mTOR, the mechanistic target of rapamycin, is a serine/threonine kinase which is an enzyme that phosphorylates the hydroxyl group on a serine or threonine side chain. mTOR kinase regulates protein synthesis, cell growth, and metabolism in response to nutrients and energy in most eukaryotes. mTOR consists of two distinct complexes, mTORC1 and mTORC2. These complexes can be further divided into three components: mTORC1 consists of mTOR, Raptor, and mLST8, and mTORC2 consists of mTOR, Rictor, and mLST8. mTORC1 is critical in metazoan development and has been implicated in aging, cancer, diabetes, cardiovascular disease, and hypoxia, a state in which tissues are deprived of oxygen and can not carry out normal metabolic activity. Previously, the Crowder lab conducted a mutant screen in C. elegans, for hypoxia resistant mutations, and has recently identified a missense (changes one amino acid to another) reduction of function mutation in the daf-15 gene, the C.elegan ortholog of Raptor. A unique feature of this mutation is that the function of Raptor can be turned on and off by varying temperature. It has normal hypoxia resistance at 20 degrees, increased resistance between 21-22, and developmentally arrests at 25 degrees. I and the other authors made use of this conditional developmental arrest phenotype to screen for genetic suppressors, mutations that suppressed developmental arrest. Using genetic mapping, sequencing, and complementation testing, we have identified multiple mutations in three different genes responsible for restoring Raptor function. Preliminary results show mutations in the gene rnf-126 (ring finger protein) suppress the Raptor mutation. Current work by myself and others is designed to answer how these genes control Raptor function and hypoxia sensitivity. Elaborating the function of these genes will define novel mechanisms whereby Raptor and mTORC1 controls metabolism, hypoxic injury, and development.

Mutations in β-myosin heavy chain (MHC) have been implicated in the manifestation of cardiomyopathies. One such iteration of disease, dilated cardiomyopathy (DCM), results in a progressive loss of muscle in the heart, causing a dilation of the chambers that makes it more difficult for the heart to pump blood and leads to progressive heart failure. Current methods of treating cardiomyopathies are ineffective and target secondary manifestations of heart disease rather than the mutagenic causes. Our aim is to better understand the pathogenesis of cardiomyopathies by studying a particular disease-associated mutation, R369Q, that resides on loop 4 of the β-MHC structure and is implicated in actin-myosin interaction. Here, I studied the molecular mechanistic effect of the R369Q mutation on β-MHC and its role in the DCM phenotype through in silico molecular dynamics experiments comparing the mutant myosin structure to the wild type. Particularly, I examined the flexibility of loop 4, coordination of the nucleotide binding pocket, and electrostatic interactions along the actin interface. We expect to see decreased electrostatic interactions along the actin interface as the loss of a positive charge in this mutant will reduce the affinity of myosin for the negatively charged actin. This work provides a strong foundational hypothesis for the atomic-level impact of the R369Q mutation on myosin dynamics and will be used to design a therapeutic small molecule as well as corroborate in vitro experiments testing the biochemical function of mutant myosin in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Thus, computational and experimental approaches work in synergy in the comprehensive project, informing each other’s results to get a better picture of R369Q’s role in cardiomyopathies at a molecular level and forming a framework for future analyses of β-MHC mutations.

One third of patients in America diagnosed with depression or anxiety are resistant to treatment, creating an urgent need to develop improved therapeutics. In such disorders, motivation to seek rewarding outcomes (‘approach’ behavior) is commonly reduced while apathy (‘avoid’ behavior) is often increased, suggesting that circuitry regulating approach-avoidance (Ap-Av) behaviors may become disrupted in these disorders. Previous research in our lab showed that neurons that release the endogenous neuropeptide nociceptin in the paranigral ventral tegmental area (pnVTA) become highly activated when a large amount of effort is required to obtain a reward, and that activation of these neurons drives avoidance behavior. To investigate pnVTA nociceptin circuitry in directing approach and avoidance behavior simultaneously, I am using NOPLight, a nociceptin biosensor, in an Ap-Av task to record nociceptin release in vivo during Ap-Av decision making in mice. The Ap-Av behavioral assay I am using utilizes a head-fixed apparatus with an aversive LED light positioned at the mouse’s eye level and a sucrose sipper positioned at their mouth. At variable time intervals, either a tone indicating a sucrose reward outcome or a tone indicating an aversive light outcome will play. After the tone plays, the mice will have a short response period where the number of times they lick the sucrose sipper determines the magnitude of the outcome (more licks = more sucrose reward/aversive light, less licks = less sucrose/light). Thus, this task includes simultaneous approach and avoidance components that collectively influence decision-making, and it gives us insight into the role of nociceptin in regulating these behaviors. This research is clinically significant as it can help us understand possible mechanisms underlying the expression of symptoms related to motivation in psychiatric disorders and identify new therapeutic targets to treat them.

Automatically, we consider genes as existing solely to serve the host. Aiding in host survival allows the host to reproduce, causing these genes to propagate in the next generation. However, the existence of selfish genes, which ensure their own survival at the cost of their host, brings this conventional wisdom into question. One such type of gene is called a toxin-antidote (TA) system, which express a molecular toxin and its antidote, the latter of which prevents death, but actively kills hosts that express toxin without co-expressing antidote. The aim of this project is to develop a mechanistic understanding of one invertebrate TA system called PEEL-1/ZEEL-1, which expresses an evolutionarily novel, transmembrane toxin protein PEEL-1, and a transmembrane antidote protein ZEEL-1. Data shows that PEEL-1 co-opts a protein called PMPL-1 to kill animal cells; however, PEEL-1+PMPL-1 do not kill yeast. By identifying why PEEL-1 doesn’t kill yeast, we gain insight into PEEL-1’s toxin mechanism. One hypothesis is that it kills by inducing osmotic stress, which yeast may be invulnerable to due to their cell wall designed to resist osmotic stress. To test this, we’ve expressed PEEL-1, PMPL-1, and PEEL-1+PMPL-1 in separate yeast cultures, chemically degraded their cell walls via Zymolyase treatment, and screened for growth. Preliminary data showing that the digested experimental culture (co-expressing) experiences little post-treatment growth compared to the digested controls (non co-expressing) supports the osmotic stress-related mechanism of PEEL-1 toxicity. Our work in identifying PEEL-1 toxicity will define the first mechanism of an animal TA system and guide our understanding of the different ways that protein-driven cell death can develop in nature.

Substance use disorder (SUD) can be defined as the misuse of pharmaceuticals, illegal drugs, and alcohol, and suggested to be due to the co-opting of existing pathways in the brain controlling natural reward. The endogenous opioid dynorphin (dyn), signaling via the kappa opioid receptor (KOR), has shown great promise in being targeted as an effective treatment strategy for SUDs. However, the role for dyn-KOR signaling in natural reward seeking and specifically, the location of its action in the brain are unknown; information that is critical in informing intervention times for treatment. The Dorsomedial Striatum (DMS) is an area of the brain where dynorphin is ample, and it’s crucial for reward-seeking behavior. However, what dyn is doing to influence these behaviors is unknown. To study natural reward-seeking, we set up an operant behavior task where the mice learn to nosepoke (seeking) into an “active” port for sucrose (reward), whereas nosepoking into an “inactive” port yields nothing. While wild-type mice learn this behavior, we observe that mice lacking DMS dyn are slower and do not seek as many rewards upon learning. This suggests that DMS dyn is necessary for reward-seeking behavior. To determine if the converse is true, we artificially boosted DMS dyn release using in vivo optogenetics. We show that when we stimulated dyn release during reward delivery, mice enhance their seeking behavior. Hence, we hypothesized that dyn may help shape the value of a reward, thereby impacting seeking. We designed a reward devaluation task, where animals are given free access to the reward prior to a session of reward-seeking, to decrease/devalue their seeking. We observe that wild-type animals decrease their responding, but animals lacking DMS dyn don’t devalue their seeking. Altogether, we reveal that dyn transmission in the DMS during reward shapes seeking, thereby enabling value-guided flexibility of reward-seeking behavior.

Mitochondria are the main oxygen consumers in eukaryotic cells and as such are the primary organelles affected by oxygen deprivation, hypoxia. Hypoxia alters the size and shape of mitochondria (so called mitochondrial dynamics) but the responsible mechanisms and their role in hypoxic cell death is unknown. The Crowder lab has recently discovered that a mutation in the Mechanistic Target of Rapamycin Complex One (mTORC1) protein Raptor confers hypoxia resistance. mTORC1 is a master regulator of metabolism and is known to affect certain aspects of mitochondrial biology. In this work I show that disrupting mitochondrial dynamics with mutants in mitochondrial fission produce hypoxia resistance but that mutants with altered fusion have normal hypoxic sensitivity. I have built compound mutants containing both fission and fusion machinery mutants together with the hypoxia resistant Raptor mutant. Using these mutants, I am testing how Raptor controls fission and fusion and whether either is required for its hypoxia resistance. Our preliminary findings indicate that the hypoxia resistance of the Raptor mutant does not require FZO-1-mediated mitochondrial fusion. By exploring the interaction of mitochondrial fusion and fission with Raptor, we are beginning to understand how these important organelle and metabolic regulators combine to control hypoxic cell death.

In the early stages of pathological hypertrophic cardiomyopathy, stress-induced signal transduction promotes the addition of new contractile units to maintain tensional homeostasis through poorly understood mechanisms. Our previous data shows that tension can be changed by altering the contractions within the cells. Inhibition of contraction by expression of D65A cTnC (point mutation on the calcium-binding site of troponin C) results in complete myofibrillar disarray. The mechanism behind the maintenance of myofibril and passive tension seen in these non-contractile CMs is not explained. Data has shown that microtubules provide resistance in the cell and go through rounds of rapid growth or disassembly based on the cell’s need which could be caused by the change in tensional homeostasis. With this in mind, my goal was to determine the role of microtubules in maintaining tensional homeostasis in response to changes in internal tension in CMs. Here, wild-type human induced pluripotent stem cell-derived cardiomyocytes were transduced to express cardiac troponin C with point mutations L48Q (hyper-contractile), I61Q (hypo-contractile), and D65A to study the effect of varying levels of contractility or internal load on microtubules. It was found that microtubular remodeling occurred where the noncontractile CMs had an increase in microtubule density. These noncontractile CMs were then cultures on nanopatterns that provided external tension via topological cues. In addition to the myofibrillar alignment seen previously, it was observed that microtubule density decreased further confirming that microtubules play a compensatory role in these CMs. The next step is to determine if a similar increase in microtubule density observed in the 2D culture is also seen on a tissue level by developing engineered heart tissues. This new data gives insight into how the microtubule remodeling in non-contractile and dysfunctional cardiomyocytes maintains tension in the early stages and its possible role in myofibril formation.

Unprecedented technological growth is upon us. Augmented reality (AR), a technology that enhances our capabilities, is one of the most promising examples. My research shows that AR is often a cost-effective alternative to more widely used technologies such as cell phones and personal computers. It is also seen to be more accessible. Additionally, AR has the potential to be more environmentally friendly, safer, and more efficient than current technology. In this literature review, I attempt to answer the question, "How could AR replace current technology to increase capability and accessibility?" Results and experiments have shown that AR is becoming more capable and cost-effective. This pattern indicates that AR will be adapted more to increase our capabilities as a species. Greater adaptation of AR has correlated with more equity amongst the population, giving equal opportunities for a wider range of abilities and socio-economic status. This is evident with a substantially greater student-to-teacher ratio in schools as well as a much greater potential to train many new individuals in professional careers than would exist otherwise. All of this is done while drastically reducing or even eliminating safety concerns. My research has shown a positive growth and implementation pattern that indicates a future inundated with this technology will soon be on the horizon, if it is not already. This implies a future where AR will help us develop a more equitable society.

Kaposi’s Sarcoma (KS) is among the most common tumors in central Africa and AIDS patients and Kaposi’s Sarcoma-associated herpesvirus (KSHV) is the etiologic agent of KS. While all herpesviruses are capable of both lytic and latent replication programs, KSHV is predominantly in the latent state in the main KS tumor cell,the spindle cell, a cell expressing markers of the endothelium. There is limited viral gene expression during latency so it is difficult to target the virus directly. Therefore, our approach is to target host cellular requirements for KSHV latent infection. Previously, the Lagunoff Lab performed a genome wide CRISPR-Cas9 screen targeting over 18,000 human genes to identify cellular genes essential only to cells latently infected with KSHV. CYP27A1, a gene that encodes a member of the cytochrome p450 family, was one of the top hits identified in the screen. It specifically encodes sterol 27-hydroxylase, which is an enzyme involved in the breakdown of cholesterol. I hypothesize that CYP27A1 is an essential gene for survival of KSHV latently infected cells, as there is evidence that cholesterol is antiviral and KSHV is known to regulate cholesterol. I have successfully cloned CRISPR guide RNAs targeting CYP27A1 into lentiviral vectors, transfected 293T cells with the vectors to make lentivirus, and transduced human tert-immortal endothelial (TIME) cells with the lentivirus to create CYP27A1 knockout cells. ICE analysis was used to confirm strong knockout of the CYP27A1 gene. Currently, we are infecting these cells with KSHV to determine if CYP27A1 is required during KSHV latent infection using cell survival and cell proliferation as readouts. I expect that knockout of CYP27A1 will result in cell death in endothelial cells latently infected with KSHV. By determining genes necessary for KSHV latency, we hope to identify potential therapeutic targets for KS tumors.

The HIV global pandemic has claimed more than 40 million lives so far, and it is an ongoing worldwide health crisis as more than 1.5 million people acquired HIV in 2021 alone. A major barrier towards a cure for HIV is the presence of replication-competent latent HIV-1 proviruses in reservoir cells, which contribute to viral persistence despite years of antiretroviral therapy. One approach to control the latent HIV reservoir is silencing HIV transcription to prevent reactivation. A comprehensive list of host factors that support latency reactivation has not been yet identified. Here, we conduct a CRISPR-Cas9-mediated gene knockout using guide RNAs packaged into budding HIV virions, serving as a readout to identify cellular factors that promote HIV latency reactivation. We used the dependency factor library (HIV-DEP) that contains hundreds of genes involved in proviral transcription. This library was transduced in bulk into two J-lat cell lines, T lymphocytes cell lines which serve as HIV latency models, and we screened for genes that, upon knock-out, prevent latency release after treating the cells with latency reversal agents (LRAs). We identified 47 genes that, when knock-out, prevented HIV latency reactivation in both J-lat models. Our findings suggest that host genes indispensable for HIV transcription also play an important role in HIV latency control. Importantly, these candidate genes could serve as targets for HIV therapeutic intervention.

Research demonstrates that acute and chronic stress can both reduce as well as potentiate an animal's drive to seek reward. In humans, anxiety disorders are also highly comorbid with substance-use disorders. A better understanding of the underlying circuitry connecting these behaviors is necessary for developing better treatment strategies. Neuropeptide S (NPS) acts to reduce anxiety-like behavior and drives drug-seeking through activation of its cognate Gq-coupled protein receptor, NPSR1. The peri locus coeruleus (periLC) produces NPS and sends projections to the orbitofrontal cortex (OFC), a region dense with expression of NPSR1. Although we know the NPS population is involved in cue-reward processing, the elucidation of specific NPS/NPSR1 neuronal circuits correlated to observed behaviors have yet to be documented. In preliminary studies, my mentor, Dr. Kasey Girven, successfully identified a projection from NPS neurons in the periLC to the OFCNPSR1 neuron population. Currently I am investigating the role of the OFCNPSR1 population in drug-seeking behaviors. In my preliminary work I utilized NPSR1-cre mice that expressed a cre-inducibleGCaMP6s in the OFC and a photometry fiber implant above the expressing population to examine the OFCNPSR1 activity during a fixed ratio one task constructed to incentivize an active nose poke with 10 seconds of access to a 10ug/ml fentanyl solution. Using this paradigm combined with fiber photometry, I was able to test the effects of oral fentanyl self-administration on OFCNPSR1 neurons and I uncovered a bidirectional response to delivery of the conditioned stimulus (enhanced activity), and fentanyl reward (quiescence). This experimentation seeks to further our findings on the role of NPS transmission in the OFC and its involvement in drug-seeking behaviors.

Cannabis use has dramatically increased in response to legalization in the U.S., with total sales in the U.S. jumping 46% from 2019 to 2020. ᐃ9-tetrahydrocannabinol (THC) is the primary psychotomimetic compound in Cannabis and has been shown to modify memory and motivation, processes mediated by the prefrontal cortex (PFC) brain region. I sought to test the effects of THC on PFC activity during appetitive Pavlovian conditioning in mice- a behavior in which a subject learns to associate a non-rewarding stimuli to a reward. THC acts on the endocannabinoid (eCB) CB1 receptor (CB1R), a presynaptic signaling protein responsible for modulating neural activity throughout the brain, with robust expression in the PFC. To monitor neural activity during behavioral trials, we implanted optic fibers into the PFC and virally expressed biological sensors: GCaMP6f to track Calcium activity, and the novel GRABeCB2.0 to measure eCB activity. VGAT-Cre and VGLUT1-Cre animals were presented with a house light prior to a sucrose reward to observe the neuronal GABAergic and glutamatergic activity during the conditioning, respectively. After 5 days of conditioning, I administered vehicle or THC (i.p., 5 mg/kg) to observe behavioral and neural effects of THC. We observed neural activity that transferred from the sucrose reward to the house light cue suggesting these neurons encode for this learning. Endocannabinoid activity also transitioned from sucrose reward to the house light cue suggesting cannabinoid involvement in regulating this association. THC pre-treatment reduced licking and motivation for sucrose while modifying neural activity without eliminating it. This provided much needed insight into the formation of memory during learning and reward motivation under the effect of THC.

Δ⁹-tetrahydrocannabinol (THC) is the primary psychoactive compound found in Cannabis sativa and acts on the cannabinoid-1 receptor (CB₁R). Given its well-documented analgesic effects, THC’s therapeutic value in treating pain such as those associated with motor neuron disease states, muscle spasticity-related pain, chronic pain, and muscular sclerosis has gained traction. THC’s psychotomimetic locomotor impairing effects causes patients to cease treatment. However, this relationship between THC and locomotor control is poorly understood. To address this, we are investigating THC’s effects on Pre-Frontal Cortex (PFC) neural activity during natural, unprompted movement behavior in mice. The PFC historically is known for its role in executive function but is also a target for THC’s psychotomimetic effects. We expressed GRAB_eCB2.0, an endocannabinoid biosensor, or GCaMP6f, a Ca²⁺ biosensor, in the PFC and recorded neural activity through fiber photometry during uninhibited movement behavior. We found a novel, THC- and locomotion-dependent transient of Ca²⁺ and endocannabinoid activity in the PFC at the initiation of movement. I investigated the activity of glutamatergic and GABAergic neuron activity in the PFC by utilizing genetic mouse lines and found the Ca²⁺ activity transients were primarily driven by the GABAergic interneurons that constitute 20% of the anatomical population. I hypothesized that this is due to THC-dependent activation of the CB1R on distinct GABAergic interneuron subpopulations in the PFC, which would disinhibit glutamatergic activity and in turn promote spontaneous movement. I utilized in situ hybridization to examine colocalization of CB₁R with distinct GABAergic interneuron subpopulations. We found that while CB₁R does, in fact, colocalize with GABAergic interneurons, there was no differential localization between subpopulations. Overall, this project furthers our understanding of the ways in which THC modulates neuronal activity and locomotive behaviors.

Southeast Asian communities in the United States and in Asia have a preexisting notion that whiteness equates to beauty and that fair complexions are standard. The colorist mindset is implicitly taught through the media, magazines, workplaces, and most commonly the average Asian family household. Teachings of anti-Blackness are introduced from a young age by advertising the use of whitening products, avoidance of the sun, or not being allowed to wear any clothing that exposes skin because of the risk of tanning. This generational upbringing negatively impacts the way young Asian Americans view themselves, view others, and continues the cycle of internalized colorism and anti-Blackness. My research explores how cultural upbringing, generational trauma, and family dynamic influences the understanding of anti-Blackness being something ingrained into Southeast Asian communities. Through conducting interviews and surveys, I examine how colorism is represented and inherently taught within these communities and the ways it has negatively impacted younger generations in terms of self-image. Rather than presenting statistical research, my research presents in the form of a magazine consisting of stories, original art, and portraiture. By incorporating my skills as a photographer and designer, my research encourages audiences to engage on a more interpersonal level to confront the ways colorism exists in our communities and how we can begin a journey of healing and unlearning our prejudiced biases. The results of this project are displayed through a collaborative, ongoing magazine where the interviews are transcribed as articles with photography and original artwork connecting to the theme of colorism and anti-Blackness. This magazine uses the research collected from the interviews and surveys to produce a design piece that shares experiences, allows audiences and participants to be vulnerable, and explores how socio-political topics can be translated through art/design.

This installation is part of ongoing research I am conducting through my interdisciplinary art practice, blossoming out of a fascination with people, their immediate experience, language and healing. When someone interacts with a space or object, whether it is a work of art, another person or the natural world, the felt sense in their body changes, sometimes in subtle and transient ways, other times more obviously pervading. Shortly after absorbing, expressing or repressing the feeling of the moment, language is often added in an attempt to identify the intangible experience we are having. This process happens almost instantaneously. So why does this matter? This matters because artists, similarly to therapists or children, have the unique potential to meet people and moments exactly where they are, an act of empathetic courage that is so desperately needed on a large scale in today's world. Does the potency of a moment change in an environment where words and other technologies are very intentionally omitted or integrated? How do the spaces we inhabit enforce monotony and isolation? Can prolonged attention empower individuals to feel seen and connected to places and people they were otherwise unassociated with? These are some of the questions I am diving into. This project hypothesizes that understanding is not primarily linguistic, instead an energetic and artistic practice that can be catalyzed with loving-awareness and time as a medium. In this research, I am linking together social action, craft, performance art, science and alchemy in hopes of creating validated individuals and therefore more kind and conscious communities.

One-on-one interviews will be conducted with mixed-race individuals from the Seattle area which will give us the personal stories and histories of individuals not typically heard from which we will then transcribe and encode. We will search for patterns on who storytellers tend to be (grandparents, aunts/uncles, etc.) as well patterns about the methods that stories were told through, and how those differing methods impact how participants identify today, as well as how that affects how they interact with the world around them. These will be used to draw conclusions about how having multiple methods of storytelling influences identity formation. We expect to find that non-white parents and grandparents tend to be the people to pass on ways of being, and that mixed race adults believe that although they may have experienced discrimination due to their mixed identity, the insight gained from having a mixed background is overall beneficial. We also expect to find that there may be a tendency to identify more with other mixed people rather than people of any of their discrete ethnicities or races. Thoughtful research and discussion of mixed-race identities can reduce the stigma of being mixed race, making studies like these relevant and necessary. This research will positively contribute to the overall scholarly community by providing information into diverse identify formation, as well as benefit individual participants by providing them a safe space to share their history.
 

An estimated 5-8% of the American population have cerebral aneurysms, showing higher rates of development in patients with common risk factors like hypertension, smoking, and family history of cerebral aneurysms (CA). This study focuses on understanding the causes of aneurysmal subarachnoid hemorrhage (aSAH), where a CA ruptures, resulting in bleeding in the brain. Endovascular coiling is a minimally invasive surgical treatment method for aSAH. Unfortunately, up to 30% of endovascular coiling treatments are unsuccessful, leading to aneurysm recurrence, growth, or rupture. The risk of these outcomes can be predicted using Computational Fluid Dynamics (CFD), a tool that quantifies the hemodynamic environment by solving the equations of motion for a fluid. The CFD simulations calculate factors significant in predicting the effectiveness of coiling treatment including flow rate, wall shear stress, and pulsatility. In this project we have studied the effect of using patient-specific blood viscosity values (the resistance of the blood to fluid flow), that have typically been standardized for all patients in CFD simulations. We have analyzed the effect of using patient-specific blood viscosity on pre-treatment patient-specific computational fluid dynamics simulations of endovascularly-coiled cerebral aneurysms. Preliminary results show that there is an expected improvement in CFD simulation predictive power of treatment effectiveness when patient-specific blood viscosity values are used. We hope to improve the predictive power of CFD simulations regarding the treatment outcome of aneurysm coiling, allowing us to better predict aneurysm recurrence, and eventually guide treatment outcomes.

Pain comes in different types and sizes and affects each of us in different ways. How we perceive pain-related stimuli is highly dependent on our internal brain states. While our pain perception changes in different brain states, little is known about how the brain mediates these changes. Neuromodulators (neurotransmitters, neuropeptides, and related G-protein coupled receptors) are used to elicit a change in neuronal activity, and changes in pain perception often result in behavioral change. Thus, neuromodulation is likely a crucial part in understanding behavioral shifts during brain states shifts. We focused on the locus coeruleus (LC), which uses norepinephrine (NE) as its primary neurotransmitter. NE is highly involved in the modulation of arousal, attention, stress, and pain perception. LC also has broad projections across multiple brain regions, because of this and LC’s use of NE, we hypothesize that LC acts as a hub, modifying behavior depending on the internal brain state. We modeled two representative brain states by exposing animals to either chronic social isolation or exercise via a running wheel. We focused on pain events, as their induced high arousal state is linked with increased NE levels. The pain-related assays include the Von Frey test, which measures mechanical sensitivity, formalin injection, which induces an inflammatory pain response, then the tail flick and hot plate for thermal nociception. LC neuronal activity was measured via cFos expression shown by immunohistochemistry, while NE signaling was measured using fiber photometry with genetically encoded sensors. By using pain-related behavioral assays, LC neuronal activity during pain response, and noradrenergic signaling in our two brain state models, we aim to learn how LC-NE systems affect pain perception during these brain state changes. We believe these methods will help elucidate how change in neuromodulatory signaling levels in the LC mediate behavioral change during brain states shifts.

The Puget Sound region of Washington State has the largest urban ferry system in the world. The most traffic occurs within the central corridor routes, connecting various communities on the Kitsap Peninsula to the metropolitan area of Seattle. These routes are frequented by commuters as they provide a direct connection to the labor market. Traditional land-based modes of commuting, such as by personal vehicle or bus, are shown to negatively impact the commuter. Factors of these traditional commuting environments, such as perceived lack of control and unreliability of the route, act as stressors that can harm the commuter’s well-being in both the short and long term. Many aspects of the ferry environment, however, might avoid these typical stressors. I administered a survey to workers who commute from the Kitsap Peninsula to the Seattle area by a passenger ferry route, and collected data on stress, the perceived reliability of the route, and the positive and negative factors of the commuting experience. Preliminary results suggest that while ferry commuters experience stress due to the unpredictable nature of their route, other factors of the ferry environment cause the ferry commuter to feel overall less stressed than the typical commuter. I aim to supplement previous anecdotal evidence that ferry commutes do not harm worker well-being to the same extent as traditional modes of commuting. Given the unique experience and lessened detriments of ferry commuting, this study can advise researchers to consider ferry commuters as their own distinct class separate from other categories of commuting, and consider how their unique experience can influence transportation policy recommendations and challenge the traditionally negative perception of the modern commute.

Kānaka Maoli (Native Hawaiian) health practices have been significantly impacted by colonialism, beginning with the illegalization of practices like hula, lā’au lapa’au, ‘ÅŒlelo Hawai’i (Hawaiian language), and the growing inaccessibility of cultural foods. Increasing attention has been paid to creating culturally-grounded interventions that address these disparities, which have proven to be effective in increasing health outcomes in Indigenous communities. This study aims to understand the potential benefits of Papa and Pohaku, a culturally-grounded family intervention created and led by esteemed elder Uncle Earl Kawa’a through Keiki O Ka ‘Ä€ina (KKA). KKA is a non-profit organization built to perpetuate Kānaka Maoli culture for ‘ohana (families) and keiki (children). Uncle Earl Kawa'a, a respected kupuna (elder), leads courses on creating a papa (pounding board) and pohaku ku’i ‘ai (stone pounder) to promote healing and wellbeing through grounding participants in cultural practices. To understand the impacts of the intervention, two focus groups were conducted with participants and KKA staff. Questions explored the benefits of participating in Papa and Pohaku, specifically its impact on relationships and traditional Hawaiian knowledge. Our team used Collaborative Qualitative Analysis, which is a structured and rigorous method of conducting inductive thematic analysis. All research team members identified as Indigenous, consisting of one faculty advisor, one doctoral student, and two undergraduate researchers. Respondents reported the intervention positively impacted the following: 1) participants’ pilina (relationships) with their ‘ohana, partners, and with other participants; 2) appreciation for the huaka’i (journey), or process, and commitment to future growth; and 3) understanding of and connection to mo’oemeheu (Kānaka culture). The findings of this study indicate the various benefits of culturally-grounded family-based interventions and a greater need for the availability of culturally-grounded interventions for Indigenous communities.