As the threat of antimicrobial-resistant infections continues to rise, the need for novel antibiotics grows. Targeting virulence factors in bacterial pathogens is one potential strategy for antibiotic development because inhibiting virulence would decrease the ability of the pathogen to evade the host immune response. This strategy may decrease the development of resistance since the treatment is not directly bactericidal and there is less selective pressure put on the bacteria population. Our goal is to discover new virulence genes in Bacillus anthracis that could potentially be a therapeutic target. Specifically, we are interested in finding genes that allow B. anthracis to acquire iron from the host. For bacterial pathogens, iron is critical for growth and often a limiting nutrient in the host. It has been linked with proper functioning of electron transfer proteins and superoxide dismutase enzymes. In B. anthracis infection, iron is acquired from host hemoglobin through a hemolytic pathway, but the complete mechanism of this is unknown. Approximately 1000 transposon mutants of B. anthracis were screened for the inability to acquire iron from hemoglobin, and five were deficient in acquiring iron from hemoglobin in in vitro assays. Of those five mutant strains, only one (9F12) also exhibited an in vivo phenotype using the wax worm model of infection. The gene disrupted in the 9F12 transposon mutant is the dUTPase/aminopeptidase gene. Our aim in this study is to confirm that the disruption of the dUTPase gene leads to the inability to acquire iron from hemoglobin in B. anthracis. Using targeted mutagenesis, we created an insertional mutant strain to disrupt the dUTPase gene and we are currently testing it, along with WT and 9F12, for the ability to grow in iron-limited conditions with or without hemoglobin. Confirmation of this phenotype will demonstrate that the dUTPASE/aminopeptidase gene is important for iron acquisition from hemoglobin and will support further studies to understand the role of this gene in the virulence of B. anthracis.

Aquatic parameters such as increased temperatures and dissolved oxygen levels is critical in determining the survival and ability to thrive of trout species, including the Bonneville cutthroat trout. Bonneville cutthroat trout (Oncorhynchus clarkia Utah), a subspecies of Yellowstone cutthroat trout, originated in the Bonneville Basin and is native to many river basins in Utah, Wyoming, Idaho, and Nevada(Duff 1996). East Canyon Creek is a headwater tributary in the Weber River Basin of northern Utah, and a stream where Bonneville cutthroat trout are native. However, due to the introduction of nonnative trout and multiple causes of habitat quality decline, they no longer occur in the stream. Over the summer of 2018, I participated in data collection which assessed the habitat qualities of East Canyon Creek. This data includes temperature, aquatic, and riparian qualities. Data on the corresponding summer for dissolved oxygen is available as well. When compared to Colorado’s Coldwater Criteria, it appears that the temperatures of East Canyon Creek exceeded the acute (22.1°C) and chronic (17.0°C) upper thermal thresholds for cutthroat trout(Todd et al 2008). When compared to the acute (5.0 mg/L) and chronic (6.0 mg/L) dissolved oxygen minimum concentrations(Null et al 2017), East Canyon Creek’s concentrations appear to have dropped below the identified concentrations. The objective of this paper is to statistically analyze the temperature and dissolved oxygen data on East Canyon Creek from 2018, and determine if a restoration project of Bonneville cutthroat trout in East Canyon Creek would be successful. Through the data analysis, we have found that water temperatures during the summer months have significantly exceeded both acute and chronic upper survival limits, and that dissolved oxygen concentrations are significantly lower than the minimum chronic survival level, indicating that East Canyon Creek is not yet suitable for a successful reintroduction of Bonneville cutthroat trout.

The BRCA1 gene encodes an 1863 amino acid protein that is relevant in many essential biological pathways, most notably DNA damage response and tumor suppression. In many instances, BRCA1's functions depend on interaction with other cellular components. One such binding partner is P53, another important tumor-suppressing protein that cooperates with BRCA1 to inhibit cancer cell growth. However, the nature of this interaction is not yet fully understood. Here we developed a biochemical assay to investigate the exact binding site for P53 in the central domain of BRCA1. The discovery of such binding sites allows future studies to identify the precise amino acid residues involved in binding and better predict the effect of mutations in the binding site on BRCA1's ability to inhibit carcinogenesis.

Breast cancer (BC) is the second most commonly diagnosed cancer among American women after skin cancer. Traditional treatments of BC include surgery, radiation, and chemotherapy therapy; however, these treatments are non-specific and potentially kill peripheral, healthy cells. There emerges a need for more specific treatments, most notably to develop chemotherapy agents that target a unique feature of the cancer cells. Interestingly, 70% of BC cells upregulate estradiol-dependent pathway, a characteristic essential for rapid cell growth. Current BC drugs, such as Herceptin and Tamoxifen, have targeted this pathway to preferentially kill BC cells. However, most women relapse within 15 years due to drug-resistance. Thus, there is a need for new chemotherapeutic drugs. Our research group studies a novel estrogen-receptor targeting drug: Est-3-Melex. This compound has the estradiol molecule linked to a DNA alkylating agent, Melex. We hypothesize that Est-3-Melex enters the cancer cells via an interaction between the estradiol moiety and the estrogen receptor alpha (ER-alpha). ER-alpha then enters the nucleus and binds to Estrogen Response Elements on the DNA. This movement positions the Melex moiety on the DNA and allows the transfer of a methyl group to the N3 adenine on the DNA. In this project, we test the hypothesized mechanism of action of our compound. Since Est-3-Melex has a DNA methylation component (Melex) conjugated to estrogen, our hypothesis is that after the drug binds to the estrogen receptor in the cytosol, it translocates to the nucleus, specifically methylates the N3-region of adenine bases, eventually triggering cell death.

Alzheimer’s Disease (AD) is the 6th leading cause of death in the US. More than 44 million people worldwide, including 5.7 million Americans, are living with this neurodegenerative disease, and those numbers continue to climb. One of the features associated with AD is a disrupted sleep/wake cycle. Sleep is essential for many psychological and biological functions. A reported 35.3% of adults get less than the minimum 7 hours of sleep per night recommended by the National Sleep Foundation. Evidence suggests a bidirectional relationship between sleep loss and AD. Previous research indicates that disruptions in sleep often precede symptoms of AD such as cognitive impairments and memory loss. Chronic sleep loss has been associated with increased amyloid-beta and proinflammatory cytokines in the brain. Extended release of these proinflammatory cytokines can lead to increases in amyloid beta in the brain, which aggregates to form plaques that disrupt neuronal communication, a hallmark of AD. The aim of the present study was to elucidate the interaction between chronic sleep restriction, inflammation, and AD pathology in C57BL6/J mice. Our lab has previously demonstrated that mice administered seven consecutive days of LPS, a bacterial mimetic, exhibit increases in amyloid beta and proinflammatory cytokines in the brain, as well as cognitive deficits. Furthermore, research from our lab has shown that stress can exacerbate the effects of LPS. Healthy C57BL6/J adult mice were subjected to the multiple platform method of sleep disruption for 10 hours per day for 6 weeks. After receiving 7 consecutive days of either LPS or saline injections, animals were subjected to contextual fear conditioning to assess cognitive functioning, and hippocampal amyloid beta levels were quantified. While 7 days of LPS administration did not increase amyloid beta or cognitive deficits in contextual fear conditioning, chronic sleep restriction was associated with deficits in contextual fear acquisition and increased levels of hippocampal amyloid beta compared to control groups. Therefore, chronic sleep loss may have detrimental effects to cognitive function through increasing amyloid beta levels in the hippocampus. Given the large percentage of adults reporting getting less than the minimum recommended 7 hours of sleep per night, combined with the alarming climb in rates of AD and a growing body of work suggesting a link between these trends, investigating the detrimental effects of not getting enough sleep is an essential area of study.