Oxygen availability is critical for survival for most organisms. The nematode, C. elegans, has been useful for studying genetic regulation of anoxia tolerance due to the oxygen deprivation response mechanisms shared with other metazoans. Studies examining long-term anoxia (72h, LTA) tolerance have only been conducted at adult day 1. To investigate the effect of aging on anoxia tolerance wild-type and mutant strains were exposed to LTA between adult day 1 and day 9. Wild-type isolates and daf-16(mu86) (FOXO transcription factor regulated by insulin-signaling) and aak-2(gt33) (catalytic subunit of AMP-activated protein kinase) strains were anoxia sensitive at day 1 and displayed increased LTA tolerance with aging correlated with reproductive senescence followed by a decline in survivorhsip through day 9. The daf-2(e1370) (insulin receptor homologue of C. elegans), glp-1(e2141) (a lin-12/Notch receptor) and fog-2(q71) (required for spermatogenesis) strains were LTA-tolerant through day 5. I conclude that aging influences LTA-tolerance in a strain- and age-dependent manner. In addition to being LTA-tolerant the daf-2(e1370) and glp-1(e2141) strains have a longevity phenotype that is suppressed by loss of kri-1 or daf-12. While loss of kri-1 did not suppress the LTA-tolerant phenotype of glp-1(e2141) at day 1 the portion of impaired survivors increased at day 3 …

The northern bobwhite (Colinus virginianus) population decline is a severe, rangewide phenomenon beginning >150 years ago and continuing today. In this investigation, I: 1. document the timeline of bobwhite population decline and unintended genetic consequences of attempted remedies, 2) develop a model useful for predicting possible locations of potentially sustainable bobwhite populations in semiarid rangeland in Texas and Oklahoma, and 3) examine the relationship between population monitoring data and meteorological factors. While breeding season call counts of male bobwhite have been used for >70 years to provide estimates of fall populations for hunting, most studies of call counts have focused on mathematics and statistical accuracy of the count, largely overlooking the influence of meteorological factors on call counts. Here, I present the results of >4,400 individual point counts and examine their relationship with meteorological variables recorded at each stop. Humidity was positively correlated with the number of birds recorded (ρ = 0.275, p < 0.001) and temperature was negatively correlated (ρ = -0.252, p < 0.001). The number of birds recorded was significantly higher in wet years than in drought years. There was no significant correlation between wind velocity and number of birds recorded. These results suggest that, while weather …

Factors currently at the forefront of human in vitro fertilization (IVF) that collectively influence treatment success in the form of blastocysts development were investigated during early mammalian embryology with concentration on infertile patients presenting with diminished ovarian reserve or preliminary ovarian failure. A novel experimental technique, Graft Transplant-Embryonic Stem Cells (GT-ESC) was introduced in the mouse model, as the first inclusive approach for embryo selection in IVF treatments resulting in successful graft integration of sibling cells, stage-dependent (day 4) blastocysts. E-Cadherin-catenin bonds play an integral role in trophectoderm cell viability and calcium removal, inducing disruption of cell-to-cell bonds at the blastocyst stage was detrimental to continued blastocyst development. One of the leading methods for embryo selection for uterine transfer in human IVF is application of pre-implantation genetic screening (PGS) methods such as next generation sequencing (NGS). Female patients <35 y do not benefit from this treatment when outcome is measured by presence of fetal heart beats at 10 weeks of gestation. Patients 35-37 y benefit from PGS with no significant difference of outcome based on form of PGS method utilized. Therefore, small nucleotide polymorphism array (snp-array) or targeted-NGS should be selected for this age range to lessen the financial burden …

Postmortem interval (PMI) estimation is a necessary but often difficult task that must completed during a death investigation. The level of difficulty rises as time since death increases, especially with the case of skeletonized remains (long PMI). While challenging, a reliable PMI estimate may be of great importance for investigative direction and cost-savings (e.g. suspect identification, tailoring missing persons searches, non-forensic remains exclusion). Long PMI can be estimated by assessing changes in the organic content of bone (i.e. collagen), which degrades and is lost as the PMI lengthens. Visible-near infrared (VNIR) spectroscopy is one method that can be used for analyzing organic constituents, including proteins, in solid specimens. A 2013 preliminary investigation using a limited number of human cortical bone samples suggested that VNIR spectroscopy could provide a fast, reliable technique for assessing PMI in human skeletal remains. Clear separation was noted between "forensic" and "archaeological" specimen spectra within the near-infrared (NIR) bands. The goal of this research was to develop reliable multivariate classification models that could assign skeletal remains to appropriate PMI classes (e.g. "forensic" and "non-forensic"), based on NIR spectra collected from human cortical bone. Working with a large set of cortical samples (n=341), absorbance spectra were collected …

The anthropogenic alteration of riverine ecosystems has led to declines in the abundance and diversity of freshwater mussels (Bivalvia: Unionoida) worldwide. Central Texas is home to a diverse freshwater mussel fauna including three candidates for federal listing under the Endangered Species Act. Surveys conducted over the last few decades suggest many of the endemic freshwater mussel species in Texas exist in small isolated populations that may be vulnerable to the deleterious effects of genetic diversity loss. Microsatellite primers from two closely related species were used to identify a set of genetic markers that functioned in the Golden Orb (Quadrula aurea). Microsatellite markers were then applied to document the population genetic structure of Q. aurea within and among three connected river drainages in southeastern Texas. Gene flow within existing Q. aurea populations appears high indicating little potential for genetic issues stemming from isolation and inbreeding. Two weakly divergent admixed populations were identified occupying the San Antonio and Guadalupe/San Marcos rivers. Population genetic structure was related to river basin affiliation, but results for environmental factors were unresolved. Current effective population size estimates are large for the Guadalupe/San Marcos drainage and moderately large for the San Antonio drainage and there is no clear …

Cardiovascular activities may increase the brain blood flow improving neuronal activities leading to improved cognition. Consequently, the effects of an acute bout of moderate intensity aerobic exercise on brain hemodynamics and its correlation with cognitive color-word Stroop task performance were tested. The Stroop tasks were congruent (color matches word) and incongruent (color does not match word). Prefrontal (PFC) and motor cortex (MC) blood flow was recorded by fNIRS (functional near-infrared spectroscopy) while the subject was performing the Stroop tasks before and after the 30 minutes of exercise or equivalent time of rest controls (checking for practice effects). Ninety human subjects of age 24± 6, 20 ADHD (attention-deficit hyper-activity disorder), 27 High-BMI (>25), 29 males were recruited. Reaction time ‘RT' decreased (p<0.05) after exercise for both the congruent (12%) and incongruent (10%) Stroop tasks, compared to 8% with practice alone. Accuracy did not change after practice or exercise. HR changes after exercise correlated (p<0.05) with better accuracy and faster RT for the incongruent Stroop task. In general, a metabolic lag occurred in the neuronal deoxy- hemoglobin (Hb) signals behind the systemic oxy-Hb signals. PFC showed the highest effect sizes of Stroop task-responsive systemic hemodynamic changes compared to baseline irrespective of rest …

Through the use of natural history records, published literature, and personal sampling (2011-2016) a total of 454 caddisfly species represented by 24 families and 93 genera were documented from the south-central United States. Two Hydroptilidae species were collected during the 2011-2016 collection efforts that are new to the region: Hydroptilia scheringi and Mayatrichia tuscaloosa. Eightteen species are endemic and 30 are considered species of concern by either federal or state agencies. The majority of each of these groups is Hydroptilidae, or microcaddisflies. Trichoptera community structure, by minimum number of species, was analysed in conjunction with large-scale geographical factors to determine which factor illustrated caddisfly community structure across the region. Physiographic provinces compared to other geographic factors analyzed best-represented caddisfly communities with a minimum of 10 or more species. Statistically, Hydrologic Unit Code 4 (HUC 4) was the most significant geographical factor but low number of samples representing this variable rendered it less representative of caddisfly community structure for the study area.

The purpose of my research work was to assess a variety of Northern Thailand plants tissue extracts for antibacterial and anti-platelet aggregation activity. The Minimum Inhibitory Concentration assay method was used to assess antimicrobial activity of plant extracts, while the Zebrafish Platelet Aggregation Assay and the in vitro Whole Human Blood Impedance Aggregation Assay were used to study anti-platelet activity. Forty one plant extracts harvested from the tissues of 26 plants collected from Northern Thailand were assessed. Thirty-four plant extracts were found to have antibacterial activity against the Gram positive bacteria Staphylococcus aureus and/or Bacillus subtilis, while six plant extracts demonstrated activity against the Gram negative bacterium, Escherichia coli. Thirteen plant extracts exhibited anti-platelet aggregation activity better than the positive control. Two crude plant extracts, twigs from Garcinia sp. and twigs from Goniothalamus chilensis were selected for fractionation. Five of the 12 fractions showed anti-platelet activity. Four fractions (two from each plant extract) were selected for further sub-fractionation. Fourteen of 35 sub-fractions were selected for further testing of anti-platelet aggregation activity with 12 sub-fractions demonstrating positive antiplatelet activity. Positive sub-fractions were analyzed using liquid chromatography-mass spectrometry to determine their chemical properties. Three compounds that possessed anti-platelet activity were characterized by …

Promising results from the use of problem-based learning (PBL) as a teaching method in medical programs have encouraged many institutions to incorporate PBL into their curricula. This study investigates how applying hybrid-PBL (H-PBL) in a microbiology laboratory impacts students' higher-order thinking as compared to applying a lecture-based pedagogy. The experimental design compared the learning outcomes of two groups of students: the control group and the H-PBL group, for whom PBL cases comprised 30% of the curriculum. Both groups were taught basic skills for the microbiology lab by the same instructor. Using the traditional teaching style for the control group, the instructor offered each student what they needed for their experiments. The H-PBL group practiced experimental design, data analysis, theory proposal, and created research questions by using six study cases that were closely linked to the area of study. The outcome was measured using a pre- and post- assessment consisting of 24 questions that was designed by following Bloom's taxonomy of learning levels. A one-way ANOVA was used to analyze the data. The results showed that for the first three levels of Bloom's taxonomy— knowledge, comprehension, and application—there were no statistically significant differences between the H-PBL and control group gain scores …

Skeletal muscle satellite cells are important for muscle repairing and muscle mass growth. For a successful muscle regenerative process, satellite cells have to sequentially undergoing different stages of myogenic process, i.e. proliferative state and differentiation state. To support this process, the presence of different circulating factors, such as immune cells, cytokines, and hormones, at the appropriate time course is critical. Among these factors, hormones, such as testosterone, cortisol, and IGF-1, have shown to play an important role in satellite cell proliferation and differentiation. Studies investigated the effect of testosterone on satellite cell using a supraphysiological dose in human or in cell culture demonstrated that testosterone is critical in satellite cell myogenic process. Due to the anabolic effect of testosterone on muscle, studies had been focused on the physiological means to increase the circulating testosterone concentration in the body to maximize the muscle mass growth from resistance exercise. The acute and transient increase in testosterone has shown to be beneficial to muscle mass growth and strength gain; however, this change in physiological testosterone concentration on satellite cell myogenesis is not known. Therefore the purpose of this dissertation is to first determine the effect of acute change in exercise-induced hormones on satellite …

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 …

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 …

The accumulation of industrial contaminants in the natural environments have rapidly become a serious threat for human and animal life. Fortunately, there are microorganisms capable of degrading or transforming environmental contaminants. The present dissertation work aimed to understand the genomic basis of microbial degradation and resistance. The focus was the genomic study of the following bacteria: a) Pseudomonas fluorescens NCIMB 11764, a unique bacterium with specific enzymes that allow cyanide adaptation features. Potential cyanide degradation mechanisms found in this strain included nit1C cluster, and CNO complex. Potential cyanide tolerance genes found included cyanide insensitive oxidases, nitric oxide producing gene, and iron metabolism genes. b) Cupriavidus sp. strain SK-3 and strain SK-4. The genome of both bacteria presented the bph operon for polychlorinated biphenyl (PCB) degradation, but we found differences in the sequences of the genes. Those differences might indicate their preferences for different PCB substrates. c) Arsenic resistant bacterial communities observed in the Atacama Desert. Specific bacteria were found to thrive depending on the arsenic concentration. Examples were Bacteroidetes and Spirochaetes phyla whose proportions increased in the river with high arsenic concentrations. Also, DNA repair and replication metabolic functions seem to be necessary for resistance to arsenic contaminated environments. Our …

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 …

Extreme climatic events such as droughts are known to eliminate aquatic biota and alter community structure and function. Perennial headwater springs provide important drought refugia to benthic macroinvertebrates and an important source of colonists via drift or aerial adults to intermittent streams post-drought. During a supra-seasonal drought in North-central Texas summer and fall 2006, benthic macroinvertebrates from persistent groundwater-dependent macrohabitats of varying hydrological connectivity and riparian shading were studied: perennial riffles, connected pools, shaded disconnected pools, and full sun disconnected pools. Riffles were a distinct habitat with significantly higher taxa richness, proportion of lotic taxa, diversity and evenness than other macrohabitats. Macrohabitats were found to be important refugia for 106 benthic macroinvertebrates and 4 microcrustacean taxa. Throughout the extreme drought, perennially flowing habitats were refugia to 19 taxa (17.9% total taxa) not collected in disconnected pools. Shaded disconnected pools contained lotic taxa not previously known to be able to complete their lifecycles in lentic habitats, emphasizing the importance of groundwater effluent and shading. With the resumption of flow at a downstream intermittent site of Ash Creek in mid-October 2006, an annual recolonization study was conducted comparing the perennial headwaters’ benthic macroinvertebrate taxa richness, densities and community ecology with the downstream …

This dissertation has two overall objectives: first, to demonstrate the utility of paleozoological data for ongoing and future mussel-conservation efforts in Texas and second, to evaluate whether simple measures of habitat (e.g., water depth, velocity and particle size) are important for demonstrating the within-habitat spatial separation of mussels. Although these topics may seem disparate, both are important for increasing our understanding of unionid ecology and biogeography. Chapters 1 through 3 examine the use of paleozoological data for mussel conservation. Although these types of data are not new they have rarely been used in mussel conservation efforts within Texas. This is unfortunate because paleozoological data can provide an excellent record of the mussel fauna prior to wide-scale modern impacts and in areas where historical survey data are lacking. Chapter 4 examines whether assessments of microhabitat for mussels using simple measures of habitat (e.g., water velocity, depth and particle size) are useful. Recent studies have suggested that these measures do not explain the mussel distribution in flowing streams. If this is correct, instream flow studies using this approach need to be revised. Results of Chapter 4 indicate that mussels in the lower Brazos River basin are constrained in distribution by the availability …

Thrombocytes are important players in hemostasis. There is still much to be explored regarding the molecular basis of the thrombocyte function. In our previous microarray analysis data, we found IFT122 (an intraflagellar transport protein known to be involved in cilia formation) transcripts in zebrafish thrombocytes. Given recent discoveries of non-ciliary roles for IFTs, we examined the possibility that IFT122 affects thrombocyte function. We studied the role of IFT122 in thrombocyte function. We also found that IFT122 plays a central role in thrombocyte activation initiated by the agonists ADP, collagen, PAR-1 peptide and epinephrine. Although the receptors for ADP, PAR-1 peptide and epinephrine are present in the zebrafish genome, the collagen receptor GPVI was missing. In this study, we identified G6fL as a collagen receptor in zebrafish thrombocytes. Furthermore, IFT knockdown results in reduction in Wnt signaling. The Wnt signaling has been shown to be involved in megakaryocyte proliferation and proplatelets production. Therefore, defects in IFT could lead to thrombocytopenia. Splenectomy is performed in humans to treat such conditions. Therefore, in this study we developed a survival surgery protocol for splenectomy. We have shown that number of thrombocytes and their microparticles increase following splenectomy in zebrafish. Thus overall the studies on …