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  Partner: UNT Libraries
 Department: Department of Biological Sciences
 Decade: 2010-2019
Biodiversity and Genetic Structure of Benthic Macroinvertebrates along an Altitudinal Gradient: A Comparison of the Windhond and Róbalo River Communities on Navarino Island, Chile

Biodiversity and Genetic Structure of Benthic Macroinvertebrates along an Altitudinal Gradient: A Comparison of the Windhond and Róbalo River Communities on Navarino Island, Chile

Date: May 2016
Creator: Pulliam, Lauren
Description: Altitudinal gradients in Sub-Antarctic freshwater systems present unique opportunities to study the effect of distinct environmental gradients on benthic macroinvertebrate community composition and dispersal. This study investigates patterns in biodiversity, dispersal and population genetic structure of benthic macroinvertebrate fauna across an altitudinal gradient between two watersheds on Navarino Island in southern Chile. Patterns in diversity, density, evenness and functional feeding groups were not significantly different across the altitudinal gradient in both the Windhond and Róbalo Rivers. Taxa richness in both rivers generally increased from the headwaters of the river to the mouth, and functional feeding group patterns were consistent with the predictions of the River Continuum Concept. Population genetic structure and gene flow was investigated by sampling the mitochondrial cytochrome oxidase I gene in two invertebrate species with different dispersal strategies. Hyalella simplex (Amphipoda) is an obligate aquatic species, and Meridialaris chiloeense (Ephemeroptera) is an aquatic larvae and a terrestrial winged adult. Contrasting patterns of population genetic structure were observed. Results for Hyalella simplex indicate significant differentiation in genetic structure in the Amphipod populations between watersheds and lower genetic diversity in the Róbalo River samples, which may be a result of instream dispersal barriers. Meridialaris chiloeense exhibited weak population structure ...
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The Effects of Neonicotinoid Exposure on Embryonic Development and Organ Mass in Northern Bobwhite Quail

The Effects of Neonicotinoid Exposure on Embryonic Development and Organ Mass in Northern Bobwhite Quail

Date: May 2016
Creator: Gobeli, Amanda
Description: Since their emergence in the early 1990s, neonicotinoid use has increased exponentially to make them the world's most prevalent insecticides. Although there is considerable research concerning the lethality of neonicotinoids, their sub-lethal and developmental effects are still being explored, especially with regards to non-mammalian species. The goal of this research was to investigate the effects of the neonicotinoid imidacloprid on the morphological and physiological development of northern bobwhite quail (Colinus virginianus). Bobwhite eggs (n = 650) were injected with imidacloprid concentrations of 0 (sham), 10, 50, 100 and 150 grams per kilogram of egg mass, which was administered at day 0 (pre-incubation), 3, 6, 9, or 12 of growth. Embryos were dissected on day 19 when they were weighed, staged, and examined for any overt structural deformities. Embryonic heart, liver, lungs and kidneys were also weighed and preserved for future use. Treated embryos exhibited increased frequency of severely deformed beaks and legs, as well as larger hearts and smaller lungs at the higher dosing concentrations. Some impacts are more pronounced in specific dosing periods, implying that there may be critical windows of development when embryos are highly susceptible to neonicotinoid exposure. This investigation suggests that imidacloprid could play a significant ...
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Genetic Characterization of Central and South American Populations of Scarlet Macaw (Ara macao)

Genetic Characterization of Central and South American Populations of Scarlet Macaw (Ara macao)

Date: May 2016
Creator: Kim, Tracy
Description: The wild populations of the Scarlet Macaw subspecies native to southern Mexico and Central America, A. m. cyanoptera, have been drastically reduced over the last half century and are now a major concern to local governments and conservation groups. Programs to rebuild these local populations using captive bred specimens must be careful to reintroduce the native A. m. cyanoptera, as opposed to the South American nominate subspecies (A. m. macao) or hybrids of the two subspecies. Molecular markers for comparative genomic analyses are needed for definitive differentiation. Here I describe the isolation and sequence analysis of multiple loci from 7 pedigreed A. m. macao and 14 pedigreed A. m. cyanoptera specimens. The loci analyzed include the 18S rDNA genes, the complete mitogenome as well as intronic regions of selected autosomally-encoded genes. Although the multicopy18S gene sequences exhibited 10% polymorphism within all A. macao genomes, no differences were observed between any of the 21 birds whose genomes were studied. In contrast, numerous polymorphic sites were observed throughout the 16,993 bp mitochondrial genomes of both subspecies. Although much of the polymorphism was observed in the genomes of both subspecies, subspecies-specific alleles were observed at a number of mitochondrial loci, including 12S, 16S, ...
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The Influence of Urban Green Spaces on Declining Bumble Bees (Hymenoptera: Apidae)

The Influence of Urban Green Spaces on Declining Bumble Bees (Hymenoptera: Apidae)

Date: May 2016
Creator: Beckham, Jessica Lorene
Description: Bumble bees (Bombus spp.) are adept pollinators of countless cultivated and wild flowering plants, but many species have experienced declines in recent decades. Though urban sprawl has been implicated as a driving force of such losses, urban green spaces hold the potential to serve as habitat islands for bumble bees. As human populations continue to grow and metropolitan areas become larger, the survival of many bumble bee species will hinge on the identification and implementation of appropriate conservation measures at regional and finer scales. North Texas is home to some the fastest-growing urban areas in the country, including Denton County, as well as at least two declining bumble bee species (B. pensylvanicus and B. fraternus). Using a combination of field , molevular DNA and GIS methods I evaluated the persistence of historic bumble bee species in Denton County, and investigated the genetic structure and connectivity of the populations in these spaces. Field sampling resulted in the discovery of both B. pensylvanicus and B. fraternus in Denton County's urban green spaces. While the relative abundance of B. fraternus in these spaces was significantly lower than historic levels gleaned from museum recors, that of B. pensylvanicus was significantly higher. Statistical analyses found ...
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Informing Conservation Management Using Genetic Approaches: Greater Sage-grouse and Galápagos Short-eared Owls as Case Studies

Informing Conservation Management Using Genetic Approaches: Greater Sage-grouse and Galápagos Short-eared Owls as Case Studies

Date: May 2016
Creator: Schulwitz, Sarah E
Description: Small isolated populations are of particular conservation interest due to their increased extinction risk. This dissertation investigates two small wild bird populations using genetic approaches to inform their conservation. Specifically, one case study investigated a Greater Sage-grouse (Centrocercus urophasianus) population located in northwest Wyoming near Jackson Hole and Grand Teton National Park. Microsatellite data showed that the Jackson sage-grouse population possessed significantly reduced levels of neutral genetic diversity and was isolated from other Wyoming populations. Analysis with single nucleotide polymorphisms (SNPs) and microsatellite data provided further evidence that the population's timing of isolation was relatively recent and most likely due to recent anthropogenic habitat changes. Conservation recommendations include maintaining or increasing the population's current size and reestablishing gene flow with the nearest large population. The second case study investigated the genetic distinctiveness of the Floreana island population of the Galápagos Short-eared Owl (Asio flammeus galapagoensis). Mitochondrial DNA sequence data did not detect differences across nine island populations, yet microsatellite and morphometric data indicated that limited gene flow existed with the population and surrounding island populations, which appeared asymmetric in direction from Floreana to Santa Cruz with no indication of gene flow into Floreana. These results have important conservation implications and ...
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The Effect of Post-exercise Ethanol Consumption on the Acute Hormonal Response to Heavy Resistance Exercise in Women

The Effect of Post-exercise Ethanol Consumption on the Acute Hormonal Response to Heavy Resistance Exercise in Women

Date: December 2015
Creator: Budnar, Jr., Ronald G.
Description: The purpose of this study was to examine the hormonal response to acute ethanol ingestion following a bout of heavy resistance exercise in women. Eight resistance trained women completed two identical acute heavy resistance exercise tasks (AHRET). From 10-20 minutes post-AHRET, participants consumed either a grain ethanol or a placebo beverage. Blood was collected before (PRE) and immediately after the AHRET (IP) and then every 20 minutes for five hours. Blood collected after beverage ingestion was pooled into 3 batches (phases: 20-40 minutes, 60-120 minutes, and 140-300 minutes post-exercise) and analyzed for serum total testosterone (TT), free testosterone (FT), insulin-like growth factor-I (IGF-I), human growth hormone (GH), cortisol (COR), and estradiol (E2) concentrations. Circulating concentrations of TT were significantly greater at P20-40 than at PRE, P60-120, and P140-300. Circulating concentrations of FT were significantly greater at P20-40 than at all other times. Circulating concentrations of GH were significantly greater at IP than at PRE, P60-120, and P140-300. Circulating concentrations of COR were significantly greater at P20-40 than at all other times. Additionally, COR concentrations at P140-300 were significantly lower than at all other times. Circulating concentrations of IGF-1 were significantly greater at P20-40 than at P60-120 and P140-300. Circulating concentrations ...
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Environmental Modulation of the Onset of Air-breathing of the Siamese Fighting Fish and the Blue Gourami

Environmental Modulation of the Onset of Air-breathing of the Siamese Fighting Fish and the Blue Gourami

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Date: December 2015
Creator: Mendez Sanchez, Jose Fernando
Description: This study determined the effect of hypoxia on air-breathing onset and physiological and morphological characters in larvae of the air breathing fishes Trichopodus trichopterus and Betta splendens. Larvae were exposed intermittently (12/12 h daily) to 20, 17, and 14 kPa of PO2 from 1 to 40 days post-fertilization. Survival, onset of air breathing, wet body mass, O2, Pcrit were measured every 5 dpf. Hypoxia advanced by 4 days, and delayed by 9 days, the onset of air breathing in Betta and Trichopodus, respectively. Hypoxia increased larval body length, wet mass, and labyrinth organ respiratory surface of Betta, but did not affect these factors in Trichopodus. Hypoxic exposure increased O2 by 50-100% at each day throughout larval development in Betta, but had no effect on larval Trichopodus. Hypoxia decreased Pcrit in Betta by 37%, but increased Pcrit in Trichopodus by 70%. Larval Betta reared in hypoxia showed a modified heart rate:opercular rate ratio (3:1 to 2:1), but these changes did not occur in Trichopodus. Compared to Betta, the blood of Trichopodus had a higher P50 and much smaller Bohr and Root effects. These interspecific differences are likely due to ecophysiological differences: Betta is a non- obligatory air-breather after 36 dpf with ...
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Identification and Characterization of an Arabidopsis Thaliana Mutant with Tolerance to N-lauroylethanolamime

Identification and Characterization of an Arabidopsis Thaliana Mutant with Tolerance to N-lauroylethanolamime

Date: December 2015
Creator: Adhikari, Bikash
Description: N-Acylethanolamines (NAEs) are fatty acid derivatives in plants that negatively influence seedling growth. N-Lauroylethanolamine (NAE 12:0), one type of NAE, inhibits root length, increases radial swelling of root tips and reduces root hair numbers in a dose dependent manner in Arabidopis thaliana L. (ecotype Columbia). A forward genetics approach was employed by screening a population of T-DNA “activation-tagged” developed by the Salk Institute lines for NAE resistance to identify potential genes involved in NAE signaling events in Arabidopsis thaliana L. (ecotype Columbia). Seeds of the activation tagged lines were grown at 0, 25, 30, 50, 75 and 100 µM N-lauroylethanolamime (NAE 12:0). Ten plants which displayed NAE tolerance (NRA) seedling phenotypes, compared with wildtype (Columbia, Col-0) seedlings were identified. I focused on one mutant line, identified as NRA 25, where the tolerance to NAE 12:0 appears to be mediated by a single dominant, nuclear gene. Thermal asymmetric interlaced (TAIL) PCR identified the location of the T-DNA insert as 3.86 kbp upstream of the locus At1g68510. Quantitative PCR indicated that the transcript level corresponding to At1g68510 is upregulated approximately 20 fold in the mutant relative to wildtype. To determine whether the NAE tolerance in NRA 25 is associated with overexpression of ...
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Identification of Genes Involved in Flocculation by Whole Genome Sequencing of Thauera Aminoaromatica Strain Mz1t Floc-defective Mutants

Identification of Genes Involved in Flocculation by Whole Genome Sequencing of Thauera Aminoaromatica Strain Mz1t Floc-defective Mutants

Date: December 2015
Creator: Prombutara, Pinidphon
Description: Thauera aminoaromatica MZ1T, a floc-forming bacterium isolated from an industrial activated sludge wastewater treatment plant, overproduces exopolysaccharide (EPS) leading to viscous bulking. This phenomenon results in poor sludge settling and dewatering during the clarification process. To identify genes responsible for bacterial flocculation, a whole genome phenotypic sequencing technique was applied. Genomic DNA of MZ1T flocculation-deficient mutants were subjected to massively parallel sequencing. The resultant high-quality reads were assembled and compared to the reference genome of the wild type genome. We identified nine nonsynonymous mutations and one nonsense mutation putatively involved in EPS biosynthesis. Complementation of the nonsense mutation located in an EPS deacetylase gene restored the flocculating phenotype. The FTIR spectra of EPS isolated from the wild-type showed reduced C=O peak of the N-acetyl group at 1665 cm-1 as compared to the spectra of MZ1T floc-deficient mutant EPS, suggesting that the WT EPS was partially deacetylated. Gene expression analysis also demonstrated the deacetylase gene transcript increased before flocculation occurred. The results suggest that the deacetylation of MZ1T EPS is crucial for flocculation. The information obtained from this study will be useful for preventing viscous bulking and wastewater treatment system failure, and may have potential applications in the biotechnology sector for ...
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Identification of Hox Genes Controlling Thrombopoiesis in Zebrafish

Identification of Hox Genes Controlling Thrombopoiesis in Zebrafish

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Date: December 2015
Creator: Sundaramoorthi, Hemalatha
Description: Thrombocytes are functional equivalents of mammalian platelets and also possess megakaryocyte features. It has been shown earlier that hox genes play a role in megakaryocyte development. Our earlier microarray analysis showed five hox genes, hoxa10b, hoxb2a, hoxc5a, hoxc11b and hoxd3a, were upregulated in zebrafish thrombocytes. However, there is no comprehensive study of genome wide scan of all the hox genes playing a role in megakaryopoiesis. I first measured the expression levels of each of these hox genes in young and mature thrombocytes and observed that all the above hox genes except hoxc11b were expressed equally in both populations of thrombocytes. hoxc11b was expressed only in young thrombocytes and not in mature thrombocytes. The goals of my study were to comprehensively knockdown hox genes and identify the specific hox genes involved in the development of thrombocytes in zebrafish. However, the existing vivo-morpholino knockdown technology was not capable of performing such genome-wide knockdowns. Therefore, I developed a novel cost- effective knockdown method by designing an antisense oligonucleotides against the target mRNA and piggybacking with standard control morpholino to silence the gene of interest. Also, to perform knockdowns of the hox genes and test for the number of thrombocytes, the available techniques were ...
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The Role of Cysteinyl Leukotriene Receptor 2 in Thrombocyte Aggregation

The Role of Cysteinyl Leukotriene Receptor 2 in Thrombocyte Aggregation

Date: December 2015
Creator: Reyna, Julianna
Description: Cysteinyl leukotriene receptor 2, a G-protein coupled receptor known to be expressed and functional on human platelets. However, it seems that upon ligand activation the cysteinyl leukotriene receptor 2 activates a variety of signaling pathways in multiple cell types among different species. Previously, a former laboratory member Vrinda Kulkarni found cysteinyl leukotriene receptor 2 to be expressed on the surface of adult zebrafish thrombocytes. In this work I studied the characteristics of aggregation in adult zebrafish thrombocytes with the knockdown of cysteinyl leukotriene receptor 2. I used a newly developed knockdown method to study the function of cysteinyl leukotriene receptor 2. Knockdown of the cysteinyl leukotriene was confirmed using RT-PCR results showed p=.001, reduced sell surface level of expression of the cysteinyl leukotriene receptor 2 results showed that p=.002. I found that the knockdown of cysteinyl leukotriene receptor 2 results in prothrombotic thrombocytes by using flow cytometry p=.0001.
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Role of Gpr17 in Thrombocyte Aggregation in Adult Zebrafish

Role of Gpr17 in Thrombocyte Aggregation in Adult Zebrafish

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Date: December 2015
Creator: Bohassan, Maruah Hejey
Description: GPR17, a uracil nucleotide cysteinyl leukotriene receptor, belongs to the GPCR (G protein coupled receptor) family. It has been shown recently that inhibiting this protein in the nervous system in mice can lead to blockage of oligodendrocyte maturation, which supports myelin repair. Interestingly, our laboratory found GPR17 in thrombocytes. However, we do not know whether it has any function in thrombocyte aggregation or the nature of the ligand. In this paper, we studied the role of GPR17 in hemostasis, which is a fundamental defense mechanism in the event of injury. Using zebrafish as a model system, our laboratory has studied specifically thrombocytes, which play a significant role in hemostasis. The major reasons to use zebrafish as a model system are that their thrombocytes are functionally equivalent to human platelets, the adult fish are amenable to knockdown experiments, and they are readily available in the market. This study was performed by using a piggy back knockdown method where we used a chemical hybrid of control morpholino and an antisense oligonucleotide sequence leads to the degradation the mRNA for GPR17. After knockdown GPR17 in thrombocytes, the percent difference of the thrombocytes aggregation between the control and knockdown blood samples was measured by ...
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Thresholds and Legacy Effects of Tropical Floodplain Fish Assemblages in Response to Flood Attributes

Thresholds and Legacy Effects of Tropical Floodplain Fish Assemblages in Response to Flood Attributes

Date: December 2015
Creator: Hoeinghaus, Ana Paula Ferrari
Description: Natural flow regimes are critical for sustaining biodiversity and river integrity. Floods and droughts form an important component of river systems and control population sizes and species diversity across space and time. Modification of flow regimes, including disruption of the timing, magnitude and duration of flooding, is a global problem, and many new impoundments are planned for large river-floodplain ecosystems in the tropics. Flow modifications may cause dramatic non-linear responses in population sizes and have lasting effects through time, but such topics are poorly investigated over multi-year scales, especially in highly diverse tropical ecosystems. Using a long-term dataset from the Upper Paraná River floodplain, Brazil, I tested for threshold and legacy effects of fish assemblages to flood attributes, such as timing, magnitude, duration, rate of change and variation. Specifically, I hypothesized that long duration, high magnitude floods would elicit threshold responses in long-distance migratory fish species and these responses result in significant legacy effects detectable over multiple years. Consistent positive threshold responses to increasing flood duration and magnitude were detected for many species and not significantly correlated with reproductive guilds. Legacy effects were prevalent (i.e. identified for more than 90% of species) and including flood attributes from previous years increased ...
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Dynamics of Stream Fish Metacommunities in Response to Drought and Reconnectivity

Dynamics of Stream Fish Metacommunities in Response to Drought and Reconnectivity

Date: August 2015
Creator: Driver, Lucas J.
Description: This dissertation investigates the spatio-temporal dynamics of intermittent stream fish metacommunities in response drought-induced fragmentation and re-connectivity using both field and experimental approaches. A detailed field study was conducted in two streams and included pre-drought, drought, and post-drought hydrological periods. Fish assemblages and metacommunity structure responded strongly to changes in hydrological conditions with dramatic declines in species richness and abundance during prolonged drought. Return of stream flows resulted in a trend toward recovery but ultimately assemblages failed to fully recover. Differential mortality, dispersal, recruitment among species indicates species specific responses to hydrologic fragmentation, connectivity, and habitat refugia. Two manipulative experiments tested the effects of drought conditions on realistic fish assemblages. Fishes responded strongly to drought conditions in which deeper pools acted as refugia, harboring greater numbers of fish. Variability in assemblage structure and movement patterns among stream pools indicated species specific habitat preferences in response predation, resource competition, and desiccation. Connecting stream flows mediated the impacts of drought conditions and metacommunity dynamics in both experiments. Results from field and experimental studies indicate that stream fish metacommunities are influenced by changes in hydrological conditions and that the timing, duration, and magnitude of drought-induced fragmentation and reconnecting stream flows have important consequences ...
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Effect of Acute Alcohol Ingestion on Resistance Exercise Induced Mtorc1 Signaling in Human Muscle

Effect of Acute Alcohol Ingestion on Resistance Exercise Induced Mtorc1 Signaling in Human Muscle

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Date: August 2015
Creator: Anthony A. Duplanty
Description: The purpose of this project was to further elucidate the effects post-exercise alcohol ingestion. This project had many novel aspects including using a resistance exercise (RE) only exercise design and the inclusion of women. To our knowledge, we are the first to investigate the effect of post-RE alcohol ingestion in women. In the first chapter of this project, information on the prevalence of alcohol use and the importance of skeletal muscle as a dynamic and metabolic tissue was provided. In chapter two, the effects of post-RE alcohol ingestion in men and women are detailed. The major findings of this study was that although RE elicited similar mTORC1 signaling both in men and in women, alcohol ingestion appeared to only attenuate RE-induced phosphorylation of the mTORC1 signaling pathway in men. The third chapter focused on examining the effects of post-RE alcohol ingestion on acute testosterone bioavailability. The primary findings of this study was that alcohol substantially elevated serum total and free testosterone concentrations during recovery from a bout of resistance exercise. The fourth chapter detailed factors that contribute to bone density in men. The major findings of this study was that young adult male long-distance runners who participated in resistance training ...
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Glucose Induces Sensitivity to Oxygen Deprivation and Alters Gene Expression in Caenorhabditis Elegans

Glucose Induces Sensitivity to Oxygen Deprivation and Alters Gene Expression in Caenorhabditis Elegans

Date: August 2015
Creator: Garcia, Anastacia M.
Description: An organisms’ diet represents an exogenous influence that often yields colossal effects on long-term health and disease risk. The overconsumption of dietary sugars for example, has contributed to significant increases in obesity and type-2 diabetes; health issues that are costly both economically and in terms of human life. Individuals who are obese or are type-2 diabetic often have compromised oxygen delivery and an increased vulnerability to oxygen-deprivation related complications, such as ischemic strokes, peripheral arterial disease and myocardial infarction. Thus, it is of interest to identify the molecular changes glucose supplementation or hyperglycemia can induce, which ultimately compromise oxygen deprivation responses. By utilizing the Caenorhabditis elegans genetic model system, which is anoxia tolerant, I determined that a glucose-supplemented diet negatively impacts responses to anoxia and that the insulin-like signaling pathway, through fatty acid and ceramide biosynthesis and antioxidant activity, modulates anoxia survival. Additionally, a glucose-supplemented diet induces lipid accumulation. Use of RNA-sequencing analysis to compare gene expression responses in animals fed either a standard or glucose-supplemented diet revealed that glucose impacts the expression of genes involved with multiple cellular processes including lipid and carbohydrate metabolism, stress responses, cell division, and extracellular functions. Several of the genes we identified are homologous ...
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Metabolism and Action of Polyunsaturated N-acylethanolamines in Arabidopsis Thaliana Seedlings

Metabolism and Action of Polyunsaturated N-acylethanolamines in Arabidopsis Thaliana Seedlings

Date: August 2015
Creator: Keereetaweep, Jantana
Description: The lipoxygenase (LOX) pathway plays an important role in the oxidative metabolism of polyunsaturated N-acylethanolamines (PU-NAEs). The LOX pathway functions in conjugation with hydrolysis by fatty acid amide hydrolase (FAAH) and to produce oxidized NAEs during seed germination and early seedling development. When Arabidopsis seedlings were grown in low micromolar concentrations of lauroylethanolamide (NAE 12:0), growth retardation and elevated endogenous PU-NAE levels were observed due to the competitive inhibition of LOX by NAE 12:0. The elevated levels of endogenous PU-NAEs were more pronounced in genotypes with reduced NAE hydrolase capacity (faah knockouts), and less evident with overexpression of FAAH. Alterations in PU-NAE metabolism were studied in seedlings of various lox and FAAH mutants. The partitioning of PU-NAEs into oxylipin metabolites was exaggerated in the presence of exogenous linolenoylethanolamide (NAE18:3) and resulted in bleaching of cotyledons. The bleaching phenotype was restricted to a narrow developmental window (3-to-5 days after sowing), and was attributed to a reversible disruption of thylakoid membranes in chloroplasts. Biochemical and genetic evidence suggested that 9-hydro(pero)xy and 13-hydro(pero)xy octadecatrienoylethanolamides (9- and 13-NAE-H(P)OT), but not their corresponding hydro(pero)xy free fatty acids, induced cotyledon bleaching. The LOX-mediated metabolites of NAE18:3 shared some overlapping effects on seedling development with those of ...
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Characteristics of Primary Cilia and Centrosomes in Neuronal and Glial Lineages of the Adult Brain

Characteristics of Primary Cilia and Centrosomes in Neuronal and Glial Lineages of the Adult Brain

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Date: May 2015
Creator: Bhattarai, Samip Ram
Description: Primary cilia are sensory organelles that are important for initiating cell division in the brain, especially through sonic hedgehog (Shh) signaling. Several lines of evidence suggest that the mitogenic effect of Shh requires primary cilia. Proliferation initiated by Shh signaling plays key roles in brain development, in neurogenesis in the adult hippocampus, and in the generation of glial cells in response to cortical injury. In spite of the likely involvement of cilia in these events, little is known about their characteristics. Centrosomes, which are associated with primary cilia, also have multiple influences on the cell cycle, and they are important in assembling microtubules for the maintenance of the cell’s cytoskeleton and cilia. The cilia of terminally differentiated neurons have been previously examined with respect to length, incidence, and receptors present. However, almost nothing is known about primary cilia in stem cells, progenitors, or differentiated glial cells. Moreover, it is not known how the properties of cilia and centrosomes may vary with cell cycle or proliferative potential, in brain or other tissues. This dissertation focuses first on neurogenesis in the hippocampal subgranular zone (SGZ). The SGZ is one of the few brain regions in mammals that gives rise to a substantial ...
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Cytotoxicity and Functional Toxicity of Mefloquine and Search for Protective Compounds

Cytotoxicity and Functional Toxicity of Mefloquine and Search for Protective Compounds

Date: May 2015
Creator: Holmes, Katelyn
Description: Mefloquine hydrochloride is an antimalarial agent that has been used for the past 40 years. Numerous reports of neurological side effects have recently led the FDA to issue a strong warning regarding long-term neurological effects. This warning lead to the U.S. Army’s Special Forces and other components to discontinue its use in July of 2013. Despite reported adverse side effects, mefloquine remains in circulation and is recommended to travelers going to specific Asian countries. Mefloquine has been used as a treatment for those already infected with the malaria parasite (blood concentrations ranging from 2.1 to 23 µM), and as prophylaxis (blood concentrations averaging 3.8 µM) (Dow 2003). The purpose of this study was to quantify Mefloquine’s toxicity using spontaneously active nerve cell networks growing on microelectrode arrays in vitro and to identify compounds that alleviate or reduce toxic effects. The current literature on mefloquine toxicity is lacking electrophysiological data. These data will contribute to research on the mechanism of adverse side effects associated with mefloquine use. Sequential titration experiments were performed by adding increasing concentrations of mefloquine solution to cultured neurons. Network responses were quantified and reversibility was examined. In each network, activity decreases were normalized as a percent of ...
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The Effect of Curcumin Supplementation on Physical and Biological Indices of Delayed Onset Muscle Soreness and Inflammation Following Muscle Injury

The Effect of Curcumin Supplementation on Physical and Biological Indices of Delayed Onset Muscle Soreness and Inflammation Following Muscle Injury

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Date: May 2015
Creator: Venable, Adam Steven
Description: In this project, the effects of dietary polyphenols on exercise-induced muscle damage and vascular health are examined. Dietary polyphenols exert well-known anti-inflammatory effects; however, how these effects are realized with respect to vascular health and EIMD is relatively unknown. I begin by reviewing the available literature surrounding the impact of three dietary polyphenols (curcumin, catechins, and quercetin) on inflammation associated with EIMD. It is well established that their primary means of anti-inflammation is through alterations of NF-κB and AP-1 transcription activities. Given this, their inclusion into training strategies seems reasonable. Consistent evidence is presented making a case for the anti-inflammatory effects of dietary polyphenols following EIMD. I follow this review up by completing an in-depth study on the consumption of curcumin prior to EIMD. I found curcumin (1000 mg/day) can reduce subjective soreness and decrease inflammation compared to placebo controls. To further understand the effects of dietary polyphenols on health, I investigate the effects of a four-week supplementation period of cocoa (catechins) on vascular. I concluded that atherogenic risk in obese women is reduced after consumption of cocoa. In addition to these experimental projects, I developed two novel methods that can be used to investigate vascular health (EMP concentration) and ...
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Improving Processing Efficiency for Forensic DNA Samples

Improving Processing Efficiency for Forensic DNA Samples

Date: May 2015
Creator: Connon, Catherine Cupples
Description: The goal of this project was to reduce processing time for forensic DNA testing without incurring significant added costs and/or the need for new instrumentation, while still generating high quality profiles. This was accomplished by: 1) extraction normalization using the ChargeSwitch® Forensic DNA Purification Kit such that a small range of DNA concentrations was consistently obtained, eliminating the need for sample quantification and dilution; 2) developing fast PCR protocols for STR primer sets using shorter amplification methods, low volume reactions and non-fast thermal cyclers; and 3) developing a quicker 3130xl Genetic Analyzer detection method using an alternative polymer/array length combination. Extraction normalization was achieved through a reduction in bead quantity, thereby forcing an increase in bead binding efficiency. Four products (AmpliTaq Gold® Fast PCR Master Mix, KAPA2G™ Fast Multiplex PCR Kit, SpeedSTAR™ HS DNA Polymerase and Type-it Microsatellite PCR Kit) were evaluated for low volume (3μl) fast PCR on a 384-well Veriti® thermal cycler with the Identifiler primer set. KAPA2G™ was selected for 3μl fast PCR protocols using PowerPlex 16 HS and Identifiler Plus primer sets (42-51min), as well as 5μl and 6μl Identifiler fast reactions on a 9700 thermal cycler (51-60min). Alternative detection (POP-6™/22cm) achieved 24-28min run times, but ...
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Manipulations of Sucrose/proton Symporters and Proton-pumping Pyrophosphatase Lead to Enhanced Phloem Transport But Have Contrasting Effects on Plant Biomass

Manipulations of Sucrose/proton Symporters and Proton-pumping Pyrophosphatase Lead to Enhanced Phloem Transport But Have Contrasting Effects on Plant Biomass

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Date: May 2015
Creator: Khadilkar, Aswad S
Description: Delivery of photoassimilate, mainly sucrose (Suc) from photoautotrophic source leaves provides the substrate for the growth and maintenance of sink tissues such as roots, storage tissues, flowers and fruits, juvenile organs, and seeds. Phloem loading is the energized process of accumulating solute in the sieve element/companion cell complex of source leaf phloem to generate the hydrostatic pressure that drives long-distance transport. In many plants this is catalyzed by Suc/Proton (H+) symporters (SUTs) which are energized by the proton motive force (PMF). Overexpression of SUTs was tested as means to enhance phloem transport and plant productivity. Phloem specific overexpression of AtSUC2 in wild type (WT) tobacco resulted in enhanced Suc loading and transport, but against the hypothesis, plants were stunted and accumulated carbohydrates in the leaves, possibly due to lack of sufficient energy to support enhanced phloem transport. The energy for SUT mediated phloem loading is provided from the PMF, which is ultimately supplied by the oxidation of a small proportion of the loaded photoassimilates. It was previously shown that inorganic pyrophosphate (PPi) is necessary for this oxidation and overexpressing a proton-pumping pyrophosphatase (AVP1) enhanced both shoot and root growth, and augmented several energized processes like nutrient acquisition and stress responses. ...
Contributing Partner: UNT Libraries
The Microbial Retting Environment of Hibiscus Cannabinus and Its Implications in Broader Applications

The Microbial Retting Environment of Hibiscus Cannabinus and Its Implications in Broader Applications

Date: May 2015
Creator: Visi, David K.
Description: Fiber-yielding plants is an area of increased interest due to the potential use in a variety of green-based materials. These biocomposites can be incorporated into multiple uses; for example, to replace building materials and interior vehicular paneling. The research here aims to focus in on the crop Hibiscus cannabinus for utilization into these functions. H. cannabinus is economically attractive due to the entire process being able to be accomplished here in the United States. The plant can be grown in a relatively short growth period (120-180 days), and then processed and incorporated in a biocomposite. The plant fiber must first be broken down into a useable medium. This is accomplished by the retting process, which occurs when microbial constituents breakdown the heteropolysaccharides releasing the fiber. The research aims to bridge the gap between the primitive process of retting and current techniques in molecular and microbiology. Utilizing a classical microbiological approach, which entailed enrichment and isolation of pectinase-producing bacteria for downstream use in augmented microbial retting experiments. The tracking of the bacteria was accomplished by using the 16S rRNA which acts as “barcodes” for bacteria. Next-generation sequencing can then provide data from each environment telling the composition and microbial diversity of ...
Contributing Partner: UNT Libraries
Neurotoxicity of the Industrial Solvent 4-Methylcyclohexanemethanol: Involvement of the GABA Receptor

Neurotoxicity of the Industrial Solvent 4-Methylcyclohexanemethanol: Involvement of the GABA Receptor

Date: May 2015
Creator: Gibson, Jason
Description: A recent chemical spill of 4-Methylcyclohexanemethanol (4-MCHM) in West Virginia left 300,000 people without water. Officials claimed that this compound is not lethally toxic, but potentially harmful if swallowed or inhaled, and can cause eye and skin irritation. Sittig's Handbook of Toxic and Hazardous Chemical Carcinogens reports high exposures from skin contact or inhalation may cause damage to the heart, liver, kidneys, and lungs, and may result in death. However, no quantitative data seem to exist and no references can be found on neurotoxicity. We have investigated the neurotoxicity of 4-MCHM using mammalian nerve cell networks grown on microelectrode arrays. Network spontaneous activity from multiple units (range 48 – 120 per network) were used as the primary readout. Individual units were followed based on spike waveforms digitized at 40 kHz (Plexon MNAP system). Dose response curves show the effective inhibitory concentration at 50 percent decrease (EC50) to average 27.4 microM SD±6.17. However, in the presence of 40 microM bicuculline, a competitive GABAA antagonist, the EC50 shifts to 70.63uM SD ±4.3; implying that early, low concentration exposures to 4-MCHM involve GABA activation. Initial activity loss occurs without active unit loss (defined as 10 or more template threshold crossing per min), indicating ...
Contributing Partner: UNT Libraries
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