Many high school students demonstrate an overall lack of interest in science. Traditional teaching methodologies seem to be unsuccessful at engaging students \u2014 one explanation is that students often interpret what they learn in school as irrelevant to their personal lives. Active learning and case based learning methodologies seem to be more effective at promoting interest and understanding of scientific principles. The purpose of our research was to implement a lab with updated teaching methodologies that included an active learning and case based curriculum. The lab was implemented in two high school honors biology classes with the specific goals of: significantly increasing students' interest in science and its related fields; increasing students' self-efficacy in their ability to understand and interpret the traditional process of the scientific method; and increasing this traditional process of objectively understanding the scientific method. Our results indicated that interest in science and its related fields (p = .011), students' self-efficacy in understanding the scientific method (p = .000), and students' objective understanding of the scientific method (p = .000) statistically significantly increased after the lab was administered; however, our results may not be as meaningful as the p-values imply due to the scale of our assessment.

This study was conducted as part of an underlying initiative to elucidate the mechanism of action of natural antibacterial clay minerals for application as therapeutic agents for difficult-to-treat infections such as methicillin-resistant Staphylococcus aureus (MRSA)-derived skin lesions and Buruli ulcer. The goal of this investigation was to determine whether exposure to the leachate of an antibacterial clay mineral, designated as CB, produced DNA double-strand breaks (DSBs) in Escherichia coli. A neutral comet assay for bacterial cells was adapted to assess DSB levels upon exposure to soluble antimicrobial compounds. Challenges involved with the adaptation process included comet visualization and data collection. To appropriately account for antimicrobial-mediated strand fragmentation, suitable control reactions comprised of exposures to water, ethanol, kanamycin, and bleomycin were developed and optimized for the assay. Bacterial exposure to CB resulted in significantly longer comet lengths compared to negative control exposures, suggesting that CB killing activity involves the induction of DNA DSBs. The results of this investigation further characterize the antimicrobial mechanisms associated with a particular clay mineral mixture. The adapted comet assay protocol described herein functions as an effective tool to assess double-strand fragmentation resulting from exposure to soluble antimicrobial compounds and to visually compare results from experimental and control reactions.

Staphylococcus aureus and Staphylococcus epidermidis are among the most common causes of hospital-acquired infections5, 7, 8. Despite the advancements in modern antimicrobials, infections from these organisms can be very difficult to treat, and equally as difficult to prevent 6,7. These organisms’ abilities to form biofilms are directly related to their abilities to cause infections. In biofilms, the staphylococcal species can survive antibiotics and immune responses much better than planktonic cells7. Tolaasin—a toxin and natural biosurfactant produced by P. tolaasii—has been briefly tested against biofilm formation, and the results suggested that it could have inhibitory effects. In order to further confirm and expand upon this potentially useful data, additional testing was performed to determine the effects of tolaasin on the two organisms. In addition, laser treatment was tested on E. faecalis in order to supplement our current understanding of biofilm behavior, and provide additional data to suggest alternative agents against biofilm growth.
This thesis addresses the following questions: What are the best methods to test the effects of tolaasin, cephalexin and laser on the biofilms of S. aureus and S. epidermidis? Does tolaasin prevent or disrupt biofilm formation in S. aureus and S. epidermidis? Does tolaasin work synergistically with cephalexin to prevent biofilm growth and maturation in S. aureus and S. epidermidis? And, what effects does laser treatment have on E. faecalis biofilms? In order to answer these questions, tolaasin was isolated from P. tolaasii, and biofilms were pre-treated with tolaasin. Trials were performed with tolaasin, cephalexin, or a combination of both. The effectiveness of each treatment was determined by observing the biofilm growth. The protocols were then optimized and trials were repeated. Additionally, E. faecalis biofilms were exposed to laser treatment. Using confocal microscopy, the biofilms were observed and quantitative results were used to determine the effectiveness of the treatment. Overall, the results indicated that tolaasin has little effect on biofilm growth. However, further investigation is necessary to confirm these results due to some inconsistent data obtained over the course of the trials. Variations and improvements to the protocol are necessary to accurately determine tolaasin’s potential role in healthcare. Finally, the results of the laser trials suggest that EDTA in conjunction with laser treatment could be useful in cleaning root canals and eliminating post-procedural biofilms—thereby preventing infections.

Methane (CH4) is very important in the environment as it is a greenhouse gas and important for the degradation of organic matter. During the last 200 years the atmospheric concentration of CH4 has tripled. Methanogens are methane-producing microbes from the Archaea domain that complete the final step in breaking down organic matter to generate methane through a process called methanogenesis. They contribute to about 74% of the CH4 present on the Earth's atmosphere, producing 1 billion tons of methane annually. The purpose of this work is to generate a preliminary metabolic reconstruction model of two methanogens: Methanoregula boonei 6A8 and Methanosphaerula palustris E1-9c. M. boonei and M. palustris are part of the Methanomicrobiales order and perform hydrogenotrophic methanogenesis, which means that they reduce CO2 to CH4 by using H2 as their major electron donor. Metabolic models are frameworks for understanding a cell as a system and they provide the means to assess the changes in gene regulation in response in various environmental and physiological constraints. The Pathway-Tools software v16 was used to generate these draft models. The models were manually curated using literature searches, the KEGG database and homology methods with the Methanosarcina acetivorans strain, the closest methanogen strain with a nearly complete metabolic reconstruction. These preliminary models attempt to complete the pathways required for amino acid biosynthesis, methanogenesis, and major cofactors related to methanogenesis. The M. boonei reconstruction currently includes 99 pathways and has 82% of its reactions completed, while the M. palustris reconstruction includes 102 pathways and has 89% of its reactions completed.

Studies have demonstrated that viruses such as human immunodeficiency virus [HIV], M13 bacteriophage, and murine cytomegalovirus [MCMV] have been effectively inactivated by exposure to ultra short-pulsed lasers (6,7,10,11,14,15,17). Ultra short pulse laser shows promise as a new method for non-invasive antiviral treatments (17). This method can be used to prevent problems such as drug resistance that is currently rising in numbers. According to the Center for Disease Control [CDC], there are more than two million people in the United States of America that are infected with antimicrobial-resistant infections and at least 23,000 deaths per year occur as a result (19). In this study, ultra-short pulses, specifically Ti-Sapphire Laser [USP Ti-Sapphire Laser] will be evaluated for viral inactivation. The virus chosen for this study was MS2 bacteriophage, which is a non- enveloped, icosahedral, single-stranded RNA [ssRNA] bacteriophages that infects F+ pilus Escherichia coli (16). It was hypothesized that ultrashort pulses from a Ti-Sapphire laser will inactivate MS2 bacteriophage. Inactivation was measured using plaque-forming units [PFU/mL] as an indicator. It was expected that there would be an increase in inactivation of MS2 bacteriophage with an increase in irradiation duration. The results indicated that MS2 bacteriophage was highly sensitive to irradiation treatments of the USP Ti-Sapphire Laser. The concentration of MS2 bacteriophage decreased by 107 PFU/mL after being treated for various time periods ranging from 5 minutes to 150 minutes. Longer duration of USP Ti- Sapphire Laser treatment inactivated more MS2 Bacteriophage.

Collaborative learning has been found to enhance student learning experiences through interaction with peers and instructors in a way that typically does not occur in a traditional lecture course. However, more than half of all collaborative learning structures have failed to last very long after their initial introductions which makes understanding the factors of collaboration that make it successful very important. The purpose of this study was to evaluate collaborative learning in a blended learning course to gauge student perceptions and the factors of collaboration and student demographics that impact that perception. This was done by surveying a sample of students in BIO 282 about their experiences in the BIO 281 course they took previously which was a new introductory Biology course with a blended learning structure. It was found that students agree that collaboration is beneficial as it provides an opportunity to gain additional insight from peers and improve students' understanding of course content. Also, differences in student gender and first generation status have less of an effect on student perceptions of collaboration than differences in academic achievement (grade) bracket.

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related mortality in the USA and throughout the world. Two phenotypes that promote this deadly outcome are the invasive potential of NSCLC and the emergence of therapeutic resistance in this disease. There is an unmet clinical need to understand the mechanisms that govern NSCLC cell invasion and therapeutic resistance, and to target these phenotypes towards abating the dismal five-year survival of NSCLC. The expression of the tumor necrosis factor receptor superfamily, member 12A (TNFRSF12A; Fn14) correlates with poor patient survival and invasiveness in many tumor types including NSCLC. We hypothesize that suppression of Fn14 will inhibit NSCLC cell motility and reduce cell viability. Here we demonstrate that atorvastatin calcium treatment reduces Fn14 expression in NSCLC cell lines. Prior to Fn14 protein suppression, atorvastatin calcium modulated the expression of the Fn14 modulators P-ERK1/2 and P-NF-κβ. Atorvastatin calcium treatment inhibited the migratory capacity in H1975, H2030 and H1993 cells by at least 55%. When chemotactic migration in H2030 cells was induced by the Fn14 ligand TNF-like weak inducer of apoptosis (TWEAK) treatment, atorvastatin calcium successfully negated any stimulatory effects. Inversely, treatment of NSCLC cells with cholesterol resulted in a statistically significant increase in migration. Depletion of Fn14 expression via siRNA suppressed the migratory effect of cholesterol. Finally, atorvastatin calcium treatment sensitized cells to radiation treatment, reducing cell survival. These data suggest that atorvastatin calcium may inhibit NSCLC invasiveness through a mechanism involving Fn14, and may be a novel therapeutic target in NSCLC tumors expressing Fn14.

ABSTRACT
Environmental and genetic factors influence schizophrenia risk. Individuals who have direct family members with schizophrenia have a much higher incidence. Also, acute stress or life crisis may precede the onset of the disease. This study aims to understand the effects of environment on genes related to schizophrenia risk. It investigates the impact of sleep deprivation as an acute environmental stressor on the expression of Htr2a in mice, a gene that codes for the serotonin 2A receptor (5-HT2AR). HTR2A is associated with schizophrenia risk through genetic association studies and expression is decreased in post-mortem studies of patients with the disease. Furthermore, sleep deprivation as a stressor in human trials has been shown to increase the binding capacity of 5-HT2AR. We hypothesize that sleep deprivation will increase the number of cells expressing Htr2a in the mouse anterior prefrontal cortex when compared to controls. Sleep deprived that mice express EGFP under control of the Htr2a promoter displayed anteroposterior gradients of expression across sagittal sections, with concentrations seen most densely within the prefrontal cortex as well as the anterior pretectal nucleus, thalamic nucleus, as well as the cingulate gyrus. Htr2a-EGFP expression was most densely visualized in cortical layer V and VI pyramidal neurons within the lateral prefrontal cortex of coronal sections. Furthermore, the medial prefrontal cortex contained significantly cells expressing Htr2a¬-EGFP than the lateral prefrontal cortex. Ultimately, the hypothesis was not supported and sleep deprivation did not result in more ¬Htr2a-EGFP expressing cells compared to basal levels. However, expressing cells appeared visibly brighter in sleep-deprived animals when compared to controls, indicating that the amount of intracellular Htr2a-GFP expression may be higher. This study provides strong visual representations of expression gradients following sleep deprivation as an acute stressor and paves the way for future studies regarding 5H-T2AR’s role in schizophrenia.

As a major cause of nosocomial infections, biofilms such as those caused by Staphylococcus aureus and Staphylococcus epidermidis pose large concerns in the field of healthcare due to their extreme durability and resistance to treatment. While all biofilms grow similarly in a series of three stages: 1. Adhesion 2. Maturation 3. Dispersal, Staphylococcal species such as S. aureus and S. epidermidis make use of unique growth factors in order to form prolific and durable biofilms. Due to the prevalence and risks associated with bacteria, many antibacterial methods have been created to treat bacterial infections. Although many antibacterial methods exist, there is still a great need for additional and more effective methods to treat and prevent serious bacterial infections associated with biofilm growth, because incidences of bacterial infection and resistance, especially in medical settings, are on the rise. In recent research, the exotoxin tolaasin, produced by the bacterium Pseudomonas tolaasii has briefly been shown to exhibit antibacterial effects. Based on previous research and tolaasin's observed pore forming and detergent properties, it is hypothesized that tolaasin will disrupt and prevent staphylococcal biofilm growth either independently or synergistically with existing antibiotics. If this is confirmed, tolaasin may have major implications within the future of healthcare, particularly in the field of antibiotics. In order to optimally use tolaasin as an anti-biofilm agent, potential anti-biofilm applications would aim to prevent and treat biofilm infections at the most common sites of biofilm growth such as catheters, medical instruments, implanted medical devices, and surgical sites. In addition, under the assumption that tolaasin will be found effective in inhibiting biofilm growth and infection, this thesis proposes future anti-biofilm technologies that could use tolaasin as an anti-biofilm agent in order to prevent biofilms and associated infections. While there are many potential and promising ways that tolaasin could be used as an anti-biofilm agent in the future, there are still possible limitations that would need to be investigated through further research before these applications can come to fruition. Ultimately, if future research successfully determines that tolaasin can be used to make anti-biofilm technologies that are biocompatible, durable, and effective, then technologies using tolaasin as an anti-biofilm agent may more effectively ensure sterility of medical devices and prevent bacterial biofilms and infections, and may eventually save lives.

Abstract Development of a Vaccine for Immunization Against Smallpox and Anthrax Jason Maurice Cameron Biological weapons are often considered to be the most dangerous weapons of mass destruction because of there potential to infect huge numbers of people, who may then in turn infect others who were not even present at the point of initial impact. Among the most feared biological weapons are those that contain smallpox and anthrax because of these diseases' high rates of both infection and death. For this reason, the development of a vaccine that immunizes the receivers against both smallpox and anthrax would be great progress. This study seeks to develop such a vaccine by constructing a recombination plasmid that will introduce new genes that combat anthrax into the strain of vaccinia virus (VV), the virus used to vaccinate against smallpox. This study includes a highly detailed analysis of the various processes used to attempt this recombination and proposes plans further research into the subject.