Open Research Oklahoma (Oklahoma State Univ.)
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Magnetically assisted colorimetric sensing of insulin and oligonucleotides in biofluids
No full textColorimetric approaches capable of detecting ultra-low concentrations of specific molecular targets in real sample matrices hold immense significance as point-of-need biosensors, especially in the post-pandemic era. To address this critical need, my dissertation research focuses on a magnetic particle-based colorimetric immunosensor for the picomolar detection of serum insulin by a two-antibody (capture and detection antibodies) sandwich assay format, which offers high selectivity to insulin. Colorimetric detection occurs through the enzymatic oxidation of a chromogenic substrate by the horseradish peroxidase-labeled detection antibody within the immuno-assembly. This method is applicable for detecting insulin in both type-I diabetes and healthy serum samples. Extending the protein immunosensor platform, we have developed a hybridization colorimetric sensor capable of detecting recombinantly synthesized RNA sequences of the SARS-CoV-2 virus as target molecules spiked in undiluted serum and saliva. To enhance sensitivity and further lower detection limits, we have integrated the colorimetric hybridization sensor with an electrochemical transducer for RNA detection and a surface plasmon transducer for small molecule detection
Creating microvasculature using 3D printing of hydrogels
No full text3D printing has gained significant interest in recent years as a tool in additive manufacturing. Within the medical field, bioprinting has been used to develop synthetic tissues. Despite significant advances in 3D bioprinting technologies, there is a lack of understanding of various factors such as nozzle diameter in relation to cell size, nozzle length and flow rates. One other barrier is the inability to create microvasculature that exists within our tissues, to be able to go below 0.1 mm diameter during bioprinting. Printing cell-laden structures is also not widely tested, as in most cases, a fiber is printed and then cells are randomly introduced. In this study, I proposed a chitosan-gelatin hydrogel and first printed a single fiber to test how it affects cells. I investigated different concentrations of the hydrogel mixture. Different nozzle sizes were tested with the cells, analyzed them for cell area, and shape. Also, I used previously obtained data to show how nozzle size to cell size ratio affects cell morphology. Next, I created hollow fibers with 0.1 mm that can be used to form microvasculature network by using a custom-made coaxial nozzle. Using the factorial design of experiments, I tested the effect differing printing speeds and nozzle lengths, reaching a repeatable and optimal process for creating fibers with average dimensions of < 0.1 mm. I used liver sinusoidal endothelial cells during printing to mimic blood vessels. Cells survived the printing process even after 5-7 days incubation, both within the core and the shell. Although fibers were stable, there was very little cell spreading. I evaluated a different alginate-based hydrogel to check the usability of the coaxial nozzle. Every other parameter was kept the same. The alginate-gelatin mixture showed a significant increase both in cell spreading, but also in size
Improving epidemiological surveillance algorithms for Francisella tularensis using mass spectrometry and proteomics
No full textThis dissertation investigates the genomic and proteomic characterization of Francisella tularensis isolates using advanced bioinformatics tools and mass spectrometry techniques to enhance pathogen identification, epidemiological surveillance, and public health response. Whole genome sequencing (WGS) was employed alongside tools such as CanSNPer2, CARD, and RAxML to classify F. tularensis canSNPs, detect antimicrobial resistance, and understand the genetic diversity of isolates primarily collected from Oklahoma and North Carolina. The study achieved a 95% classification accuracy in determining canSNPs, with antibiotic resistance genes vanYB and FTU-1 identified across all isolates. These findings underscore the utility of WGS in microbial forensics and outbreak management.
Additionally, this research explored the application of Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) for comprehensive proteomic analysis of F. tularensis isolates. LC-MS/MS identified differential protein expression between subspecies, with proteins such as isochorismatase-like domain-containing protein and indolepyruvate decarboxylase showing higher expression in F. tularensis subsp. tularensis. The study also demonstrated that machine learning models, specifically support vector machines (SVM), could classify F. tularensis isolates with 98% accuracy using proteomic data, highlighting the potential of LC-MS/MS in pathogen classification.
Furthermore, phenotypic analysis using Biolog Omnilog revealed metabolic diversity across isolates, emphasizing the value of combining genomic and phenotypic data for a comprehensive understanding of bacterial behavior. This research contributes to public health and biodefense strategies by providing rapid identification tools for bacterial biothreat agents, improving pathogen detection and classification in clinical and field settings. The findings have significant implications for future research in bacterial genomics, proteomics, and epidemiology, suggesting that LC-MS/MS is a promising tool for enhancing the accuracy of microbial diagnostics
Investigating administrator and instructor perceptions of employability skill development in Oklahoma technology centers
No full textThis study used a mixed methods approach to investigate the perceptions of administrators and instructors of full-time CTE programs in Oklahoma technology centers regarding their ability to develop employability skills in students. This study was framed by the human capital theory which is described as the acquisition of a person’s knowledge and skills (Schultz, 1971). Employability skills have been shown to be one area where human capital has the potential to be developed in students through education and training (Robinson & Garton, 2008). The Employability Skills Framework of Career and Technical Education (CTE) outlines employability skills that should be developed through CTE program participation for college and career readiness (U.S. DOE OCTAE, n.d.). Since the last installment of the Carl D. Perkins Act, there has been a call for CTE reform (Advance CTE, 2021; Jocson, 2018). This reform consisted of four points of focus in the Perkins Act blueprint: “alignment between CTE programs and the labor market; collaboration among secondary and postsecondary institutions, employers, and industry partners; accountability for academic outcomes and employability skills for all students; and innovation to support CTE implementation with effective practices” (Jocson, 2018, p. 641). The purpose of the study was to investigate administrator and instructor perceptions of Oklahoma Career and Technology Education’s capacity to develop employability skills in students to meet the needs of the local community. The perceived ability of full-time CTE programs in technology centers to develop employability skills in students was investigated using a convergent parallel mixed methods design which included a quantitative questionnaire and one on one interviews. A two-article format was used to present findings, conclusions, and recommendations for both administrators and instructors. Overall, administrators perceived a high rating of their programs’ ability to develop these skills than instructors. Problem solving and teamwork have the most ability to be developed while money management is seen to have the least ability to be developed. There was an overarching theme that administrators have a philosophy of technical skills first, while instructors value holistic student development (Dewey, 1938; Prosser & Allen, 1925)
Help kids make friends: Use emotion coaching
No full textThe Oklahoma Cooperative Extension Service periodically issues revisions to its publications. The most current edition is made available. For access to an earlier edition, if available for this title, please contact the Oklahoma State University Library Archives by email at [email protected] or by phone at 405-744-6311
Passive treatment of mine drainage and the behavior of manganese
Get PDFNon-point source pollution poses a significant threat to the environment and can be difficult to prevent and remediate. One contributor to this is mine drainage, which can contain numerous contaminants of concern. Passive treatment systems have been developed as a method of remediating mine drainage. The behavior of manganese within these systems has raised questions regarding its ability to be retained by the organic substrate used for remediation, with a prior study showing manganese sometimes being released in higher concentrations than when it entered the system. The goal of this current study is to further research on the behavior of manganese and determine if there is a pattern of its release within treatment systems. For this study, cubitainers are set up containing an organic substrate and a manganese solution. Biweekly testing is currently being conducted to measure pH, dissolved oxygen, conductivity, and oxidation-reduction potential. Along with this, samples of water from each container are being analyzed for manganese concentrations. This is an ongoing project, and samples will continue to be taken into the next academic year. At the end of the semester, three of the containers will be sacrificed to analyze the manganese content within the organic substrate itself. This information will be used to determine in which part of the substrate the manganese is retained. This will provide further insight into improving passive treatment systems for a variety of contaminants.Environmental Scienc
Effects of β-funaltrexamine on lipopolysaccharide-induced inflammation and behavior
No full textInflammation plays a crucial role in the development, progression, and severity of many prevalent systemic and neurological pathologies. A growing body of evidence has found that inflammation is a key aspect of cardiovascular disease, autoimmune disorders, cancer, neurodegenerative diseases, and neuropsychiatric conditions. Consequently, there is a critical need to develop targeted pharmaceuticals to mitigate systemic inflammation and inflammation-induced behavioral deficits. Our lab has previously demonstrated that acute β-FNA has selective anti-inflammatory effects in normal human astrocytes and reduces systemic inflammatory mediators and anxiety- and sickness-like behavior in male mice, predominantly through inhibition of p38 MAPK and NF-κB. The purpose of this study was to extend our knowledge of the cell-specific effects of acute β-FNA treatment using microglial cells, as well as study chronic pre-treatment effects on systemic inflammation and inflammation-induced behavioral deficits using a preclinical model. BV2 murine microglial cells were co-incubated with β-FNA and LPS at various times, while osmotic drug pumps were surgically inserted into mice that continually dispensed β-FNA for six days prior to LPS injection. Depressive-, anxiety-, and sickness-like behaviors were measured 24h after LPS injection, and then mice were terminated, and plasma, frontal cortex, hippocampus, and spleen tissues were harvested. CCL2, CXCL10, and IL-1β levels were measured in cell supernatant, CCL2 and CXCL10 were measured in frontal cortex, hippocampus, and spleen tissues, and IL-1β and corticosterone were measured in mouse plasma via ELISA. IDO1 and NLRP3 transcription were quantified via RT-qPCR from whole cell lysates and the afore-mentioned tissue homogenates. Additionally, p38 MAPK and STAT1 were quantified via Western blots in whole cell lysates. Acute β-FNA selectively reduced CCL2 and paradoxically potentiated LPS-driven IL-1β in BV2 cells, while exhibiting no effect on p38 MAPK, STAT1, NLRP3, or IDO1. Behavioral deficits were predominantly correlated with neuroinflammatory levels of CCL2, CXCL10, NLRP3, and IDO1. Additionally, chronic β-FNA pre-treatment greatly reduced systemic levels of CCL2 and CXCL10 and inhibited or abolished anxiety- and sickness-like behaviors. Interestingly, β-FNA increased frontal cortex inflammation and anxiety-like behavior under control conditions. Consequently, these studies provide additional insight into the actions and therapeutic potential of β-FNA and underscore areas that need further investigation
Effects of paternal deprivation on behavior, sexual traits, and the stress response in a biparental bird
No full textIn species with biparental care the deprivation of one parent can cause decreases in the quantity of parental care, which can have implications for offspring growth and development. Much of what we know about parental deprivation comes from maternal deprivation studies. However, little is known about the implications of paternal deprivation on the development of offspring behavior, the stress response, and secondary sexual traits. I tested the effects of paternal deprivation in captive zebra finches (Taeniopygia guttata) by removing the male parent at either hatching or fledging and measured the subsequent effects in offspring. In Chapter II of my dissertation, I measured offspring nestling and adult baseline corticosterone and adult isolation-induced corticosterone. I also quantified the effects of paternal deprivation on parental care and offspring growth. I found that single female parents increased some, but not all, behaviors to match the cumulative maternal and paternal behaviors in biparental nests, nestling growth was not affected by paternal deprivation, and offspring that experienced paternal deprivation from hatching had higher corticosterone levels relative to their baseline corticosterone levels after experiencing social isolation. In Chapter III, I quantified the social behavior of adult offspring. I found that offspring raised without paternal care from hatching exhibited a higher number of aggressive behaviors in response to novel, same-sex conspecifics compared to offspring raised by both parents. For Chapter IV, I focused on two secondary sexual traits important to male zebra finch fitness, beak color and song. I found that male offspring raised without paternal care from fledging had less red saturated beaks compared to biparentally raised male offspring. Male song complexity was not affected, but biparentally raised male offspring were less likely to produce birdsong than sons experiencing paternal deprivation. The results from this research indicate that loss of paternal care can impact a variety of traits important to offspring fitness
Mesoscale modeling of microstructural evolution of friction stir welded austenitic stainless steel
No full textFriction stir welding (FSW) is a solid phase processing (SPP) technique allowing for the fusion of metals through means of frictional heat. During friction stir welding, a rotating tool plunges into the material causing localized heating while moving along a path in the metal. A joint weld is created with new microstructural and mechanical properties, differing from the base material. FSW is a favorable technique over conventional welding due to the improved properties produced because the material is not melted, eliminating solidification defects. An increased interest in the process of friction stir welding of austenitic stainless steels has emerged, requiring more modeling and experimentation. Within this thesis, phase field modeling of friction stir welded 304 stainless steel and its resulting microstructure will be analyzed. Typically, friction stir welding is modeled by means of dynamic recrystallization due to the strain the metal is put under during the process. However, within this analysis, friction stir welding will be modeled by means of static recrystallization by simulating grain growth with discrete nucleation where the initial grains will have an assumed amount of deformation energy. The model considers the interfacial energies of grain boundaries, bulk free energy of the free energy change, and stored energy contributions ensuring that recrystallized grains are lower in energy than deformed grains. The model results demonstrate the effect of dislocation density (deformation energy) and temperature have on nucleation and grain growth of recrystallized grains
Modeling recurrent turbidity impairments in Oklahoma and the impact of targeted best management practices on sediment concentrations
No full textThe Clean Water Act mandates that states must maintain a water quality standard to ensure surface waters remain functional for aquatic life and safe for agricultural or recreation use. Water Bodies that exceed any of the defined standards must be reported as threatened or impaired semiannually to the USEPA per section 303(d). If listed, states must then develop and implement a watershed plan for the threatened or impaired waterbodies to reduce pollution and hopefully delist the waterbody. While the waterbody is impaired, states have access to funds set aside by Section 319 that provide funding for monitoring efforts and technical assistance and cost sharing for landowners. Management of non-point source pollution, such as turbidity, is achieved by implementing best management practices (BMPs), which are costly to install and maintain, making it imperative that they provide a lasting reduction in pollution. Turbidity is a leading impairment that largely stems from non-point source pollution, which are sources without easily identifiable outlets. This research analyzed Oklahoma’s 303(d) lists from 2002-2022 for recurrent turbidity impairments. Three case study watersheds were selected to determine potential causes for the list-delist-relist pattern. The Wewoka, Turkey, and Stillwater Creek watersheds were selected based on size, ecoregion, and documented management efforts. The watersheds were then modeled for the 21-year period using the Hydrologic and Water Quality System (OK.HAWQS). A baseline model was run to analyze historic sediment contributions in each watershed, which allowed us to investigate the impact of different land use types on sediment loss and find variations across watersheds in erodibility. The Grassland, Winter Wheat, and Pasture land uses in the Stillwater, Turkey, and Wewoka watersheds lost the most sediment, respectively. BMP scenarios were run for the land use contributing the most sediment in each watershed. Based on the most commonly implemented practices in Oklahoma, we chose enclosure fencing, conservation grazing, and no-till for Stillwater, Turkey, and Wewoka, respectively. These scenarios showed conservation grazing has a positive impact on sediment reduction, up to 50%, while no-till and fencing had a small to neutral impact on sediment loading. These results can be used to inform future management project ad assess the adequacy of current management techniques in a changing climate