Bacillus anthracis is a gram-positive, spore-forming bacterium and the causative agent of the deadly disease anthrax. The B. anthracis genome consists of chromosomal genes and the pXO1 and pXO2 plasmids that strongly contribute to the bacteria’s deadly nature. While the virulence factors associated with the plasmids have been extensively studied, we believe there are still undiscovered chromosomal genes that may also have important virulence factors. To identify novel chromosomal genes associated with B. anthracis virulence, we screened a transposon mutant library of B. anthracis Sterne strain for increased sensitivity to reactive oxygen species. Reactive oxygen species, such as hydrogen peroxide, have many functions in mammalian immune defenses and wild type B. anthracis is able to subvert this host defense. Sensitivity to reactive oxygen species was tested through in vitro hydrogen peroxide assays and after several rounds of screening, eight mutants were confirmed as susceptible. We next tested whether any of these mutants were attenuated in vivo using our invertebrate animal model, Galleria mellonella and found several mutants with decreased virulence. We are currently working on determining the location of the transposon insertion to find which chromosomal gene is disrupted. This could lead to the discovery of novel B. anthracis virulence genes and eventually possible treatment targets for future anthrax outbreaks and attacks.

The swim performance assay is a behavioral assessment used to measure cardiovascular function in fish. Previously, the laminar flow assay (LFA) has been the standard method of assessing swim performance in adult fish to measure their cardiac output. The spinning task assay (STA) is a novel, accessible method of assessing swim performance; however, previous studies have not compared the two methods. Additionally, there is little documentation of swim performance in larval fish, a more sensitive study subject for toxicological research. Therefore, the aim of this research is to compare the swim performance of fish in the LFA to those in the STA to determine which method is better for assessing swim performance in larval fathead minnows (Pimephales promelas). In this study, the percent of fish that fail to swim in the LFA is inversely proportional to the age of the fish, but in the STA, there is no correlation between percent failure and fish age. Results show that as fish increase in size, swim performance in the LFA improves, making it a more representative, predictable assay. Results from the STA indicate that swim performance in fish does not improve with size and performance in the STA is not correlated with performance in the LFA. Ucrit values from the LFA have less variation than those from the STA. The results of this study show that the LFA is a more suitable modality for assessing swim performance in larval fathead minnows.

Porous silicon nanoparticles exhibit great potential as drug delivery vectors due to their high surface-area-to-volume ratio allowing for increased efficacy of surface functionalization and therapeutic loading capabilities. This data set demonstrates the fabrication of a class of plant-derived materials which are sub-micron in size and capable of functionalization with primary amine groups through the addition of APTES.
The production of porous silicon particles (pSi) is achieved through magnesiothermic reduction of silica containing Tabasheer powder isolated from the nodal joints of the Bambuseae plant. Efficacy of this reduction is evaluated using techniques including X-ray diffraction and Energy-dispersive X-ray spectroscopy which show successful reduction of silica starting material to porous silicon.
High energy ball milling followed by reduction is used to produce pSi particles of sub-micrometer size while also allowing for a significantly higher yield (~90%) of material than previous methods. Particle size is confirmed via electron microscopy and dynamic light scattering (DLS).
Following reduction, surface functionalization of silicon nanoparticles with primary amine groups was carried out using a 4% (v/v) solution of APTES in acetone. The evaluation of this functionalization was conducted using techniques including zeta potential and infrared spectroscopy (IR). Zeta potential values are found to be approximately -10 mV. This data demonstrates successful amino silanization.
The results achieved through these methods suggest successful fabrication of pSi nanoparticles and subsequent functionalization for future use as a drug delivery vector.

Drug delivery is the process by which medications are administered to the body. This is complex due to the difficulty of determining compounds that have the proper biocompatibility and permissibility to our human cells. Many medications are taken orally; however, there are advantages to administering medication subcutaneously or by inserting it in the inner corner of the eye. Porous films made out of biocompatible polymers provide a good platform for drug delivery as they have the ability to be loaded with plant derived porous Silicon. Functionalizing the porous silicon using (3-aminopropyl)triethoxysilane and glutaraldehyde can be done in an attempt to covalently attach particles to the film which is important for embedding them into the pores of the film. Porous silicon has biocompatible properties and can be loaded with drugs then modified to alter the release of those drugs in the body. This method has the potential to be a useful drug delivery method due to the biocompatible and biodegradable properties of the material and the ability to manipulate the material in order to maximize drug release.

The objective of this project is to make a vaccine that will negate the effects of the powerful opioid fentanyl in the long term. Fentanyl is a strong synthetic opioid that is 50 to 100 times more potent than morphine. According to the CDC, there were over 70,000 deaths due to street drug overdoses, which has increased in the last ten years. 40 % of these deaths are related to fentanyl overdoses, therefore it is imperative that approaches are developed to combat this alarming increase in deaths. The vaccine against fentanyl will be synthesized out of molecules that will take advantage of fentanyl’s amide functional group to be hydrolyzed into safe byproducts. Any patient that is administered with the vaccine, will not feel the effects of the opioid because the immune system will hydrolyze the drug as soon as it enters. This project will exploit the properties of both catalytic antibodies (CAbs) and transition state analogs. The Cabs will trigger an immune response to attract phagocytic cells, such as macrophages to phagocytose pathogens and eliminate them from the system. However, if the molecule resembles the transition-state of fentanyl hydrolysis, then the antibodies can cleave the fentanyl in a fast and efficient manner due to their catalytic properties. Therefore, after immunization, a person who is addicted to fentanyl would no longer feel the effects of the opioid because it will be degraded as an immune response is triggered, creating a long-term possible solution to one factor of the “opioid crisis.”