This research investigates the effectiveness of HOPE Connection 2.0, a revised version of the therapeutic summer camp developed by The Karyn Purvis Institute of Child Development (KPICD) at Texas Christian University (TCU). Focused on Trust-Based Relational Intervention (TBRI), the camp aims to address the needs of vulnerable children and their families. Through a self-report survey administered to participating caregivers, the study evaluates lasting behavioral and relational developments in the family following their camp experience. The research question explores whether HOPE Connection 2.0 optimally benefits families and how it can be enhanced. Objectives include identifying the most beneficial aspects of the camp, suggesting design improvements, and assessing the reception of TBRI teachings by caregivers and children. The findings aim to inform future iterations of the camp, facilitating continuous improvement and adaptation to better serve participants.

Previous research finds that testosterone serves a major role in psychological and physiological preparedness in competitive environments, with higher testosterone predicting better competitive endurance and overall performance. Despite the performance benefits afforded by testosterone production, high testosterone has also been found to be physiologically costly due to suppressing immune function in the short-term and having detrimental health impacts in the long-term. However, no work has experimentally examined whether men and women’s testosterone response is sensitive to the relative costs and benefits of investing in testosterone production. To address this question, participants were asked to compete in a novel task and were randomly assigned to compete for a $5 (low-value reward) or $20 gift card (high-value reward). Additionally, participants were informed that the winner of the reward would be selected based on their performance (effort-based) or would be randomly selected (random-based). Saliva samples were collected before and after the competitive task and assessed to measure changes in free testosterone levels pre- and post-competition. Results revealed that participants in the high-value reward condition produced more testosterone than those in the low-value reward condition. Similarly, participants in the effort-based winner condition had higher testosterone production than those in the random-based winner condition. These findings suggest that physiological endocrine responses are sensitive to psychological experiences which impact the costs and benefits of testosterone production.

Frustration is a negative emotion that occurs when an organism encounters a reward that is of lesser quality or quantity than expected. To research this emotion in animals, we use a paradigm known as consummatory successive negative contrast (cSNC). In this task, rats are exposed to a high concentration sucrose reward and then downshifted to a lower concentration. In response to this downshift, rats inhibit consumption and reject the downshifted reward beyond that of unshifted controls. This bout of negative emotion is also accompanied by the natural release of endogenous opioids, which modulate dopaminergic activity in the brain. Previous research has shown that when opioid receptors are blocked, frustration is enhanced after a 32-4% sucrose downshift. This experiment aimed to further explore this effect using the drug naloxone, which blocks opioid receptors, and different degrees of sucrose downshifts to determine if naloxone will show the same effect with a less extreme 16-4% sucrose downshift. To test this, rats were trained with 32, 16, or 4% sucrose for ten sessions with all animals then receiving 4% sucrose for the next four sessions along with injections of either 2 ml/kg naloxone or saline. Preliminary data suggest a slight decrease in consummatory behavior after naloxone exposure in both downshift-exposed groups compared to saline controls. This suggests that opioid blockers may play a role in augmenting frustration at varying degrees of downshift, inhibiting the ability of the dopaminergic system in the brain associated with reward and sucrose intake.

A relatively novel area of research in social psychology, existential isolation, can be described as feeling that you are alone in your experience of the world, and that others do not share your perspective or can even come close to understanding it. Existing research finds that existential isolation is related to greater death-thought accessibility, depression, and anxiety and is higher among individuals in non-normative groups (e.g. racial/sexual minorities, low socioeconomic status, etc.). However, the totality of how existential isolation effects mental wellbeing, particularly with regards to our self-perception, remains somewhat unclear. The present research studies aimed to examine the association between feeling existentially isolated and different mental wellbeing outcomes, hypothesizing that higher levels of existential isolation would be associated with poorer self-perception. Using self-report survey measures with samples of 302 undergraduate students (Study 1) and 200 MTurk adults (Study 2,) results revealed that higher levels of existential isolation were positively correlated with greater levels of self-dehumanization, rejection sensitivity, and loneliness, but negatively correlated with greater levels of self-esteem and coping self-efficacy. That is, the results suggest that individuals who feel chronically existentially isolated from others in their daily lives may also develop a more negative self-image and view themselves as less able to handle the stressors they may encounter. Overall, these findings provide preliminary evidence of an association between feeling existentially isolated and poor mental wellbeing outcomes with regards to how an individual views themselves and their capabilities. The implications of these findings for the long-term wellbeing, both physical and mental, of existentially isolated individuals, as well as for their social relationships, will be discussed.

Title: Adverse Childhood Experiences (ACEs) & Academic Performance
Authors: Janae Russell, Casey Call, Ph.D.

College students face many challenges when it comes to discovering their identity, establishing independence, developing a routine, adjusting to a new lifestyle, handling stress and social interactions. Furthermore, students that have been exposed to chronic Adverse Childhood Experiences (ACEs) can experience negative impacts on their mental health and academic performance. ACEs are childhood experiences of emotional, physical, and sexual abuse, and/or household dysfunctions such as substance abuse, mental illness, suicide, or incarceration (Felitti et al., 2019). Felitti et al. (1998) found that ACEs are linked to many different health risk factors such as declining health, smoking, alcohol abuse, depression, and substance abuse. College students that have been exposed to ACEs are more likely to struggle with their mental health, depression, and anxiety (Hatton-Bowers et al., 2023). The purpose of this research is to examine how ACEs impact a college students’ performance. I would like to learn how students define academic success and if they believe they fit that definition. I would like to discover how ACEs influence students’ overall ability to learn and retain information, focus on academic work, develop study habits, and have successful social interactions with peers. I theorize that students who have been exposed to four or more ACEs will struggle more with the demands and pace of college life than students who have 3 or fewer ACEs. This research is essential for universities to be able identify students with ACEs to better understand and support these students.

Felitti, V. J., Anda, R. F., Nordenberg, D., Williamson, D. F., Spitz, A. M., Edwards, V., Koss, M. P., & Marks, J. S. (2019). REPRINT OF: Relationship of Childhood Abuse and Household Dysfunction to Many of the Leading Causes of Death in Adults: The Adverse Childhood Experiences (ACE) Study. American Journal of Preventive Medicine, 56(6), 774–786. https://doi.org/10.1016/j.amepre.2019.04.001

Hatton-Bowers, H., Lombardi, C. M., Kemp, B., Decker, K. B., Virmani, E. A., Brophy-Herb, H. E., & Vallotton, C. D. (2023). Risks and resources for college students’ mental health: ACEs, attachment, and mindfulness. Journal of American College Health, 71(5), 1510–1521. https://doi.org/10.1080/07448481.2021.1942007

Over the past decade, mindfulness-based practices have gained popularity in the mainstream media as a way of alleviating stress. Indeed, state- and trait-based mindfulness is correlated with enhanced well-being, meaning in life, and life satisfaction. In this way, mindfulness-based interventions could be a pivotal tool for educators and colleges aiming to improve student outcomes. The purpose of the current study was to test the feasibility and engagement of a brief mindfulness intervention for nursing students. Participants (N = 72) were recruited from the nursing department at Texas Christian University. Participants were randomly assigned to complete a mindfulness exercise or a relaxation exercise, and then asked to complete survey questions. Engagement in the intervention will be assessed using recruitment numbers and program completion. Additionally, we are expecting that people assigned to the mindfulness exercise will report higher levels of happiness, well-being, and gratitude when compared to those assigned to the relaxation exercise. The implications of this work will be discussed.

The preterite and imperfect past tenses, which do not have exact English equivalents, exemplify grammatical nuance in the Spanish classroom. These nuances evoke questions regarding effective ways to present this material to students. Some evidence suggests that interleaved schedules of practice – in which material is presented in a mixed order during learning – benefit learning of grammar in a non-native language (Nakata & Suzuki, 2019; Pan et al., 2019); however, other evidence suggests that blocked schedules of practice benefit pronunciation learning (Carpenter & Mueller, 2013). Given these mixed outcomes, I compared blocked and interleaved schedules of practice on learning of the preterite and imperfect tenses in Spanish. Participants were randomly assigned to interleaved or blocked practice, in which they classified verb constructions as imperfect or preterite (when conjugated to Spanish) and were given feedback following each item. Participants then completed multiple tests assessing their grammar learning. The interleaved and blocked groups did not significantly differ in their test performance; however, both groups showed significantly improved performance compared to a pre-test, indicating that learning did occur. These outcomes can inform pedagogical practice. Future research should consider time processing feedback, and extend these outcomes to Spanish language learners, with materials translated into Spanish.

With the rate of children growing up in single-parent households on the rise, the challenges that these children experience are becoming more evident. One challenge that often remains unacknowledged is parentification. Many single parents engage in this process due to the number of stressors that they must face on their own. Parentification allows parents to place responsibilities on their children that they would otherwise not do in order to relieve some sort of stress. These responsibilities can include caring for siblings, being a translator, taking on a parent's emotional turmoil, and even paying bills. With this adornment of responsibility, these children can garner mixed emotions about themselves and their purpose in the world. However, we hypothesize that children who experience parentification will have high self-efficacy. The findings of this study may lead to implications that assist single parents in performing positive and sensitive parenting behaviors in order to promote a secure attachment with their children, as well as promote conversations on policy interventions that relieve unique stressors for single mothers so that they avoid having to rely on parentification.

The philosophical concept of free will is often highly debated. Benjamin Libet, an academic in experimental philosophy, discovered through recordings of cerebral activity that there is neural activity that correlates with a decision prior to our conscious declaration of that decision (Libet et al., 1983). In songbirds, previous studies have begun to show an increase in neuronal and respiratory activity in the seconds prior to song production, indicating preparatory action before performing this learned behavior (Daliparthi et al., 2019). In this study, the preparation for the learned motor behavior (song production) is compared against an unlearned control (defecation) in Zebra Finches. Electromyography (EMG) of respiratory muscles is employed as an additional technique to provide more detailed exploration of preparatory motor activity compared to previous studies. Our analysis of EMG recordings focused on the six expirations that occur prior to song production, because previous research has shown that there is an acceleration of the respiratory rhythm occurring in the last three respiratory cycles before song (Méndez et al., 2022). By measuring the electrical activity in the muscle, we hope to provide a more detailed understanding of how birds prepare for their upcoming song. Overall, this study aims to explore motor responses to determine whether expiratory muscular activation is preparatory for and predictive of an upcoming behavioral event. The larger goal of this study is to be able to “read a bird’s mind” by establishing physiological models for predicting behavior before the decision has occurred.

Shifts in mood across the menstrual cycle have been widely explored. However, few researchers have sampled participant experiences more than once or twice in each phase of the cycle. This methodology has limited our understanding of a very heterogenous cycle. We build on previous work by a) increasing sampling frequency and by b) examining how different subphases of the cycle correlate with participant mood. Female participants in the current study were asked to fill out a questionnaire about their mood and experiences every other day throughout their cycle. Participant responses were averaged within the luteal and follicular phases respectively in order to examine significant changes in mood from one half of the cycle to the next. Additionally, participant responses were averaged within each subphase of the cycle (i.e., early follicular, ovulatory, early luteal, and late luteal) in order to examine significant changes in mood within each phase. Results will offer an in-depth analysis into the different shifts in mood that natural-cyclers may experience across the menstrual cycle.

Unhealthy eating behavior plays a major, preventable role in many chronic health conditions, such as obesity, which is a leading cause of early morbidity in the United States. Recent research has focused on the influence of social connections on food choice through the enforcement of food-related social norms that motivate healthy eating practices. While this research illuminates the relationship between social connection and the promotion of healthy eating habits, less is known about how lack of social connectedness (i.e. isolation) may influence eating behavior and food choice differently. Just as there are many ways to be socially connected, there are also many ways to be disconnected. One such form of disconnection is the experience of existential isolation, which is characterized by the feeling that one is alone in their experience of the world. Existing research has found that chronic existential isolation is associated with poor mental wellbeing, such as heightened feelings of loneliness, death through accessibility, depression, anxiety, and reduced self-esteem. Yet, the associations existential isolation may have with physical health outcomes or behaviors remain unclear. The present research aims to investigate the links between feelings of existential isolation and self-reported eating behavior and healthy eating intentions. Across two survey studies, undergraduate students reported their trait levels of existential isolation, loneliness, and healthy eating behavior, with Study 1 assessing food quality, calorie content, and portion size planning, while Study 2 focused on intentions to eat healthy in relation to existential isolation. The results revealed significant negative correlations between existential isolation and self-reported healthy eating behavior, food quality planning, caloric intake planning, portion size planning (Study 1), and healthy eating intentions (Study 2). These associations also remained significant even when controlling for individual differences in interpersonal loneliness. The findings offer preliminary evidence of an association between daily feelings of existential isolation and unhealthy eating cognitions, with more existentially isolated individuals reporting less mindful eating behaviors and intentions. These results highlight the potential role that feeling existentially isolated may have on dietary choices, and subsequent long-term health outcomes. Implications of these findings for future research examining the relationship between social connectedness, existential isolation, and long-term health outcomes related to eating cognition will be discussed.

Black maternal mortality is a major issue in the United States. In recent years, the maternal mortality rate for Black women has been around three times the rate for White women. According to some research, this may be because people believe Black people are biologically different and can handle pain on a higher level, or because certain healthcare professionals lack empathy. The current study will examine whether participant’s attitudes toward a Black (vs. White) female giving birth will vary according to pain sensitivity, dehumanization, and other moderating variables (e.g., empathy, social dominance orientation). It is hypothesized that participants who read a scenario of a Black female giving birth will report lower pain sensitivity scores and rate her as being less than human as compared to participants who read a scenario about a White female giving birth. The implications of this study in relation to the well-being of Black women will be further discussed.

Visual working memory (VWM) is a critical cognitive capacity for the processing and manipulation of visual information, supporting tasks such as reading, spatial navigation, and object recognition. Cueing, the process of directing attention towards relevant information before (pre-cue) or after (retro-cue) stimulus presentation, has been shown to enhance object recognition and memory accuracy. This study aims to explore the effects of pre-cueing and retro-cueing on VWM capacity in a virtual reality (VR) environment, which offers a more immersive and ecologically valid setting for such investigations. TCU student participants were tasked with comparing a test stimulus to a sample stimulus based on its identity or location, on trials with pre and retro-cues. Based on previous data collected in the TCU Comparative Cognition Lab, it is anticipated that pre-cueing will result in better performance than retro-cueing. Given the increased ecological validity of virtual reality (VR) compared to 2D tasks, this trend is expected to become even more pronounced. The anticipated results of this study could offer valuable perspectives on how visual working memory operates within virtual reality settings and enhance our comprehension of how cueing influences memory retrieval. These findings may have implications for the development of more effective VR-based training and educational programs, as well as for the design of user interfaces that optimize memory recall in immersive environments.

Throughout the United States, industrial agriculture has created a set of traditional methods used to raise beef cattle. These traditional methods have large adverse effects on the environment as well as profitability. The producer who took part in this case study has been managing a beef cattle operation in North Texas since 1999. This rancher’s non-traditional, holistic land management and business approaches to finishing grass-fed cattle for his custom beef brand are drastically different than traditional methods. This single-case study examines the intersection of profitability and sustainability on a traditional beef cattle operation compared to a holistic non-traditional beef cattle operation. The data collected for this study includes interviews, a review of government documents, historical management documents, soil laboratory reports, and botanical and phenological reports. The findings from this study inform land management practices that boost both economic value and long-term environmental sustainability.

There are numerous known chemicals, present in surface waters, which may pose a significant risk to the health of fish living in aquatic environments. These chemicals can impair a variety of physiological process and to date, screening assays for evaluating acute, chronic, reproductive toxicity, and endocrine disruption have been well developed. However, screening assays capable of identify chemicals that can adversely impact immune function have yet to be developed. Developing such assays is necessary given that immune system disruption can increase the incidence of disease and death. Thus, the purpose of this study was to develop and validate a fish-based neutrophil migration assay that can be utilized to rapidly screen chemical for immunotoxicity. The specific objective of this study was to develop a neutrophil migration assay featuring adult fathead minnows (a commonly-used toxicity testing model organism). Specifically, this study sought to optimize methods for two key steps of the neutrophil migration assay – tail injury and depigmentation of fish tails for neutrophil visualization. Three tail injury methods were evaluated including partial amputation, a tail nick, and a biopsy punch. The methods of depigmentation evaluated were H2O2/KOH treatment alone, H2O2/KOH and acetone, and H2O2/KOH treatment in combination with acetone and FlyClear solution 1.1 (Triton X, THEED, and urea). Results showing which of these methods is best suited for neutrophil migration assays featuring adult fathead minnows will be presented.

Oxidative stress is an imbalance of reactive oxygen species (ROS) and antioxidant defenses resulting in cell damage and chronic inflammation. It contributes to many pathologies including neurodegenerative disorders, cardiovascular disease, diabetes, and cancer. Macrophages and microglia are phagocytic immune cells that destroy pathogens while releasing inflammatory mediators, such as pro-inflammatory cytokines and ROS. While inflammation is initially a protective mechanism, chronic inflammation is damaging to tissues. To counter oxidative stress, cells express nuclear factor-erythroid 2-related factor (Nrf2) to mitigate excess ROS production. Nrf2 is a transcription factor that promotes the expression of numerous antioxidant enzymes. Our study targets the expression and activation of Nrf2 in cells treated with L2, a compound created by Dr. Kayla Green (TCU Chemistry). Our lab is attempting to determine the molecular mechanism in which L2 may protect phagocytic cells from oxidative stress, and if this mechanism involves the Nrf2 pathway. This research could provide preliminary evidence for the efficacy of this compound as a treatment option for diseases involving oxidative stress.

With increasing global industrialization and subsequent pollution, there are mounting concerns regarding the presence and impacts of reproductive endocrine disrupting chemicals (REDCs), including environmental estrogens. These concerns have led to new international regulations (i.e. REACH) which require that chemicals be screened for endocrine disrupting activity. A variety of in vivo and in vitro screening currently exist; however, the in vivo methods are time-intensive and expensive and the in vitro methods may fail to detect estrogenic compounds with unique modes of action. Thus, there is a need for in vivo estrogen screen assays that are quick and inexpensive. The objective of this study is to validate the newly-developed Rapid Estrogen ACTivity In Vivo (REACTIV) Assay as a reliable approach for the detection of chemicals with estrogenic activity. This assay employs Japanese Medaka (Oryzias latipes) that have been genetically modified to co-express green fluorescent protein and choriogenin (an egg precursor protein). In the assay, the transgenic medaka embryos are exposed to a chemical of interest for 24 hours after hatch and then imaged under a fluorescent microscope. To validate the performance of the assay, tests were performed using two chemicals with known estrogenic activity (i.e., bisphenol A, estradiol) and two inert chemicals (i.e., saccharin and cefuroxime). Results showed that larvae exposed to the estrogenic compound experienced dose-dependent increases in liver fluorescence, while those exposed to the inert chemicals did not. Overall, these results indicate that the REACTIV assay produces predicable results and thus, may be appropriate for use as a standardized estrogen screen method.

BRCA1 is a gene found in humans that, when mutated, has been linked to breast and ovarian cancer. A homolog version of this gene, known as brc-1, exists in an organism called the Caenorhabditis elegans. This is a species of nematode worm that has the potential to be used as a model organism to study this homolog gene that is associated with human breast cancer. Previous studies with C. elegans have shown links between the brc-1 gene and DNA damage responses, cytochrome p450, or cyp, transcription levels, and ratios of male phenotype worms. This project focused on studying whether these brc-1 functions are dictated by the enzymatic activity of the protein made by this gene. To measure these phenotypes, we used a strain of C. elegans with a brc-1 mutation engineered to lack enzymatic activity of the BRCA1 protein toward nucleosomes. In order to determine how this lack of enzymatic activity affects brc-1 functions, we measured levels of reactive oxygen species (serving as a proxy for DNA damage), numbers of male offspring, and cyp levels in the mutant and wild-type C. elegans. Our initial results indicate the effects of enzymatic activity towards nucleosomes on the aforementioned phenotypes.

Staphylococcus aureus is the causative agent of many skin infections and the leading cause of death due to infectious disease in the United States. Additionally, S. aureus is known to rapidly gain antibiotic resistance, as seen with methicillin resistant Staphylococcus aureus (MRSA). Zinc oxide (ZnO), a nontraditional antibiotic, demonstrates antimicrobial action against S. aureus. While the exact mechanism of ZnO antibacterial action is currently unknown, production of reactive oxygen species (ROS) is a commonly proposed mechanism. We find that S. aureus ΔkatA, a mutant susceptible to hydrogen peroxide (H2O2) due to a deletion in the catalase gene, exhibits comparable growth to wild type S. aureus in ZnO. This suggests that production of H2O2 is not vital to the antimicrobial action of ZnO. To further test this, we generated a ZnO resistant mutant (ZnOR) that demonstrates less susceptibility to ZnO. We find that the ZnOR mutant demonstrates comparable growth to wild type S. aureus in H2O2, making H2O2 production an unlikely toxicity mechanism of ZnO. To evaluate the role of ROS besides H2O2, susceptibility of ZnOR and wild type S. aureus to two other ROS, bleach and paraquat was evaluated. We are currently investigating whether N-Acetyl-Cysteine (NAC), a compound that stimulates production of antioxidants and is protective against a wide range of ROS, protects S. aureus from ZnO mediated toxicity. Our data suggests that ROS formation is not the dominant mechanism of antimicrobial action by ZnO and future studies should focus on other potential mechanisms of action.

Cryptococcus neoformans is an ubiquitous fungal pathogen that is detrimental for immunocompromised patients, leading to pneumonia and fatal meningoencephalitis. Fungal signaling lipids termed eicosanoids have been associated with increased virulence in this pathogen. Since C. neoformans lacks common enzymes associated with eicosanoid biosynthesis in humans, this pathway presents novel genes which could be used as potential drug targets. Our study focuses on EncT, a poorly characterized gene which encodes an efflux pump and is involved in the production of eicosanoids in C. neoformans. To evaluate the potential role of this gene in virulence, we used CRISPR technology to knock out (KO) the EncT gene, followed by screening for stable mutants and confirming the gene deletion via DNA amplification. After constructing the KO, we conducted in-vitro virulence assays of the KO strain and the wild-type strain (H99), including tests to assess sensitivity to temperature and changes in virulence factors including the production of melanin and capsule. These tests will help us characterize the potential role of the EncT gene in the virulence of this pathogen. Future directions include using a similar gene-editing method to generate an EncT reconstituted strain for use as an additional control in the in-vitro assays. Further, H99, the KO strain, and the reconstituted strain will be given to mice to evaluate the pathogenicity of the KO strain in an in-vivo model. Additionally, we will evaluate the presence of the fungi in the lung and the dissemination in other organs, and we will analyze the host immune response. By knocking out a gene involved in virulence-associated lipid production and characterizing the role of this gene in pathogenicity, this project will broaden the knowledge of the role of lipids in fungal pathogenesis and provide information that could potentially assist in developing therapies against this pathogen.

In a remarkable work of symbiosis, the gut microbiota coordinate with the brain to regulate multiple bodily functions, including those of the immune system, through bidirectional communication with the gut-brain axis. This symbiotic process has been shown to affect human health and disease pathology as certain inflammatory responses correlate with the composition and general disruption of the gut microbiome. To name a few, neurological disorders, gut-based inflammatory disorders, and cancer have been linked, in part, to dysfunction of the gut-brain axis. Previous literature on the gut-brain axis stems from in vivo and in vitro models, which have worked to understand the connection between the microbiome and disease pathology. Emerging evidence from these studies has continued to become more convincing regarding the importance of the bidirectional relationship in human health. In this review, evidence focusing on the intricate connections between the gut-brain axis and several inflammatory diseases, including irritable bowel syndrome, celiac disease, Crohn's disease, cancer, lupus, and Alzheimer’s disease, will be discussed. How this information can be utilized, including what has been or could be done in the clinic to improve the outcomes of patients with inflammatory-related diseases, will be highlighted so that continued advances in this newer aspect of medicine might lead to direct benefits for human health.

Bacillus anthracis is a gram-positive bacterial pathogen that causes the deadly infectious disease anthrax. B. anthracis contains over 5,000 chromosomal genes, and we believe there are unidentified chromosomal genes important for virulence. Our lab constructed a transposon mutant library with random disruptions in the B. anthracis Sterne genome to screen for novel virulence factors, and we have previously identified two virulence genes, clpX and yceGH, using this library. In this screen, we used hydrogen peroxide, a reactive oxygen species involved in innate immune defense, and screened around 1000 mutants. We obtained three mutants that were susceptible to hydrogen peroxide in vitro: 11F11, LV1, and LV2. To determine whether they also had phenotypes in vivo, we infected Galleria mellonella to study their virulence in an invertebrate animal infection model. LV2 showed reduced virulence in the in vivo survival assay, and all three mutants showed reduced virulence in the in vivo competition assay. I have determined the site of the transposon insertion in 11F11 and LV1, and the transposon has inserted in the genes for catalase and a collagenase-like protein, respectively. I am currently creating an independent insertional mutation in LV1 to confirm that the observed phenotypes are linked to the disruption of the collagenase-like protein. Future directions include creating a complementation plasmid for LV1 and determining the insertion site of LV2. The findings of this research could be used as potential therapeutic drug targets and will offer insight into the mechanisms that B. anthracis uses for its pathogenesis.

Alzheimer's Disease (AD) affects approximately 6.5 million Americans, and there is currently no cure. Prior research has shown that a key pathology of AD is amyloid beta, a protein that aggregates and form plaques in the brain, under pathological conditions. If amyloid beta is not cleared by the body, resultant plaques may disrupt proper cognitive and neuronal function. As the liver plays a crucial role clearing amyloid beta, liver damage may jeopardize the efficacy of the liver to clear amyloid beta in the periphery of the body, enabling it to reach the brain.

One way liver function can be disrupted is through diet, specifically the Western diet (WD), which has been shown to cause non-alcoholic fatty liver disease (NAFLD) and inflammation, both of which are associated with AD. A WD is classified as one that contains high amounts of refined sugars and saturated fats derived from animals. Conversely, the Mediterranean Diet (MD), a largely plant-based diet, contains high amounts of monounsaturated fatty acids and polyunsaturated fatty acids. These dietary factors have been shown to decrease inflammation and increase antioxidant effects, further protecting the brain from AD pathology. Therefore, we hypothesize that the MD could protect the liver and be used as a potential prevention strategy for NAFLD and AD.

The current study examined the effects of WD and MD on the relationship between the liver and the brain in wild type mice. During tissue collection, livers were taken and histologically analyzed. The livers from each experimental group were processed, stained, and evaluated for their overall composition.

BRCA1 and BARD1 are proteins involved in the repression of genes associated with increased risk for breast and ovarian cancers. This is accomplished through ubiquitination of H2A and subsequent changes in chromatin compaction. BRCA1 and BARD1 form an E3 ligase (BCBD complex), and mutations affecting the enzymatic functions of this complex can predispose women to these cancers. The model organism C. elegans contains orthologs of these proteins, BRC-1 and BRD-1, which makes it a useful organism for studies of protein function; however, little is known about the mechanism of ubiquitination in C. elegans as compared to humans. This project used nucleosome assays to provide more insight on the ubiquitination of H2A by the BCBD complex in C. elegans. The objectives of this project included characterizing the interaction of the BCBD complex with H2A and identifying a specific lysine target in C. elegans. The conserved lysine targets were mutated out of H2A and nucleosome assays were performed to identify potential reductions in ubiquitination activity. In addition, we hypothesized that enzyme-substrate interactions, specifically between H2A and BRD-1 in C. elegans, are important in directing ubiquitin to the target site. Amino acid residues in BRD-1 thought to be important for these interactions were mutated out, and assays were performed to assess changes in ubiquitination activity. The H2A nucleosome assays showed that the mutations of conserved lysines in the H2A N-terminus and C-terminus in C. elegans did not significantly reduce ubiquitination activity, and a definitive target could not be identified. However, the BRD-1 assays identified amino acid residues in C. elegans that participate in directing the ubiquitination process. Further studies are needed to determine if C. elegans has any preferential lysine targets at a non-conserved residue or if it is truly nonspecific in its activity. Currently, mass spectrometry analysis is being performed as a complementary method to attempt to pinpoint the location of lysine ubiquitination.

Alzheimer’s Disease is a neurodegenerative disease characterized by cognitive, functional, and neuronal loss. Its core pathology includes beta-amyloid protein plaque formation, neurofibrillary tangles of tau protein, and loss of microglial cell function, all of which may be facilitated or exacerbated by a prolonged neuroinflammatory response. The inflammatory signaling pathway culminates in the activation of transcription factor NF-κB, which then goes on to activate the expression of cytokines and other signaling molecules such as TNFα. One of the points of regulation for this pathway is the constitutive binding of the IκBα protein to NF-κB that prevents NF-κB from entering the nucleus. However, when the appropriate stimulus triggers the pathway, a downstream effect is the phosphorylation of IκBα by the IκB kinase, and its subsequent degradation which then releases NF-κB for translocation into the nucleus.
This project aims to elucidate the mechanism of action of novel anti-inflammatory drugs (provided by P2D Biosciences company). Previous in vivo studies with the compound have shown a reduction in inflammation and improved cognition, but the drug’s exact point of interference in the pathway remains unclear. Therefore, this project aims to assess if the drug reduces inflammation by reducing IκBα degradation, thus preventing NF-κB from being able to turn on cytokine expression.
BV-2 mouse microglial cells were exposed to the drugs, followed by exposure to LPS for various time intervals, then harvested and lysed. A Western blot procedure was performed on the lysates to visualize the amount of IκBα present, then those bands were quantified to compare against control cells that were not incubated with the drug. It follows then, that if the drugs’ mechanism of action is inhibition of NF-κB release into the nucleus, then there will be increased amounts of IκBα in the treatment cells compared to the control cells as IκBα degradation is prevented.