Post-Traumatic Stress Disorder (PTSD) is one of the most common mental disorders in the world. People with PTSD often have difficulty managing time or have witnessed a traumatic event in the past. PTSD is also associated with memory issues which are characterized by intrusive memories that can cause re-experience aspect of the traumatic event. Through the use of computational models, we aim to investigate the differences in memory retention in spaced vs massed practice presentations. We hypothesize that mass practice will lead to less accuracy in recognition and less intrusive thoughts during retention. Through recruiting healthy adult participants with no history of PTSD or other stress-related psychiatric disorders we get a base data avoiding discrepancies. Participants view visual stimuli as images, either neutral or triggering which are presented in pseudo-order and are asked to rate them based on their valence ranging from neutral to extremely negative. The images are presented under two conditions: mass and space presentation. For spaced presentation, different versions of the triggering image category are presented in no particular order with neutral and filler images in between. For massed presentation, all versions of the triggering image category are presented one after the other with no neutral or filler images in between. 24 hours after the initial presentation of the images, participants are tested on memory retention in the form of image recognition. Participants are asked about the difficulty of recalling the image and how often they think about the image. Having collected data during AUT 24, we were able to understand there is a correlation present between memory retention in mass and spaced recognition in terms of traumatic and non-traumatic events. By collecting data through WIN 25, we will have greater accuracy in terms of significant data.

Alzheimer’s disease (AD) is a progressive neurodegenerative disease that affects millions of people. Repetitive transcranial magnetic stimulation (rTMS) is a noninvasive stimulation typically used in psychiatric conditions such as depression and anxiety. rTMS works by using an electric current to generate a transient magnetic field, depolarizing neurons in a target region and creating lasting changes in brain connectivity via synaptic plasticity. Patients with AD show disruptions in the Default Mode Network (DMN), a network of brain regions typically active during rest and crucial for memory consolidation. We hypothesize that strengthening the DMN through rTMS targeted at the left Brodmann 8AV region, selected for being an easily accessible node of the DMN, will improve memory in AD patients. To test this hypothesis, we are conducting a single-blind, single-arm, randomized cross-over trial of rTMS on early-stage AD patients over a 12 week period with week 1 where we scan for the 8AV region via MRI, during week 3 and 8 being the placebo or treatment week. We measure our primary outcome of the participants’ speed of forgetting —a novel index of memory function—through an individualized, adaptive memory test. To eliminate potential confounding variables, we also measure depression and anxiety symptoms during the 1st, 8th and 12th week of the study. Additionally, functional MRI scans will be analyzed for potential structural or functional differences caused by treatment. Preliminary results from our initial participants have shown promising improvements, and we are hopeful that similar outcomes will be observed in the remaining participants. Successful results would provide a novel target for AD treatment using rTMS, and support further investigation of rTMS as a viable treatment option.

Changing patterns of human land use near coastal zones have increased the abundance of aquatic habitats where mosquito and midge larvae develop. Salt-tolerant species, such as Aedes togoi mosquitoes and Dicrotendipes enteromorphae midges, have evolved distinct anatomical and physiological mechanisms that allow them to thrive in saltwater environments while also being able to develop in freshwater. While much is known about how freshwater-confined species maintain salt and water homeostasis (i.e. osmoregulation) in dilute freshwater, these processes remain largely unexplored in salt-tolerant species. The main goal of this research was to describe and compare the osmoregulatory strategies of Ae. togoi and D. enteromorphae larvae reared in seawater and freshwater, focusing on how they maintain ion and water balance in saline conditions. Based on information from other salt-tolerant insects, I hypothesized that the rectum of these species is the main organ that excretes salts and will have higher expression of ion pumps compared to other osmoregulatory organs of the larvae. This increased expression would support the secretion of hyperosmotic (salty) urine, a critical adaptation for survival in saltwater. Using immunohistochemistry and fluorescence microscopy, I localized key ion-transport pumps within the rectum and other osmoregulatory organs of Ae. togoi and D. enteromorphae larvae. I identified a high expression of Na⁺/K⁺-ATPase (sodium/potassium pump), and V-type H⁺-ATPase (proton pump) enriched in the anal gills, renal tubules, and the rectum. My findings suggest that the coordinated function of many osmoregulatory organs, and not just the rectum, allows for salt-secretion by marine insects, and this establishes an initial framework of cellular mechanisms among marine insects. This research has implications for predicting future species distributions with rising sea levels, as well as the potential to identify novel targets to control salt-tolerant mosquito populations to mitigate disease transmission in coastal regions.

Neotrypaea californiensis, or burrowing shrimp, is a native pest for oyster growers in Washington. The shrimp create networks of interconnected burrows that displace and liquify sediment, suffocating oysters. The pesticide carbaryl was used for decades to effectively control shrimp populations. More recently, the Environmental Protection Agency (EPA) deemed carbaryl an environmental hazard, leading to a vital need for a shrimp control method with minimal non-target effects. Very little is known of the physiology of shrimp that allows them to tolerate environmental extremes. My study aims to understand the tolerance of Netotrypaea californiensis to high concentrations of salts that challenge their ability to regulate internal ions, pH, and metabolic wastes. Using a laboratory-based system whereby shrimp burrow in 6 inch deep sediment in seawater, my research findings revealed that the addition of high concentrations of sodium bicarbonate (NaHCO₃) caused shrimp mortality while high concentrations of regular sea salts did not. This suggested that an ionic imbalance—high levels of sodium and reduced levels of chloride—may be causing shrimp death. To investigate, I collected the gills, a critical organ that regulates systemic ion levels, of shrimp after NaHCO₃ exposure and quantified the abundance and localization of key ion pumps using Western blotting, immunohistochemistry and confocal microscopy. I found a significantly higher expression of Na⁺/K⁺ pumps in the gills with NaHCO₃ exposure, suggesting a major ion-regulatory disturbance caused by NaHCO₃. These data will expand our understanding of how salts that cause ionic imbalance in seawater can disrupt the internal levels of ions that are critical for most biological processes. Findings will be disseminated to Washington oyster growers to aid in the management of their crop.

The external fertilization and transparent embryos of zebrafish make them an informative model of vertebrate embryonic development from the 1-cell stage. In this study, we examine the impact of de novo GTP synthesis on the formation of the embryonic somites, which are embryonic cells which develop into segmented blocks of muscle that run the length of the body. We hypothesize the de novo GTP synthesis is required for the correct patterning of somite borders in zebrafish embryos, and that this process facilitates the formation of a vertebrate body plan. Inosine monophosphate dehydrogenase 2 (IMPDH2) is the enzyme which catalyzes the conversion of inosine monophosphate (IMP) towards the de novo synthesis of GTP instead of ATP. To test the impact of de novo GTP synthesis on somite formation, we inhibited IMPDH2 function with mycophenolic acid (MPA) both before and after somite formation began. MPA caused stronger defects in the somite morphology and embryonic body shape when added to embryos before somite formation began, earlier in development. We performed in situ hybridization against xirp2a to assess the effect of inhibiting IMPDH2 function on the formation and patterning of the somite borders. MPA treatment decreased the definition of somite borders we could observe in the posterior tail. Inhibiting IMPDH2 with MPA produced somites with smooth, round borders instead of the chevron-shape typical of zebrafish. We next conducted immunohistochemistry against IMPDH2 to examine the expression and localization of this enzyme in embryonic cells when GTP conditions are low. In MPA-treated embryos, we observed increased expression of IMPDH2 across the entire embryo. We will next explore how GTP abundance affects activity of the clock, a mechanism which synchronizes gene expression of embryonic cells.

Hyalella azteca are freshwater amphipod crustaceans abundantly found throughout North America and are frequently used in toxicology for water and sediment toxicity testing. As freshwater bodies change due to anthropogenic climate change, understanding chronic, sublethal impacts to aquatic life is critical. Amphipods are known to have a higher tolerance to heavy metals (i.e. Cu²⁺) and road salt (i.e. NaCl) compared to other aquatic invertebrates, however, these contaminants may have subtle, sublethal consequences on their ability to smell and detect chemical cues for survival and reproduction. This research identifies the combined impacts of environmentally relevant levels of Cu2+ and NaCl, on the olfactory system and olfactory-related behaviors of H. azteca. H. azteca underwent acute 96 hour exposure to control, Cu2+ (30 µmol/L), and combined Cu2+ (30 µmol/L) and NaCl (5 ppt) contaminated freshwater. I recorded daily measurements of survival, and utilized qPCR to examine changes in the expression of key olfactory genes that we predicted would be modulated in response to these multiple stressors. I assessed the olfactory-associated foraging behavior to determine changes in their detection of an attractive food cue when exposed to Cu2+ and NaCl, which would indicate olfactory impairment. Preliminary results show that amphipod survival is not impacted by Cu2+ and NaCl contamination but changes to their olfactory system occur. Therefore, U.S. Environmental Protection Agency water quality standards for heavy metals and salt may not mitigate long-term, sublethal effects on aquatic animal populations as it relates to this important sensory modality.

Certain species exhibit the remarkable ability to regenerate their appendages, a process that requires complex metabolic pathways to facilitate the cellular proliferation needed to regrow tissue. Among these species, Xenopus tropicalis, the Tropical Clawed Frog, serves as a great model for regeneration studies because of its transient regenerative capacity. X. tropicalis tadpoles exhibit the ability to regenerate their tails, but this capability is gradually lost after metamorphosis. This unique characteristic allows for direct comparison of regenerative and non-regenerative processes within the same species. Previous work from the Wills lab indicates that genes encoding the enzymes of the pentose phosphate pathway (PPP), which generates precursors of biosynthetic molecules such as nucleotides and lipids, are highly expressed during tadpole tail regeneration. Although tail regeneration has been well studied, the variation in hind limb regenerative capacity across developmental stages and the metabolic pathways involved remains unclear. Hence, I performed a live imaging study to determine the developmental progression of hind limbs and assess their regenerative potential. This data suggests a decrease in success as the tadpole gets older. Immunohistochemistry staining of mitotic cells in developing limbs shows that proliferating cells decline as regeneration competency decreases. I hypothesized that genes for the PPP enzymes would also be expressed during successful limb regeneration, which was confirmed by in-situ hybridization. Together, these findings indicate that the regenerative capacity in limbs of X. tropicalis is stage-dependent and that PPP genes are expressed during all stages of regenerative capacity. This provides insights into the role of metabolic reprogramming in appendage regeneration, with the potential for translating it into non-regenerative species like mammals.

Autism is a neurodevelopmental condition characterized by social-communication differences and restricted, repetitive behaviors (American Psychological Association, 2013). While racial disparities in autism prevalence have been documented, not much research has examined how these differences appear in autism screening measures. This study investigates racial differences in Autism Spectrum Quotient (AQ) scores among non-autistic Asian and White adults to assess potential biases in autism screening. Using data from three NIH-funded studies (COBRA, BEAM, and The Korean Study), AQ scores from 166 participants (n=102 White, n=64 Asian) will be analyzed through t-tests and Repeated Measures Analysis of Variance (ANOVA) to compare total and subscale scores. Preliminary observation suggests that non-autistic Asian participants may be more likely to score higher on the AQ, raising concerns about false positives in autism screenings. Understanding these differences will help improve diagnostic accuracy, reduce disparities, and promote equitable access to neurodevelopmental resources.

Overgeneral Autobiographical Memory (OGM) is a common symptom of depression and Post-Traumatic Stress Disorder (PTSD). Instead of remembering specific details, individuals with OGM can only describe past events from their lives in general terms. The "trauma hypothesis" suggests that OGM emerges because individuals suffering from PTSD tend to reduce the number of details they retrieve about their memories to avoid remembering their trauma. However, this hypothesis does not fully explain how this avoidance is learned, or why avoidance spreads from traumatic memories to all autobiographical memories. To this end, we propose a computational model of OGM that integrates theories of memory retrieval and trauma with reinforcement learning. In this model, multiple episodic memories are nodes in an interconnected network, and memories are retrieved when visiting that node in the network. The more nodes that are visited, the more detailed that autobiographical recall will be. On the other hand, visiting more nodes comes with an increased risk of encountering a traumatic memory, which comes with negative emotional valence. The decisions about whether to visit another node or terminate the retrieval process are made using reinforcement learning, which takes actions based on predicted outcomes. By obtaining a greater understanding of OGM through this model, we hope to improve treatments for PTSD that specifically targets its effects on memory. 

Proteins in the NFкB family are transcription factors that modulate the expression of genes relating to immunity and inflammation. One protein within this family is the p50/RelA heterodimer which includes a structured DNA-binding domain and a Transcription Activation Domain (TAD) which is intrinsically disordered, or naturally lacking secondary and tertiary structure. Previous studies have shown that interactions between the DNA-binding domain and TAD affect DNA binding affinity and specificity. With the goal of further assessing and comparing the interactions between intrinsically disordered and structured domains in proteins of this family, we have worked to troubleshoot and optimize protocols relating to expressing and purifying human p50/RelA protein. I analyzed the efficiency of multiple chromatography steps and introduced changes to improve yield. Optimization of the expression and purification protocols will enable future investigations into the DNA binding activity of this protein.

While taking multiple general science courses and dance courses as a double-degree student, I have devised creative ways to balance my studying and dance training. One involved creating dance choreography to memorize organic chemistry reactions, which inspired me to choreograph a dance piece named after the motor cortex “Homunculus” for Aura Dance Company’s (RSO) annual spring show in 2023. This sparked my interest in organizing a project to teach this learning structure that may be useful to others. With help from Professor Hannah Wiley and MFA candidate Beth Twigs, I designed dance workshops for my peers to learn more about neuroscience and dance. NeuroDance is a multidimensional project to educate students about neuroscience through dance-making tools. The project involved organizing workshops where participants learned movements inspired by molecular neuroscience, neuroanatomy, and skeletal anatomy. Participants modeled ions, neurons, and planes of movement through facilitated movement phrases and seeds. To assess learning outcomes, quizzes were given before and after the workshops. Volunteers were recruited from on-campus social, dance, and neuroscience groups, and outreach will occur via social media and posters. The data from the learning aspect of these workshops house the scientific results, but the movement observed served as the foundation for a larger choreographic work presented in the Department of Dance’s Dance Majors Concert (2025). The physicality and repetition inherent in dance offer a unique and enriching platform for learning. I aim further to explore the potential of dance education in STEM with this pilot study.

Autism is a neurodevelopmental disorder characterized by differences in social-communication and the presence of restricted and repetitive behaviors and interests. Many autistic individuals engage in “camouflaging” to hide or change their behaviors associated with autism to avoid social stigma. This study aims to explore neurophysiological characteristics underlying camouflaging. Electroencephalography (EEG) is a popular psychophysiological tool that measures brain activity through oscillatory patterns, reflecting various cognitive and emotional processes. Specifically, during “resting state” (when the brain is exposed to minimal external stimuli), theta waves have been shown to have increased activity during periods of increased cognitive load, attentional demands, and task difficulty – mental states that all relate to camouflaging based on qualitative research.  Participants included autistic (n=108) as well as non-autistic adults (n=85), between the ages of 15 and 31 years. Participants completed the Camouflaging Autistic Traits Questionnaire (CAT-Q) which measured three domains of camouflaging in autism: compensation, masking, and assimilation. EEG recordings were taken during resting state and oscillatory activity in the theta frequency band (4-8 Hz) will be analyzed. Our hypothesis is that camouflaging traits will be positively correlated with theta wave activity.  Camouflaging can lead to various challenges for autistic individuals, including depression and anxiety. Thus, identifying the proposed analyses could provide valuable insight into the cognitive and emotional processes of camouflaging, ultimately contributing to a better understanding and potential treatment for mental health challenges faced by the autistic community.

Gray whales in the North Pacific annually migrate north to the Gulf of Alaska and the Bering Sea, and their migration route bypasses the Salish Sea. Roughly a dozen of these whales, commonly called “the Sounders,” have detoured their migration into North Puget Sound since the 1990s. These whales have been observed feeding on ghost shrimp in the intertidal area of sediment beaches in North Puget Sound, using a high risk strategy of feeding on shrimp at high tides. This feeding strategy leaves large indents, or “feeding pits”, in the sediment that are revealed at low tide and can provide insight into the Sounders’ feeding habits and contribute to a deeper understanding of the North Pacific gray whale population. My research focused on locational trends of gray whale feeding pits on Jetty Island West beach, and I observed longitudinal locations of specific pits in the intertidal zone to investigate feeding patterns. I observed feeding pits with drone imagery collected at low tide and compiled into aerial maps, or “orthomosaics,” and I compared feeding pits in different longitudes to observe where on the beach whales are feeding. Two seasons of feeding pit imagery were collected from late winter and spring of 2024 and 2025, and I have analyzed the imagery using ArcGIS pro. Survey site area ranged from approximately 0.09km² to 0.4 km² for different maps. The non-invasive nature of drone photogrammetry has recently increased its use in marine and biological research, and this method of data collection is ideal for surveying gray whale pits on Jetty Island. Because of the increased risk of feeding in higher tidal zones, I expect to find higher concentrations of feeding pits at lower tidal zones.

Seabirds are considered a strong indicator species for ecosystem health due to their visibility, lack of behavioral and phenotypic plasticity, and high trophic level.  Current declines in seabird populations are often attributed to bottom-up ecosystem control regulating upper trophic level populations. These bottom-up effects might be caused by reductions in marine productivity due to climate change. I performed statistical and graphical analyses on the National Audubon Society’s Christmas Bird Count data from Puget Sound and water chemistry data from the Ocean Research College Academy’s moored and deployable sensors. This allowed me to identify possible relationships between bird populations and water chemistry from 2009 to 2024 in the Possession Sound estuary. My initial analyses demonstrated the expected decline in collective seabirds counted, however certain pelagic species experienced unexpected increases. Further investigation is required to determine whether the increase was caused by ecosystem dynamics or improved count methods. My initial analyses did not indicate any relationship between water chemistry and bird populations. The lack of apparent relationship may be due to the water chemistry changes having impacts on primary productivity and indirect bottom-up trophic cascades, which could have a significant lag time in effects on bird populations. My analysis also does not account for environmental factors in disparate migration sites or breeding colonies that might affect bird populations. 

Autism spectrum disorder (ASD) and attention-deficit/hyperactivity disorder (ADHD) are neurodevelopmental conditions that are known to co-occur with decreased sleep quality and quantity. ASD is characterized by social communication differences and restricted, repetitive behaviors and interests.  ADHD is characterized by inattention and/or hyperactivity/impulsivity. Previous research found higher instances of sleep disruptions within adults with ADHD as well as autistic populations compared to neurotypical adults. However, quantitative data around the associations between ASD and sleep disruptions is sparse.  This leads to our research question which is, how does sleep disruption affect autistic, autistic + ADHD adult’s sleep quality and quantity compared to neurotypical adults? We hypothesize that ASD+ADHD and ASD adults have experience higher sleep disruptions (fewer hours of sleep and lower quality) compared to non-autistic adults. Participants included autistic adults (n=66), autistic adults with ADHD (n=39), and non-autistic adults (n=221) enrolled in the NIH-funded COBRA and BEAM studies, which investigated how the brain processes sensory and visual information.  Autism diagnoses were confirmed using standard assessments (such as Autism Diagnostic Observation Schedule, 2nd Edition, Autism Quotient) by clinicians.  Participants self-reported ADHD diagnoses, sleep quantity (hours per night on weekdays, hours per night on weekends), and sleep quality (over the past week on a 4-point Likert scale). An ANOVA will be used to compare sleep quality and quantity between diagnostic groups. Understanding neurodivergent populations’ sleep quality and quantity helps improve public health communication around sleep health, which is particularly important population health issue given the link between sleep, mental and physical health. Finally, identifying populations most in need of sleep interventions helps us address the increased number of adults with sleep disturbance and disorders.

Patient-provider communication impacts healthcare outcomes, but assessing the quality of interactions manually takes time and effort. This project explores the automatic assessment of patient-provider interactions using Language Style Matching (LSM). LSM scores the linguistic similarity of function words between conversational partners (e.g.,  pronouns, articles) from 0 (low matching) to 1 (perfect matching), reflecting how in-sync partners are. Past research establishes LSM as a marker for the quality of interpersonal communication that predicts how likely romantic relationships are to last, but has not been explored for clinical interactions. We (CL, NE) applied LSM to investigate how well patients and providers matched each other's speaking styles for insights into the quality of clinical interactions. We used Linguistic Inquiry and Word Count (LIWC), a software program for LSM analysis of transcripts. Using LIWC, we analyzed the transcripts of 108 simulated visits between 54 primary care providers and four standardized patients. We used descriptive statistics to characterize LSM across visits. Our initial findings show that LSM scores range from 0.77 to 0.94 ( mean=0.86, SD=0.03) which is similar to prior research where most verbal conversations fall between 0.83 and 0.94. These findings show that on average providers and patients tend to match each other in their speaking style at a level similar to typical conversations. However, we identified some outliers that fall below 0.83 threshold. Opportunities for future work include thin-slice analysis of the transcripts to understand how LSM scores change throughout a visit and comparing LSM scores to self-reported survey data about visit quality. ​​We hope to further investigate this efficient marker of conversational quality as LSM has the potential to characterize the quality of clinical interactions without the time and effort required of traditional manual approaches.

The necessity of criminal justice reform has grown increasingly significant as governments address the challenges of mass incarceration and its far-reaching social and economic consequences. Over the last decade, California has been spearheading reform in the United States. In compliance with a Supreme Court ruling, California passed Proposition 47 (Prop 47)—a landmark policy that reclassified certain nonviolent felony offenses as misdemeanors—to reduce the population of nonviolent offenders in the prison system. While Prop 47 successfully reduced incarceration rates and state expenditures on corrections, critics argue that it has also contributed to increased property crime, particularly retail theft, due to theft-felony threshold being increased from $400 to $950. Utilizing a difference-in-differences methodology, this study compares retail business activity in California with a synthetic control group composed of demographically and economically similar states with felony thresholds similar to California’s, pre-Prop 47. The analysis draws on data from the U.S. Census Bureau, incorporating crime rates, business permit activity, and economic trends. This study aims to assess whether Prop 47 led to a tangible increase in property crime and, in turn, a decline in retail business sustainability. The findings will provide empirical insights for policymakers seeking to balance criminal justice objectives with economic stability, informing on the broader implications of sentencing reforms on local economies.

Chronic pain is a public health crisis that has been clinically demonstrated to disrupt reward learning and motivation in affected individuals. Previous literature has indicated that Calca neurons in the parabrachial nucleus (PBN) play a key role in the sensory and emotional processing of pain and become hyperactive in chronic pain models. Despite this, how PBN Calca signalling impacts adaptive decision-making in a positive-reinforcement context remains unclear. This study aims to explore how chronic PBN Calca hyperactivity impacts learning and motivation. Using chemogenetics, a technique that selectively modulates neuronal activity, we chronically activated PBN Calca neurons in transgenic mice. These mice were then tested in a two-phase positive-reinforcement operant conditioning paradigm to assess how chronic PBN Calca activation altered learning rates and motivation compared to controlled animals. In phase one, mice underwent a fixed ratio schedule in which they learned to press a lever during a distinct cue to obtain a food reward. In phase two, mice underwent a progressive ratio schedule in which they had to press a lever an increasing number of times to obtain a food reward. We hypothesized that chronic activation of PBN Calca neurons would impair both learning rate and motivation. With this work, we hope to clarify the impact of centrally-mediated chronic pain on motivational and cognitive processes, which could inform the development of future therapeutic strategies.

This study investigates the Namonuito region of Micronesia to determine whether the many guyots and atolls in this region contain specific slope conditions suitable for cobalt crust growth. This project also investigates whether there is a relationship between suitable zones and general bathymetry. The primary research focused on NAM-2 Atoll and Enterprise, Essex, and Namonuito Guyots. After collecting multibeam and sub-bottom data, the slopes were analyzed for slope angle, optimal slope distribution and composition. Then the results were compared with each other to determine if there are any similarities or correlations. Among all the guyots it was found that almost all the optimal zones were found in gullies, ridge slopes, and at the bottom of areas that experience slope failures. However, there isn’t a clear correlation between mean slope angle, optimal slope distribution and slope face orientation. In addition, sub-bottom data showed that almost all the guyots northern slopes were covered by a single thick pelagic layer, while the southern slopes of Namonuito and Enterprise contained thin pelagic layers with zones of exposed hard substrate. NAM-2 Atoll also was entirely covered in a single thick pelagic layer. Overall, by conducting a geomorphology and sub-bottom comparison test it is possible to narrow down locations of interest that can be further surveyed. The implications of cobalt crust research are that cobalt crusts are another natural source of cobalt which is becoming increasingly difficult to obtain on land. If the Namonuito region contains a large cobalt crust and the cobalt crusts can be extracted, it could have a huge impact of global technology and manufacturing industries.

Humans have limited regenerative capabilities, providing incentive to study other natural models of regeneration to make advances in the field of regenerative medicine. In response to injury, species including Xenopus tropicalis employ cellular mechanisms to replenish lost tissue, a process that has high metabolic demands. Depending on their developmental stage, X. tropicalis tadpoles exhibit different regenerative capabilities after tail amputation, posing them as a unique model system. Three-day-old tadpoles (NF stage 41) are able to regenerate their tails completely after injury, but transiently lose this ability during what is known as the refractory period. However, they soon regain regenerative capabilities in the tail and in the developing hind limb before permanently losing them during metamorphosis. Previous work by the Wills lab has determined that the pentose phosphate pathway (PPP) is required for successful tail regeneration in stage 41 tadpoles, but leaves open the question of whether the PPP remains significant at subsequent regenerative stages and structures. Here I test the hypothesis that the PPP continues to facilitate appendage regeneration in post-refractory tadpoles. To functionally test the requirement of the PPP in post-refractory tail regeneration, I performed pharmacological inhibition of g6pd, a key enzyme in the PPP, during tail regeneration. To assess regeneration quality, I developed a pipeline using FIJI ImageJ and R to quantify metrics of regenerative success such as tail area and length. Using this framework, I found that post-refractory tadpoles had diminished regenerative success under PPP inhibition similar to stage 41 tadpoles. These results suggest that the PPP is required at all stages of tadpole tail regeneration and will provide a more comprehensive understanding of metabolism during regeneration, a potentially beneficial insight for research in wound-healing initiatives in mammals. 

The NF-KB family of transcription factors regulates genes involved in immunity, inflammation, and other biological processes. Members of the NF-KB family can form homo- or heterodimers, which contribute to specific responses to various stimuli. The p50/c Rel heterodimer, an important player in adaptive immunity, regulates gene expression, but its DNA-binding specificity and regulatory mechanisms remain incompletely understood. This study investigates the expression and purification of recombinant p50/c Rel heterodimers. I expressed recombinant c Rel and p50 in Escherichia coli and purified the proteins using Ni²⁺ affinity chromatography. SDS-PAGE analysis confirmed the successful isolation of both proteins at the expected molecular weights. This work lays the foundation for further biochemical characterization, including the investigation of their DNA-binding properties and role in immune signaling. These findings contribute to the understanding of the p50/c Rel heterodimer's function in NF-KB mediated gene regulation. Future studies are needed to explore its DNA-binding specificity and how these interactions impact immune responses and diseases such as cancer and inflammatory disorders. 

The p50/RelA dimer is an essential part of the NF-ĸB signaling pathway, which is responsible for regulating inflammation and immune responses. Most prior biochemical research focused on the mouse version of the p50/RelA dimer. While the findings are useful, its implication to human health remains unclear. This raises the question, how effective do experiments involving mouse proteins reflect those involving humans? We used protocols to express and purify human and mouse p50/RelA dimers, aiming to generate proteins for structural and functional analysis. In the first stage, recombinant protein expression and affinity chromatography techniques were used for purification of both proteins, followed by an SDS-PAGE to assess molecular weight and stability. We found that mouse proteins showed higher intensity bands compared to human proteins, indicating a higher yield. This suggests stability factors as well as potential differences in degradation rates between species. In the second stage, ion exchange and size exclusion chromatography were used to further purify the proteins. During ion exchange chromatography, neither protein bounded as effectively as expected, highlighting the need for protocol optimization. Improving the chromatography conditions will help increase stability and yield of both proteins allowing for more accurate comparisons between the mouse and human p50/RelA dimers. These optimizations are important because it will improve our ability to compare NF-ĸB pathway functionality between species and ultimately make it easier to translate findings from mouse models to human health.