovascular function and sympathetic nervous system activation. Two different doses, low (500 µg/kg) and high 1,500 µg/kg, of JWH-018 were evaluated in the study. …
Date:
August 2017
Creator:
Taylor, Dedric Esmond
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Delivery of CRISPR/Cas9 RNAs into Blood Cells of Zebrafish: Potential for Genome Editing in Somatic Cells
Factor VIII is a clotting factor found on the intrinsic side of the coagulation cascade. A mutation in the factor VIII gene causes the disease Hemophilia A, for which there is no cure. The most common treatment is administration of recombinant factor VIII. However, this can cause an immune response that renders the treatment ineffective in certain hemophilia patients. For this reason a new treatment, or cure, needs to be developed. Gene editing is one solution to correcting the factor VIII mutation. CRISPR/Cas9 mediated gene editing introduces a double stranded break in the genomic DNA. Where this break occurs repair mechanisms cause insertions and deletions, or if a template oligonucleotide can be provided point mutations could be introduced or corrected. However, to accomplish this goal for editing factor VIII mutations, a way to deliver the components of CRISPR/Cas9 into somatic cells is needed. In this study, I confirmed that the CRISPR/Cas9 system was able to create a mutation in the factor VIII gene in zebrafish. I also showed that the components of CRISPR/Cas9 could be piggybacked by vivo morpholino into a variety of blood cells. This study also confirmed that the vivo morpholino did not interfere with the gRNA binding …
Date:
August 2017
Creator:
Schneider, Sara Jane
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Developing a Phylogeny Based Machine Learning Algorithm for Metagenomics
Metagenomics is the study of the totality of the complete genetic elements discovered from a defined environment. Different from traditional microbiology study, which only analyzes a small percent of microbes that could survive in laboratory, metagenomics allows researchers to get entire genetic information from all the samples in the communities. So metagenomics enables understanding of the target environments and the hidden relationships between bacteria and diseases. In order to efficiently analyze the metagenomics data, cutting-edge technologies for analyzing the relationships among microbes and communities are required. To overcome the challenges brought by rapid growth in metagenomics datasets, advances in novel methodologies for interpreting metagenomics data are clearly needed. The first two chapters of this dissertation summarize and compare the widely-used methods in metagenomics and integrate these methods into pipelines. Properly analyzing metagenomics data requires a variety of bioinformatcis and statistical approaches to deal with different situations. The raw reads from sequencing centers need to be processed and denoised by several steps and then be further interpreted by ecological and statistical analysis. So understanding these algorithms and combining different approaches could potentially reduce the influence of noises and biases at different steps. And an efficient and accurate pipeline is important to …
Date:
August 2017
Creator:
Rong, Ruichen
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Evaluating the Role of UV Exposure and Recovery Regimes in PAH Photo-Induced Toxicity to Daphina Magna
Polyaromatic hydrocarbons (PAHs) are contaminants synthesized through incomplete combustion of carbon based substances. PAHs are known to be photodynamic and toxicity increases exponentially when in contact with ultraviolet radiation (UV). The effect of UV absent recovery periods and potential for latent toxicity during photo-induced toxicity are previously unknown and are not included within the toxicity model. Results of equal interval tests further support the current reciprocity model as a good indicator of PAH photo-induced toxicity. Interval test results also indicate a possible presence of time-dependent toxicity and recovery thresholds and should be included into toxicity risk assessments. Moreover, results of latent effects assays show that latent mortality is a significant response to PAH photo-induced toxicity and should be included into toxicity risk assessments. The present research demonstrates that UV exposure time rate is a significant driving force of PAH photo-induced toxicity.
Date:
August 2017
Creator:
Gnau, Jennifer Leigh
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library
Generating Molecular Biology Tools to Investigate the Ca2+ Binding Ability of Arabidopsis TON2
The position of the cell division plane in plants is determined by the position of the preprophase band. The pre prophase band (PPB) is a ring of microtubules centered around the nucleus on the inner side of plasma membrane that establishes the cortical division site. The PPB forms at the end of G2 and breaks down at the end of prophase leaving behind protein markers of its position that are collectively called the cortical division site. During cytokinesis the phragmoplast expands towards the cortical division site and mediates the fusion of the new cell plate with the mother cell at that position. Several proteins necessary for PPB formation in plants have been identified, including maize DCD1 and ADD1 and Arabidopsis TON2, which are all type 2A protein phosphatase (PP2A)B" regulatory subunits. DCD1, ADD1, and TON2 localize to the PPB and the cortical division site through metaphase. The PP2A subunits each have two EF-hand domains, which are predicted to bind calcium ions. Since calcium ions are important for some aspects of cell division, we designed a series of constructs to test if TON2 binds calcium. TON2 protein was cloned into expression vectors, pET42a, and expression of TON2 protein was confirmed via …
Date:
August 2017
Creator:
Shao, Danyang
Object Type:
Thesis or Dissertation
System:
The UNT Digital Library