Inherently, a work titled Pornocrates exudes scandal, intrigue, and allure. Created in 1878 by Felicien Rops, the gouache and pencil piece was displayed at the Les XX exhibition of 1886. Felicien Rops was a wealthy Belgian-born artist who collaborated alongside the likes of Claude Monet, Odilon Redon, and Pierre-Auguste Renoir. Rops’s works articulated more diabolical motifs and grotesque sexual imagery than both his Impressionist and Symbolist contemporaries. As a result, his artworks are far less circulated and research surrounding Rops is scarce. This does not mean he was the only artist creating taboo media, however. Decadent art which depicted debauchery, excess, and sin was fairly present, especially in France. Rops collaborated with many playwrights, authors, and poets at the end of the 19th century, and was surprisingly popular among wealthy socialites of Paris and Brussels. Rops participated in many coalitions of artists across Europe, but the Les XX society is arguably one of the most significant. Artists, authors, and musicians exhibited their works and celebrated individualized aesthetics rather than a unified, nationalistic style. The group rejected a specific political or regional identity, which contributed to the notion of Belgium as a center for the Avant Garde. Rops’s display of Pornocrates elicited shock and commotion among the artists for its uncensored, sexual critique of high society men and women. This project aims to synthesize some of the scholarship surrounding the display of Pornocrates and serves to further highlight the significance of Decadent art in the Fin de Siècle period.

Between 1527 and 1587, three major pattern books were published in Venice: Giovanni Antonio Tagliente’s (c. 1465-1527) embroidery pattern book, Matthio Pagano’s (1515-1588) cutwork lace pattern book, and Federico de Vinciolo’s (active in Paris c. 1587-99) needlework lace pattern book. These three books represent the first of their respective types, and the beginning of a growing social attention toward lacemaking and its use as textile ornament. Each book provides patterns for different types of needlework: embroidery as applied decoration to fabric, cutwork (punto tagliato) as an evolution of embroidery, which involved cutting away the foundation fabric and filling it in with design, and finally, needlework lace (punto in aria), as freely formed design without a fabric foundation. The Venetian attention to dress and outward physical expression necessitated the use of increasing amounts of ornament—prompting the evolution of needlework into punto in aria designs which the city became famous for. Relevant through its additive aesthetic, public demand and commercial opportunities, lace became an essential export for the city of Venice and many women participated in its production, whether within their private homes, in workshops, or in convents. This essay will explore Venetian negotiations with myth and materiality through lacework as an ascendant medium, examining sixteenth-century pattern books and conduct literature as sources of tension and syncretism between written codification and social practice.

Dopamine (DA) neurons found in the ventral tegmental area (VTA) are associated with reward feedback, and dysfunction in DA circuitry is associated with disorders such as Parkinson’s, schizophrenia, bipolar, and addiction to drugs. To adequately treat these diseases, we must have a more complete understanding of how dopamine contributes to emotional processes. This research project addresses this issue by investigating neuropeptide regulation of dopamine neurons in the VTA. The bed nucleus of the stria terminalis (BNST) is a brain region that expresses many neuropeptide genes and sends strong projections to the VTA. We utilized Cre driver lines to isolate neurons that produce the peptides Neurotensin, Neurokinin B, and Corticotropin Releasing Factor. We injected a virus into the BNST that induces the expression of a light activated ion channel and allows us to stimulate axon terminals in the VTA. I then conducted behavioral experiments to assess the effects of activating these peptidergic inputs. Dopamine-dependent behaviors relating to pleasure, reward, and anxiety were measured through the behavioral tests of Real Time Place Preference, operant conditioning, and Open Field respectively. Most likely due to low expression, my behavioral analyses did not yield statistically significant results. Moving forward, it may be necessary to increase viral titer for wider expression. In the future, I intend to use CRISPR/Cas9 technology to isolate neuropeptide function from fast neurotransmitter release in these circuits. This research, by producing findings that help explain how neuropeptides modulate DA neurons, has the potential to generate advances for the understanding and treatment of dopamine-related psychiatric disorders.

Epigenetic proteins modify the chromatin structure to manipulate gene expression, and dysregulated epigenetic modification in cells has been linked to cancer formation. Previous studies on young female Drosophila have shown that after injury, germline stem cells (GSC) are capable of entering and exiting a protective state called quiescence. When exposed to ionizing radiation (IR), the apoptotic differentiating daughter cells send a protective signal to GSC, resulting in GSC quiescence. This survival behavior of GSCs validates them as a potential model for cancer stem cells, which are a subset of tumor cells that are capable of withstanding traditional chemotherapy through reversible quiescence, resulting in future tumor relapse. To identify genes required for GSC survival, we performed a spatially restricted RNA interference (RNAi) screen. Here we show that two members of the repressive epigenetic regulator complex PRC1, Pc and Sce, are required for entry, while demethylase Utx is required for exit of GSC quiescence. Notably, PRC2 dependent H3K27me3 marks are required for PRC1 function, and Utx is required to erase these PRC2 dependent H3K27me3 marks. Importantly, we detected around a 3-fold increase in H3K27me3 marks in GSC following IR, suggesting that the repressive PRC1-PRC2 dependent complex is critical for entry, and elimination of PRC2 dependent marks is critical for exit from the quiescence state. Furthermore, we show that Trx, a writer enzyme which promotes euchromatin formation through H3K4me1 addition, is required for GSC exit from the quiescent. These data suggest that reversible quiescence in GSC is controlled by specific epigenetic states. In the future, more work is needed to investigate gene specificity of the epigenetic regulation.

Induced pluripotent stem cells (iPSC) have a high potential, for they can be differentiated into any cell type for regenerative medicine, drug discovery, developmental biology, and disease modeling. However, iPSC’s and their differentiated progeny display an undesired variability in their shape, contractile properties, growth rates, etc. Identifying subsets of phenotypes in iPSCs and their differentiated progeny will allow us to optimize tissue models for research. Here, we generated a rainbow reporter line in iPSCs that can track individual cells as they clonally expand and differentiate while providing phenotypic information. Knocking in four copies of a cassette containing three distinct fluorescent proteins allowed the expression of up to eighteen different colors. However, not all colors were present in equal proportion, increasing the probability that distinct lineages could have the same color. To achieve an equal color distribution, colored cells were isolated by sparsely plating a culture of mixed colored cells. After a week of expansion, individual colonies were picked and imaged under a spinning disk microscope to determine the color of the colony and whether it was single lineage or mixed. Viable cell lines were isolated and frozen in stock. These cells will be examined for markers of cell proliferation, pluripotency, apoptosis and quantitative RNA expression analysis to confirm that the color barcoded iPSCs act the same as non-engineered iPSCs. To date, we were able to create eight color barcoded iPSC lines for further experimentation, increasing the concentration limit of colored cells in non-colored cells by five-fold. The next step will engineer 3D tissues by growing iPSC-derived cardiac cells in a mold to simulate in vivo tissue development. Colored coded cells will allow us to track how the initial location/physical stresses/phenotype of an iPSC-derived cardiac cell in an engineered tissue determines its tissue layer and cell type.

This research focuses on dissecting the molecular mechanism of cardiac regeneration in the animal model, zebrafish, upon a myocardial infarction like injury. Zebrafish are one of the few vertebrates that can fully regenerate their hearts after an injury in 30 days. This phenomenon is not seen in humans, who generate scar tissue after this injury with reduced circulatory efficiency. However, there is evidence that neonatal mice under 7 days old can regenerate their hearts, but this is lost upon adulthood. Determining this pathway is the first step to develop therapeutics in order to provide relief to people suffering from cardiac injuries. In this research, we used chemically ablated transgenic zebrafish to generate a 30% injury. We determined that upon an injury, both the Wnt pathway and the mTOR pathway are sequentially activated and upregulated to restart cardiac proliferation to regenerate the heart. Wnt pathway proteins like Axin and β-catenin are activated 3 days post injury and mTOR proteins like pS6 are activated gradually over 7 days post injury. The inhibition of the Wnt pathway using DKK showed a downregulation of the mTOR pathway and downregulation of cardiomyocyte proliferation. Inhibition of the mTOR pathway using Rapamycin also stopped cardiomyocyte proliferation from occurring. Mass spectrometry data showed a decrease in glutamine and an increase in leucine during the proliferative phase. Since leucine is one of the activators of the mTOR pathway, we see that the glutamine-leucine transporter is also upregulated post-injury. Thus, we show that heart regeneration in adult zebrafish occurs via cardiomyocyte proliferation by using the Wnt and mTOR pathways to upregulate cardiomyocyte proliferation upon injury.

Helicobacter pylori is a gastric bacterial pathogen that infects more than 50% of the world's population. H. pylori colonizes and persists in the harsh, acidic environment of the human stomach yet it is not an acidophile. The ability of H. pylori to successfully colonize the stomach is dependent on several factors. This includes helical shape that is thought to enhance initial colonization, urease-dependent ammonia production which works to neutralize the pH, and chemotaxis driven motility that enables the bacteria to navigate down the acidic gradient towards the neutral gastric mucosa, its preferred niche. Studies show that flagella-driven motility plays a key role in the pathogenesis of H. pylori; motile strains display higher infection rates and colonize the stomach longer in germ-free piglets compared to non-motile strains. Although the antrum serves as the preferential colonization site due to its lacking acid producing parietal cells, H. pylori can migrate and colonize other sites. In this project, we used a soft-agar assay to characterize the spreading motility of a collection of 42 H. pylori clones isolated from a single host at two different timepoints and from different gastric regions. Our results show that there are quantitative and qualitative differences in the spreading motility of these clinical isolates. The average halo diameters measured are collectively higher in the curved rod isolates from the earlier timepoint compared to helical and rod-shaped isolates from the later timepoint. The clones also display variable chemotactic patterns in soft agar assay with most strains producing concentric rings and a few strains forming uniform halos characteristic of nonchemotactic mutants. By assessing the differences observed, our study explores the factors that impact motility, such as flagellation and cell shape, in order to provide insight into these traits that best equip H. pylori to persist in the stomach. 

The Polycomb Repressive Complex 2 (PRC2) is an important epigenetic remodeler in developmental transitions and cell fate determinations. PRC2 is responsible for the addition of H3K27me3 marks that repress developmental gene expression. The catalytic subunit of PRC2 is the methyltransferase (Enhancer of Zeste 2) EZH2 which binds to EED (Embryonic Ectoderm Development) to methylate H3K27 on gene promoter regions. To investigate the requirement of PRC2 in different developmental transitions, a computationally designed protein was utilized to inhibit EED-EZH2 interaction. The novel designed protein is named EED binder (EB) and competes over endogenous EZH2 on the EED binding cleft with 300 times greater affinity than endogenous EZH2. We cloned EB-GFP under heatshock inducible promoter and injected this construct to one cell zebrafish embryos to generate a germ line transmissible insertion. To study the requirement of PRC2 in early developing embryos (0-3dpf), we applied heatshock (HS) on EB-GFP positive and negative embryos. Western blot analysis revealed global downregulation of EZH2 and H3K27me3 in EB-GFP positive, but not control embryos. Additionally, Co-Immunoprecipitation experiments showed EB-GFP binding to EED. Finally, to test the requirement of PRC2 in caudal fin regeneration, adult (5 month old) EB-GFP positive and negative animals were fin-amputated and the regeneration growth rate was measured for 14 days. Our results show that EB-GFP positive fish were able to regenerate their fins faster, resulting in a large fin size compared to either negative or non-HS clutch mate. Overall, we have developed a computer designed inducible PRC2 inhibitory system to study PRC2 function in Zebrafish, at the whole animal level. In the future, we will utilize EB-GFP to explore PRC2 and other epigenetic modifiers that are required for tissue and organ regeneration before and after injury.

Natural proteins often assemble into various complex geometric structures based on their interactions with each other. The King Lab at the University of Washington's Institute for Protein Design uses the way these proteins behave to develop computational models that enable the design of novel self-assembling protein cages, or nanoparticles. The designed particles are capable of holding and transporting molecules or displaying antigens on their surface, making them effective vaccine candidates. My project involves recovering the solubility of one of these protein cages known as T33_dn2. T33_dn2 is a tetrahedral protein cage comprised of four copies each of two trimeric components known as T33_dn2A and T33_dn2B. While both components can be expressed individually through E.coli before being assembled in vitro, they can also be expressed bicistronically and assemble in vivo. Currently, the use of T33_dn2 as a vaccine scaffold is limited because T33_dn2B is insoluble, and only seems to be stabilized in solution when associating with T33_dn2A. When expressed bicistronically, however, the cage has an extremely low yield. For a protein to be developed into a vaccine, it must be soluble. To recover the solubility and yield of T33_dn2B, I am testing ten plasmid variants of bicistronic T33_dn2. The “original” plasmid consists of one gene coding for a high-expressing cleavable SUMO protein attached to T33_dn2A and another coding for T33_dn2B. The additional nine variants have single point mutations at specific locations on the T33_dn2A gene intended to affect binding strength. After expression, introducing wildtype T33_dn2A in vitro will allow for the formation of T33_dn2. I will be presenting the results of these expression, purification, and assembly tests.

Anxiety symptoms are common in children with attention deficit hyperactivity disorder (ADHD). It is unknown whether the neurobiological origins of comorbid anxiety and ADHD symptoms are shared or distinct. The current study addressed this using an event related electrophysiological potential (ERP) component, the error-related negativity (ERN), which occurs after an individual makes a task error. ERN amplitude has opposite associations with ADHD and anxiety symptoms: it is weaker in association with increased ADHD, but greater in association with increased anxiety. We tested whether 1) anxiety symptoms and ADHD have separate neurobiological origins, indicated by greater anxiety being associated with increased ERN in children with ADHD or 2) anxiety and ADHD symptoms share an origin, as evidenced by no effect of anxiety on ERN in children with ADHD. The current study investigated the association between ERN amplitude and anxiety levels in a sample of 7- to 11-year-olds with ADHD (n = 98) and without (controls; n = 26). Participants completed two ERP tasks of varying difficulty. ERP data were segmented around incorrect task responses, and mean ERN amplitude was extracted. Data on child anxiety and ADHD symptoms was collected via parent report. Linear regression analysis was used to estimate the associations among ERN amplitude, severity of anxiety, and ADHD symptoms. Preliminary results (n = 73) indicated that ADHD symptom severity was associated with smaller ERN amplitude (r =.31, p =.007), but anxiety symptoms were not associated with ERN in the ADHD group. Preliminary results indicated that the ERN is not a marker of anxiety in children with ADHD to the same degree it is in controls. This is consistent with shared neurobiological etiology for ADHD and anxiety symptoms in children, which has clinical implications for conceptualization and treatment of anxiety symptoms in childhood ADHD.

The relationship between humans and robots is full of feedback loops: how our brain processes what we see, feel, and act affects how the intelligent machine reacts and vice versa. Understanding this feedback loop will enable us to design better day-to-day automated systems. For instance, operations performed with surgical robots are guided with the surgeon’s movement. My research in the UW BioRobotics Laboratory investigates how the human brain adapts to technology. I programmed a haptic paddle to apply timed forces on human subjects to investigate how uncertainty, in the form of a disturbance, affects response over time. Understanding how human behavior changes is crucial for improving automated systems that can be simulated by the haptic paddle such as cable-driven surgical robots. We apply the haptic paddle to model forces a surgeon may experience during surgery, such as tough tissues or other disturbances, while operating a robotic surgical device. As we measure the participant’s learning curve over longer periods and more trials, the participant builds a better understanding of how to react relative to their force exerted on to the paddle. By increasing our understanding of how the human brain works, we can begin to improve precision of surgical operation.

After neurologic injuries like stroke, users must relearn how to interact with devices to achieve different tasks. However, hand dominance before and after neurologic injury could affect motor learning and recovery. Understanding a) whether hand dominance plays an effect on motor learning and adaptation of predictive (i.e. feedforward) and reactive (i.e. feedback) controllers and b) how motor learning is impacted by which side was affected by stroke is crucial for personalizing rehabilitation techniques. We are currently investigating the effect of hand dominance on motor learning during continuous tasks for users without motor impairments. 10 participants were asked to play a simple trajectory-tracking game, first with their non-dominant hand and then with their dominant hand. We found that participants’ feedback controllers improved with practice for their non-dominant hand (p=0.005), but their feedforward controllers were unchanged (p=0.88). Furthermore, the participants transferred their feedback controller from their non-dominant hand to their dominant hand (p=0.33). This suggests that the non-dominant hand may be more specialized for error correction and impedance control, and that rehabilitation for users who had a right-hemisphere stroke should focus on improving reactive control.

The mobility of autonomous walking robots is an essential characteristic in their operation. Due to currently imposed constraints in battery technology, the optimization of robotic locomotion for energy efficiency is paramount. Previously, elastic payload suspension has been employed to reduce the cost of transportation in a hexapedal robot. Prior results suggest that the optimal load suspension characteristics are a function of robot morphology and locomotion strategy. A payload suspension system that can be easily adjusted would allow for the accommodation of a variety of these morphologies and locomotion strategies. We have designed a tunable suspension system that will allow for experimentally determining optimal suspension characteristics in a cost-effective manner. The design enables continuous adjustment of the suspension stiffness and damping, so optimal parameters can be determined through hardware experimentation. This hardware experimentation allows for the creation of a numerical model for an oscillating payload’s behavior, which can be compared to simulations. We have completed calculations and design of this hardware, and anticipate seeing that the data collected from its usage will verify the utilization of a haptic testing system in robotics development and allow us to determine methods for calculating the best parameters for elastic payload suspension. This verification of simulated data will allow for further research in developing more efficient methods of payload attachment to legged robots, examination of locomotion when carrying payloads, and design of payload management methods.

X chromosome aneuploidy refers to an atypical number of X chromosomes, differing from two X chromosomes in females, or one X and one Y chromosome in males. Unusual numbers of chromosomes arise from errors in cell division that result in too many or too few chromosomes in a cell, with X aneuploidy reported to occur in around 1 in 1000 births depending on the disorder. X chromosome aneuploidy disorders, such as Klinefelter (47, XXY), Triple X (47, XXX) and Turner (45, XO) syndromes are associated with developmental abnormalities, including cognitive and cardiovascular defects. Our goal is to generate isogenic induced Pluripotency Stem Cells (iPSCs) with different numbers of X chromosomes from patients with X aneuploidy, and subsequently differentiate them into relevant cell types to identify genes affected by aneuploidy. Our lab has previously generated isogenic XXY and XY iPSCs from patients with Klinefelter’s Syndrome by removing the extra inactive X chromosome (Xi). My project is to establish and characterize isogenic lines from mosaic XXY/XY patients, who have a mixture of cell karyotypes. Specifically, I am screening iPSC clones derived from mosaic patients for expression of XIST, a gene expressed only from the Xi, by RT-PCR to determine presence or absence of the Xi. Isogenic control XY iPSC lines derived from the same patient serve as a control to XXY cells, lessening potential for confounding variables due to differences between individuals. Next, I will screen clones with different genotypes for integration of reprogramming vectors by genomic DNA-PCR. Integration-free isogenic XXY and XY clones will be differentiated into cardiomyocytes, neural progenitor cells, and cortical organoids, cell types that are associated with the adverse effects of X aneuploidy. By doing so, we hope to gain insight into gene expression and epigenetic changes associated with X aneuploidy phenotypes in a controlled genetic environment.
 

Fall prevention in hospitals and long-term care facilities in the community setting depends on staff being trained in best practices. However, little is known about the cost, availability, and accuracy of online trainings on fall prevention for nursing staff. The type of education given can be outdated, incorrect or unnecessary depending on what organization that the nursing staff belongs to. The purpose of this research is to identify and evaluate existing online fall prevention training programs and provide the findings to the Washington State Department of Health (DOH) for them to present options to provider organizations as part of the State’s Fall Prevention Strategic Plan. Materials evaluated were previously used for staff education in hospitals and long-term facilities. The criteria for evaluating the websites and educational learning resources (ELRs) were: authority, objectivity, authenticity, reliability, timeliness, relevance and efficiency. Cost was also considered in the evaluation as a factor. Following the evaluation, ELRs were classified by two independent reviewers as either highly recommended, recommended, partially recommended or not recommended for use. From our initial findings, we identified two ELRs that are highly recommended for use, two that were recommended, and three that were partially recommended out of 24 ELRs. After finalizing our results, we will randomly select modules to be evaluated by a group of University of Washington Nursing students for verification of our assessment in reliability and validity of the ratings. We will disseminate findings and recommendations to WA DOH Older Adult Fall Prevention Program and clinical partners through a webinar. This should improve training provided to health providers of care to older adults in hospitals and long-term care facilities. This evaluation method can also be applied to other ELRs to improve the quality of these offerings and assist providers in choosing the best available programs.

In inpatient hospital settings, daily interprofessional rounds are when multiple health care professionals come together as an interprofessional team to create a care plan with patients and their families. Despite the promise of coordinated care during interprofessional rounds, poor care coordination and communication is a major contributor to medical errors, which are the third leading cause of death in the United States. A key literature gap is how to implement sustainable observation and feedback processes for interprofessional rounds. To address this challenge, my research focuses on developing and testing an effective process to evaluate communication during interprofessional rounds. I hypothesized that if pre-health undergraduate students can shadow interprofessional rounds with a structured observation tool, there will be benefits for pre-health students, interprofessional teams, and patients and their families. Specifically, pre-health students participate in an active and engaging shadowing experience while providing interprofessional teams with relevant feedback about interprofessional rounds that can be used to improve interprofessional communication. To test my hypothesis, I used a Plan-Do-Study-Act cycle methodology commonly practiced in quality improvement research. The first cycle tested if a single pre-health undergraduate student could train for and use a structured observation tool, leading to a second cycle which expanded that same process to a cohort of pre-health students. The observation data was analyzed into two categories: 1) quantitative tool topics (with inter-rater reliability data) and 2) qualitative comments. When comparing the quantitative versus qualitative data, the main conclusion was that pre-health students offer insightful and relevant qualitative comment feedback that allow interprofessional teams to reflect on ways to improve their communication during interprofessional rounds. This suggests additional cycles are necessary to test a tool that focuses more on qualitative data. The expansion of my research is key to improve interprofessional rounds in order to continually improve the patient care experience.

Premature infants are commonly born with an inability to feed properly from their mother’s breast, and enteral (tube) feeding is often too stressful on the still-growing body of a premature infant. A recent intervention called Oropharyngeal Colostrum (OPC) administration aims to safely and effectively strengthen the infant’s digestive and immune system, shortening the time to safe enteral feeding. Colostrum, the milk produced right after delivery, is exceptionally high in antibodies and proteins necessary for an infant’s development, and contains an even higher concentration of protective bodies after delivery of a susceptible premature newborn. With OPC administration, colostrum is swabbed on the infant’s cheek or tongue in small amounts on a regular schedule. When the colostrum comes in contact with the tissue in the mouth, the antibodies are taken in and boost the baby’s immune system, heightening resistance to common infections suffered by premature infants, especially in the gastrointestinal region. In 2018, the Lactation Program of the Hospital de la Madre y el Niño (HMN) in Formosa, Argentina began administering OPC to all infants admitted to the Neonatal Intensive Care Unit who were born <35 weeks gestational age and whose mothers consented to the procedure. This research will examine average time to enteral feeding among 400 premature infants administered OPC between 2018-2019. Differences are examined in average time to enteral feeding by gestational age, sex, Apgar score, birth weight, and colostrum administration. The data will be used to determine what characteristics in premature infants cause the most obstacles to enteral feeding. Results will be compared to other studies of time to enteral feeding with and without OPC administration. Results will be shared with the HMN administrators to inform them of the efficacy of the program.

The modern human longitudinal arch is a structural adaptation to bipedal locomotion, rigid enough to be a propulsive lever but flexible enough to stabilize a large body mass over a relatively small base of support. Early hominins appear to have a low or absent longitudinal arch—a condition equivalent to human pes planus. The morphology of the calcaneocuboid joint (CCJ) has been suggested to play a critical role in the stability of the longitudinal arch, but the relationship between articular shape and arch height has yet to be tested empirically. This study examines the covariation of the shape of the CCJ with arch height, via calcaneal inclination angle (CIA). Eleven calcaneal measurements and CIA were measured on weight-bearing radiographs of 103 patients from an urban US trauma center. An equation to predict CIA was determined using stepwise regression analysis using Böehler’s angle, plantar and distal angles of the calcaneal tuber, anterior angle, and the angle of inclination of the posterior talar facet (all p’s<0.01, R2=0.67). Landmark data were obtained from 3D surface scans of the CCJ of 24 calcanei from lower limbs amputated due to infection or acute ischemia. Individuals with bony pathology were excluded from analysis. Data were analyzed in R using Procrustes ANOVA with principal components analysis to explore patterns of variation within the sample. No significant associations were found between CCJ shape and CIA (p=0.51, R2=0.03, F=0.91). Consequently, calcaneocuboid joint shape may not be a useful for interpreting the longitudinal arch morphology of fossil hominin pedal remains.

An estimated 20.6 million people live with Human Immunodeficiency Virus (HIV) in Eastern and Southern Africa. In 2012, the World Health Organization (WHO) announced Option B+ which adopts a single, universal regiment to both treat HIV-infected pregnant women and prevent mother-to-child transmission of HIV. Unlike other HIV treatment options, Option B+ is a lifelong provision of antiretroviral therapy (ART) for all HIV-positive pregnant women regardless of their immune status or viral load. However, I hypothesize that the implementation of prolonged HIV treatment (Option B+) may pose a substantial financial burden for women in very resource poor settings, which could ultimately decrease ART adherence--resulting in no change or even an increase in maternal transmission. This research will examine how economically accessible Option B+ is for expecting mothers in different populations across Eastern and Southern Africa. I am statistically evaluating the cost of ART in relation to household income, living expenses, and cost of childcare at the national level. Future research will use this data to examine maternal transmission risk in relation to economic accessibility, and identify how conceptual frameworks of HIV healthcare are implemented in varied resource settings.

Seasonal and weather change on humans have countless effects on mood, sleep, and diet. Seasonal affective disorder, also known as SAD, is a seasonally linked mood and behavior disorder that typically lasts for four months, starting in the autumn. Similar to depression, SAD involves symptoms of lowered mood, energy loss, fatigue and some atypical symptoms such as hypersomnia, and increased appetite and eating. SAD may have a genetic factor related to serotonin metabolism and melatonin secretion, and rates of autumn-winter SAD may increase with latitude because of the increased seasonal differences in daylight hours. Due to genetic variations among certain ethnic groups, SAD may also disproportionately affect populations who are genetically from southern climates but were raised in northern climates. Yet it is not known how prevalent SAD symptoms were historically in populations adapted to northern climates, and what cultural traditions, if any, these cultures may have evolved in response to SAD. To answer this question, I have conducted a systematic search of ethnographies available through the Human Relations Area Files (HRAF) database. The HRAF databases have indexed and coded ethnographic knowledge across over 400 cultures. I examine the prevalence and traditions of SAD in the indexed ethnographies, and apply these insights to current migrant populations in northern latitudes that may experience SAD.

Recent increases in atmospheric carbon dioxide levels have led to a decrease in the pH of the world’s oceans. These changes in seawater chemistry could have severe consequences for calcifying organisms such as corals. Ocean acidification (OA) poses an imminent threat to the world’s coral reef ecosystems, which could lead to massive reductions in global marine biodiversity. Much of the response of corals to OA is still poorly understood from a detailed physiological perspective, particularly the impact of OA on a fundamental process of skeletal growth called nucleation. Nucleation is the process by which calcium and carbonate ions combine in solution to form a new piece of solid calcium carbonate, which corals use to build their skeleton. While calcification has been measured in corals, the specific step of nucleation has not been quantified and has the potential to be more sensitive to ocean acidification. Coral fragments of Stylophera pistillata and Acropora microphthalma were grown on microscope slides secured in petri dishes in normal ocean conditions suitable for growth. Half of the fragments will be placed into another tank that has constant alkalinity and a set pH of about 7.8 controlled with bubbling air at 800ppm carbon dioxide into the water, the predicted level of atmospheric carbon dioxide in 2100 with the current rate of emissions. This will allow for a big enough change in pH to clearly determine if nucleation is affected by lower pH. With an inverted microscope, the calcifying space was visible through the glass slides, and time-lapses were taken to quantify nucleation rates of corals in the normal ocean conditions. Reefs play a huge role in supporting biodiversity, economies, and population health. Understanding how nucleation rates are changing in variable ocean conditions is a key step in developing new ways to conserve reefs.

Tabletop Role-Playing Games (TTRPGs) are an increasingly popular group activity known both for their collaborative nature and creative demands. Despite the growing popularity, TTRPGs have yet to escape their reputation as a medium for social outcasts to construct escapist power fantasies. The intent of my research is to show that these “power-fantasies,” can actually become a valuable way to explore political and personal identity. I constructed a TTRPG called Blank Spaces, designed for exploring identity, and played with 6 different groups for 22 hours. After each session, I interviewed the players and used recordings of these sessions to qualitatively analyze player experiences. Players were often surprised that they had unwittingly performed meaningful self-criticism and world-criticism, all while simply enjoying a game. This has demonstrated that TTRPGs can be incredibly powerful self-exploration, critisim, and development tools.

Members of the LGBT+ community have long been wanting accurate representation on the big screen. Following the outlawing of “sexual perversion” by the Hollywood Production Code of 1930, homosexuality of any form was only allowed if the character was shown as a villain so as not to encourage their behavior. Traces of this restriction still remain today. Bisexual people in particular are even more problematic with their onscreen images often being either negative or absent completely. This is true even of films created by and for members of the LGBT+ community. To illustrate the pervasiveness of these issues, my research has taken a critical look at four films, Gray Matters, Imagine Me & You, I Can’t Think Straight, and Kyss Mig (Kiss Me), that have bisexual themes and have been celebrated for providing LGBT+ characters with rare happy endings. My project is titled “Happy Endings for Whom?” because I argue that these films perpetuate narratives tying bisexuality with deception and infidelity that are harmful to bisexual people and other members of the LGBT+ community. To do so I break down the plot structures and how the main characters are described and portrayed (ex: Who are the heroes? Villains? How do they identify?). The goal of my research is to expose the power structures that exist within media about and for members of the LGBT+ community and encourage individuals to be more thoughtful audience members and content creators. 

As the 2020 presidential election is fast approaching, per our democratic process, Americans will soon be held responsible to fulfill, arguably, their most important political right. Elections are thought of by many as both a mechanism of accountability and a measurement of performance. Although fair elections are a necessity to uphold democracy, it is essential to recognize that this belief is a misconception. The result of a political election is predominantly decided by the culmination of individual voters making decisions based upon their backgrounds, experiences, and beliefs. To examine this notion further, it is our goal to address the question: How does obtaining a college education affect political voting patterns? To accomplish this task, we have obtained data from the American National Election Survey (ANES). The data includes over 50,000 survey responses, ranging in years from 1948 to 2016, covering topics that include, but are not limited to, wealth, background, beliefs, ethnicity, age, gender, and socio-economic status. With this data, we have performed a regression analysis that is weighted and uses the Taylor Series adjustments to compute design-consistent standard errors. Through our research, it can be seen that the trends reveal a widening gap between educational attainment and party affiliation. Furthermore, those with higher education hold more leftward leaning beliefs, and in turn, are more likely vote Democrat when compared to those with less education. Lastly, the trends over-time reveal a dramatic shift; as, non-college-educated Americans used to primarily be Democrat, while college-educated Americans were mostly Republican. We feel that this no longer holds. This research has implications to demonstrate the importance, or lack of importance, of higher education in terms of the effect and influence it has on real life decisions, which in our case is voting patterns for presidential elections.

Task prioritization, or choosing which order to do tasks, is an essential skill to possess. However, there are no definitive answers to the question of “how” we prioritize tasks. There are numerous factors that are considered in task prioritization, including a vast array of queuing rules (FIFO or First In First Out, EPT or Earliest Processing Time, etc.) However, many of these systematic methods of prioritization don’t account for task complexity. Another common way to prioritize tasks, which will be the focus of this study, is finding the task that seems the closest in time. This perception of Temporal Distance does address task complexity because we balance task complexity and due date when deciding which task is closer or farther away in time. Though individual Temporal Distance Perception is a widely researched topic, there has not been much research conducted on application in a team setting. Prioritizing one’s tasks individually is difficult, but it gets much harder when others’ schedules need to be factored in as well as an individual’s schedule. In addition to this issue of schedule navigation, this research will take place in a multidisciplinary setting, so teammates will not be able to know in detail how long a teammate’s task will take. This too makes it more difficult to choose which task comes first. My research question is this: How does Temporal Distance Perception in a Multidisciplinary Setting affect Task Prioritization in Teams? I am conducting a synthesis of literatures about Task Prioritization, Multidisciplinary Teamwork, and Temporal Distance Perception, as well as creating a data model based off the synthesis. I was sourced the results of team building exercises from NASA’s Human Research Program, which include a variety of real-world examples of how astronaut teams prioritize tasks.

The link between protein sequence and structure is not always apparent. The dogma is that sequence determines structure, but it is not clear how very different sequences can give rise to the same structure. Here, we employ high temperature molecular dynamics unfolding simulations to probe the pathways and specific interactions that direct the folding and unfolding of the SH3 domain, a family of small proteins consisting of two β-sheets arranged to form a barrel. SH3 domain proteins are involved in various functions including protein binding, cell signaling, and nucleic acid modification. The SH3 metafold consists of 753 proteins with the same structure but varied sequence and function. To investigate the relationship between sequence and structure, we selected 17 SH3 proteins with an average pairwise sequence identity of only 27%. Six unfolding simulations were performed for each protein and unfolding transition states were determined, revealing two unfolding/folding pathways. Transition states were also expressed as mathematical graphs of contacts between chemical groups, and three positions in the transition state structure were consistently more connected to the rest of the graph than other nearby positions. These positions represent a folding hub connecting different portions of the structure in the transition state. Analysis of the multiple sequence alignment and covariation also highlighted positions with high conservation due to packing constraints and long-range contacts. This study demonstrates that the SH3 domain can fold through two distinct pathways, but certain folding/unfolding characteristics are conserved independent of sequence and unfolding pathway. By identifying similar interactions, we demonstrate how different sequences can have the same influence on folding pathway and final structure.

 Hoarseness is a common symptom of multiple laryngeal diseases such as inflammation, paralysis, neurologic disease, or laryngeal cancer. Many patients with these diseases are not diagnosed with the correct underlying cause of the hoarseness early enough. Therefore, healthcare providers need better methods to screen for and evaluate different types of hoarseness. Currently, a combination of tools are used to evaluate voice disorders in specialty clinics such as patient history, perceptual voice evaluation, and laryngoscopy. We want to better understand how providers with different medical backgrounds evaluate patients with voice complaints. We are most interested in seeing how history, perceptual voice evaluation, and laryngoscopy impact decision-making and diagnosis. In addition, our group has developed a machine learning algorithm that analyzes voice to detect the presence or absence of a laryngeal mass. We want to see if this algorithm could be clinically useful for generalist providers. To address these questions, a group of clinician evaluators including general practitioners, otolaryngologists, and speech language pathologists, will be recruited remotely. Subjects will be asked to complete an electronic questionnaire with patient case scenarios, asking them to evaluate hoarse voice samples and laryngoscopy exams, with and without case history. For perceptual voice sample evaluations, clinician performance will be compared to the algorithm’s classification of whether a hoarse voice is from someone with a laryngeal mass. From there we will see if clinician detection of laryngeal masses from voice could be improved with this algorithm. If the algorithm has better performance than clinicians, then it may be clinically useful as a screening tool in the future. Our results will help us understand how evaluations for hoarseness are done and can be improved.

Cells use a variety of mechanisms to regulate gene expression. One way cells control gene expression is by forming structures such as DNA loops to position genes in 3D space near regulators. My lab is developing tools to synthetically control 3D genome structure and assess the relationship between positioning and expression. Our strategy is to engineer DNA loops by fusing DNA binding domains to targeting domains that dimerize and bring the two sites together. To ensure the binding domains bind to each other to form a loop, we are developing an allosteric sensor of DNA binding where the dimerization motifs are only active when bound to DNA. Without this switch, the looping interaction would be outcompeted by free binding domains that are not attached to DNA. We used LOCKR, a bioactive protein switch comprised of a protein cage that switches to an ON-state in the presence of a key protein. These proteins are tethered to DNA using dCas9 as a programmable binding domain. I have tested different system parameters, including the length of the linker between dCas9 and the key protein, for increased activation of green fluorescent protein, a reporter gene that is expressed when the switch is activated. My data show that the switch is effective with a variety of linker lengths. I am also exploring other parameters for optimization, such as changing the relative orientation of the dCas9 complexes. Exploring these parameters is important because the switch might be sensitive to small changes in structure or orientation, and we want to identify the optimal arrangement of dCas9 and switch proteins for effective switch function. After this system is optimized, we will be able to direct our efforts toward looping DNA, which will allow us to address broad questions about the relationship between gene position and expression.

One of the Ultracold Atoms Group’s themes is to study the interaction between trapped ultracold atom mixtures. Certain experiments, such as the study of spin-dependent Feshbach resonance, requires us to select one specific nuclear spin state of the atom from the mixture. This process is achieved by the optical Stern-Gerlach technique, where we use the laser to produce the magnetic field gradient. However, this technique requires us to use the imaging path with relatively poor imaging quality due to, for example, vibrations of the optics. When we normalize these atom images, these vibrations introduce misalignment between images and thus unwanted noises. Hence my project is focused on stabilizing the imaging process by a software implementation of imaging alignment scheme. The code I developed can be seen as the analog to the inner product of two vectors, which characterizes the extent of misalignment between two vectors. This scheme has increased the efficiency of the experimental procedure and at least doubled the signal-to-noise ratio. Its easy implementation provides another route to reduce noises in the data of similar experiments. 

In this project, we consider two particle scattering with an arbitrary finite-range potential interaction, contained in a volume by a harmonic trap. The properties of this scattering system are then studied through Effective Field Theory. From this analysis, we attain general relationships among effective range parameters in various dimensions. This project is an extension of confinement induced through periodic boundary conditions studied previously. From this work, calculations can then be made to deduce properties of multi-body condensates. Currently, we already have shown the general relationships between effective range paramaters in toroidal compactified spaces. 

Social determinates of health are such factors as lack of education, low income and low social support that can lead to health disparities. Presence of health disparities can lead to poor communication between patients and providers and consequent poor health decisions among patients. Marginalized populations such as racial/cultural minorities, elderly and the mentally ill are shown to face many of these unique challenges when accessing care. This study aims to determine the ways in which the healthcare system can better support the needs of these safety-net patients. In Washington state, electronic surveys were administered to patients accessing services at a community health center that offers healthcare to disadvantaged and low-resourced communities. The questionnaire was a 47-item instrument that evaluated patients' perception of health services recieved. We were tasked with conducting a qualitative evaluation of a single item on the questionnaire that asked about patients' perceptions of how the healthcare system can best meet their healthcare needs. Using a thematic analyses approach, we evaluated survey responses from 61 patients by reviewing responses for codewords, codeword clusters, and finally, themes that accurately captured perceptions. Two themes were derived in response to the query of how the healthcare system can best support healthcare needs. (1) Accessibilty of affordable resources. (2) Availability of healhcare resources that can support decision making. These finding show that even with access to health resources, low-recourced populations still face barriers to fully gaining access to affordable care to meet healthcare needs.

Recent studies reveal differences in the number of calories consumed by women residing in rural communities compared to individuals living in urban areas. Research suggests that there are many physical activity (PA) barriers for these women. Certain environmental and social factors hinder PA engagement, including environmental safety concerns as well as lack of PA facilities. For the current study, survey administration commenced on September 2018 and concluded October 2018. Researchers partnered with Grays Harbor Community Hospital to administer a 32-item electronic survey to N= 237 female employees. Inclusionary criteria included, English-speaking and self-identify as overweight/obese. My research team conducted a thematic analysis of a single open-ended question, “Are there any barriers to you engaging in any type of physical activity?” and the respondent was asked to explain the barriers as a follow-up. In total, 60 women completed the health surveys, (25% response rate). The mean age of respondents was 53.2 years of age, with a mean BMI of 32.2 (obese status), and majority were healthcare professionals in the hospital. We analyzed the data by conducting iterated evaluations of responses to the question and then deriving themes to characterize respondents’ perspectives. In response to the question of barriers to PA engagement two emergent themes were evident (1) daily life obligations (family and career hinderances) and (2) acute/chronic health concerns that limited physical activity engagement (arthritis, fibromyalgia and back pain concerns). In view of our findings, rural women appear to experience barriers to physical activity engagement. These barriers can help us to identify strategies that specifically target the health needs of residents in rural communities.

Safety-net systems in the United States are crucial in ensuring that low-income and low-resourced, marginalized communities have access to healthcare. Many of the individuals accessing safety-net clinics within their communities are transgender, people of color, and/or Spanish speakers who often-times have unique healthcare needs compared to the general population. The aim of this study is to understand which needs of safety-net patients are not currently being supported by community health clinics, so that they can be considered and implemented in future healthcare frameworks. Our study team reviewed data from a sample of 61 patients from community and human services centers in Washington, DC. The patients who agreed to participate and met the eligibility criteria were taken to a private room where they completed a 47-item self-administered questionnaire. Using thematic qualitative analyses, we evaluated the responses to a single item addressing patients’ perceptions of how the healthcare system can address their healthcare needs. Themes that emerged from our analyses of the single item include the following: (1) safety-net patients seek more resources to support decision-making, (2) patients require accessible and affordable healthcare services, (3) transgender patients require improved gender-related healthcare, and (4) Spanish-speaking patients require more attention and compassion from clinical staff. These findings imply that even with access to healthcare, certain populations continue to face significant barriers to achieving good health outcomes within the safety-net system.

In the United States, women in rural communities face a lot of barriers to healthy eating due to their food environments. Many barriers are associated with busy lifestyles, cost of purchasing healthy foods, lack of healthy food options, and lack of will-power which can lead women to eat less fruits and vegetables and consume more unhealthy foods. The purpose of my project is to evaluate the barriers to unhealthy eating among a sample of rural middle-aged women. Partnering with Grays Harbor Community Hospital, a 32-item electronic questionnaire was administered to N = 237 female employees and 60 surveys were received back (25% response rate). Inclusionary criteria included, English-speaking and self-identifying as overweight/obese. My research team did a thematic analysis of a single open-ended question from the questionnaire, “Are there any barriers to you eating healthy (at least 2 servings of fruits and 3 servings of vegetables each day)?” The respondent was also asked to explain what the barriers were. The mean age of respondents was 53.2 years, they reported a mean BMI of 32.2 (obese status), and majority were healthcare professionals in the hospital. We recorded the responses to the question of barriers to healthy eating and conducted a thematic analysis by deriving codes, codeword clusters and themes. Emergent themes indicated that the barriers to healthy eating among rural obese women were time constraints, affordability, dietary restrictions and nutritional preferences. These findings suggest that perceived barriers are strongly related to their dietary behaviors and this information can offer important insight about helping women to manage their health behaviors.

Ever since they first appeared during the Late Cretaceous, members of the palm family (Arecaceae) have been ubiquitous in the fossil record. Traditionally, palms have been considered a key indicator of warm climates. In addition to leaf macrofossils, fruit, and pollen, palm phytoliths have gained utility as paleoecological indicators. Phytoliths are microscopic silica bodies accumulated in the tissues of many plants. Different plant taxa have unique phytolith morphologies, making them useful diagnostic tools. However, palm phytoliths currently lack diagnostic resolution below the family level, limiting our ability to fully utilize these powerful tools. The goal of our project was to increase this resolution by analyzing the morphology of phytoliths from across the entire Arecaceae family in more detail than has been possible before. We used confocal microscopy to take sharp, high-resolution images of palm phytoliths. Using these images, we took several key measurements, to which we applied multivariate ordination methods. Our analysis allowed us to test how well we can differentiate palm subclades within Arecaceae based on phytolith morphology. Ultimately, we hope to use this information to determine when and where specific clades of palms appeared in the fossil record, increasing our understanding of the evolution of the palm family. This will also allow us to describe past environments in more detail based on palm phytoliths, including estimating more specific climate parameter ranges, and characterizing particular biomes and habitats.

Chondricthyes such as sharks, skates, rays, and chimeras share the slow maturation and low fecundity traits, thus making them susceptible to overexploitation. To ensure the survival of these vulnerable species it is necessary to protect the habitat important to their recruitment. The purpose of the present study was to determine the sediment preference of juvenile Beringraja binoculata to gain evidence as to where their nursery grounds may be located in the Salish Sea. To do this we conducted one-hour filming trials of three four-month old skates in a tank sectioned off into four different sediment size classes. Skate 1 died one-third of the way through the trials so was removed from statistical analysis. However, we found that Skate 2 and 3 preferred to bury and rest in the smallest sediment size of 0.125-0.3 mm grain size with p-values of 0.00126 and 0.0814 respectively. This is consistent with literature on different species of skates around the world. Ongoing research would be valuable to determining the reason behind this preference and to use the information to locate the Salish Sea nurseries.

CD8+ T-cell exhaustion during chronic human immunodeficiency virus (HIV) infection is characterized by increasing expression of exhaustion markers on the cell surface that lead to decreased function, greatly diminishing immune responses and therapeutic vaccination efficacy. Blocking the activation of one such marker (PD-1) using a monoclonal antibody (aPD-1) can help reverse exhaustion. We hypothesized that dosing with aPD-1 would boost the immune system and decrease expression of certain exhaustion markers on CD8+ T-cells throughout infection in order to improve therapeutic vaccine responses. To investigate this hypothesis, we studied how exhaustion changed over time in simian-human immunodeficiency virus (SHIV)-infected rhesus macaques treated with a combinatorial therapeutic regimen consisting of a conserved-elements vaccine to circumvent viral mutants, GS986 to reverse latency, CCR5 gene editing to prevent viral entry, aPD-1 to reverse T-cell exhaustion, and antiretroviral treatment. We characterized exhaustion in peripheral blood mononuclear cells (PBMC) with surface staining and flow cytometry, focusing on markers of exhaustion including PD-1, CTLA-4, LAG-3, TIGIT, and TIM-3 at various timepoints throughout SHIV infection and therapeutic vaccination. We observed higher CD8+ T-cell vaccine responses in animals receiving aPD-1, compared to animals that were vaccinated but not given aPD-1, suggesting aPD-1 dosing improved vaccine responses. We did not observe any significant correlation between PD-1 signaling and expression of other exhaustion markers, suggesting that blocking PD-1 does not reduce exhaustion by decreasing the expression of other exhaustion markers. However, we found a significant negative correlation between CD8+ T-cell vaccine responses and the expression of TIGIT (Spearman r= -0.75, p=0.007). As reduced TIGIT on CD8+ T-cells correlated with increased vaccine responses, there may be a role for dual TIGIT and PD-1 blockade in future studies. Defining the role of CD8+ T-cell exhaustion in therapeutic vaccine immunogenicity and efficacy is crucial to improving combinatorial immunotherapy towards a cure for HIV.

When a child is diagnosed with pediatric cancer, it puts a strain on the whole family, including the marital relationship. However, there is little consensus as to the factors that predict marital adjustment. This proposal examines the relationship between parental emotional styles and marital adjustment in the first year following a pediatric cancer diagnosis. I propose that poorer marital adjustment as well as declines in marital adjustment over the first year of treatment will be predicted by 1) emotion-dismissing philosophies in both men and women and 2) a mismatch in couples' emotion philosophies. These findings would provide insight into the factors that may put the marital dyad at risk, which could inform clinical interventions to better support the families after diagnosis.

I began this project last year as an honors student in the Psychology Department Honors program and under supervision of Dr. Lynn Fainsilber Katz and clinical graduate student, Laina Keim. I independenty discovered a gap in the literature, developed my hypothesis, performed an extensive literature review and wrote a proposal for this this honors thesis. I became a trained and reliable coder for the internationally recognized coding system that is being utilized in this study. This year I have focused on coding our data with our lab team, data analysis with Laina, and now I am writing up results. 

Chronic myeloid leukemia is among myeloproliferative neoplasms that cause an increase of myeloid cells. All CML cases have a reciprocal translocation causing the novel fusion BCR-ABL gene causing the disease. BCR-ABL protein is a drug target and BCR-ABL mRNA is a diagnostic target. The polymerase chain reaction (PCR) is a technique used to diagnose and monitor treatment response in CML. Tyrosine kinase inhibitors (TKIs) have raised the survival rate of CML patients from an average lifespan of 6 years to a normal lifespan. However, monitoring and diagnosing CML patients requires expensive equipment and technical expertise. Medical testing and therapy of CML in lower to middle income countries is difficult, because drugs are expensive and access to sophistical equipment rare. However, when the disease is confirmed by testing, a non-profit organization (the Max Foundation) can provide TKIs free for life through partnerships with pharmaceutical companies. Dried blood spots (DBS) can potential stabilize nucleic acid for long periods of time. This approach has facilitated retrieval of the genetic material on the DBS that can be used to analyze more genetic analysis related to CML response and progression and has reduced transportation costs. In this study, we isolated DNA & RNA from DBS, quantified the genetic material, performed quality control, and carried out DNA & RNA sequencing-based on gene mutation assays. Tapestation and qPCR were performed to check the quality of the genetic material. DNA samples showed a higher DNA integrity number with an average of 7.7. However, the RNA samples demonstrated a lower RNA integrity number with an average of 1.8. The DNA samples had excellent quality whereas the RNA samples had poor quality but were acceptable enough to carry on with our study. These results will help us determine if genetic material on DBS can be used for more genetic analysis.

Currently, there is no clinical best practice in place to support and manage follow up care for previvors, individuals who have tested positive for one or more mutations that increase cancer risk. Patients and providers need support in making complex decisions about follow-up. This project aims to design ‘previvor plans’, modeled after survivorship care plans received by cancer survivors, to help inform and support previvors after they receive a positive test result. Content of the previvor plans includes recommendations for screening, prophylactic surgery, pharmacological interventions, and lifestyle changes based on national guidelines as well as resources to receive further education and support. This project is a part of a larger study which evaluates two different methods of engaging patients in genetic testing for cancer risk across 12 different primary care clinics. Each patient in the study who tests positive for a mutation that increases cancer risk receives a previvor plan tailored with resources and recommendations specific to their test results and location. Use of the plans will be evaluated through surveys and interviews of previvor patients. If successful, the creation of previvor plans will provide a tool which can be used by patients and their providers to manage follow up care after receiving a positive test result.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by the presence of repetitive patterns of behaviors and deficits in social communication and interaction. Attention to social information is a key component of the development of social communication. Previous studies used eye tracking to examine visual scanning patterns associated with social attention in children with ASD and neurotypical children. Eye tracking is used to both identify the atypical patterns of social attention and to predict clinical outcomes in ASD. Although atypical eye gazing patterns are considered as potential biomarkers, researchers commonly consider data loss in eye tracking as error or noise, and rarely investigate it more thoroughly. In this proposal, we hypothesize that loss of data is a potential signature of core social motivation issue when a social video is playing, and, rather than being a nuisance variable, which reflects the broader continuum of social attentional-motivational challenges faced by individuals with ASD. We used eye tracking to confirm previous findings on atypical attention patterns, and further utilize behavior coding to examine the three types of causes of data loss including blinking, non-compliant behaviors, and technical error. We hypothesize that data loss due to blinking is associated with a lack of social motivation and that data loss due to non-compliant behaviors is associated with executive function. Social motivation and executive function were measured by parent reports. Exploring data loss in eye tracking may help reveal comprehensive and fundamental factors of diminished social motivation and neurocognition in ASD.

Hoarseness is a common symptom reported to generalist healthcare providers, with approximately 1% of the clinical population being affected by it each year. It can be caused by multiple etiologies, such as hoarseness due to a cold, acid reflux, or laryngeal cancer. Perceptual evaluation of the voice is inaccurate, and it is therefore difficult to differentiate between hoarseness requiring urgent referral for specialty evaluation (i.e. laryngeal cancer) versus a disorder that could be managed without specialty care (i.e. acute laryngitis). The current gold standard of diagnosis for hoarseness is laryngoscopy, an in-clinic endoscopy recording of the larynx performed by an otolaryngologist specialist. Our research team seeks to improve perceptual voice evaluation by developing and testing machine learning algorithms which analyze voice for underlying pathology, beginning with an algorithm which screens voice for laryngeal masses. We hypothesize that our algorithm will have greater than 80% sensitivity and specificity in the classification of voice samples from patients with laryngeal masses. To test this, we are developing a large, prospective database of voice samples from a laryngology clinic using a smartphone application. Subjects are adult patients presenting to the laryngology clinic, with and without voice disorders, who have had a recent laryngoscopy exam and no laryngeal surgery within the past three months. We are collecting patient history which could influence voice quality, such as age, gender, alcohol use, smoking history, and subject-perceived voice disorder impact. After collection of the voice sample and patient history, cases are classified into underlying pathologic categories. We see recruitment of a well-classified and prospective patient population with a range of voice disorders. This work could lead to improved screening of patients with hoarseness in underserved and primary care settings, and more appropriate and timelier specialist referrals and treatment.

The maternal bond is an intimate attachment between a primary caregiver (PC) and their infant which provides the infant with security, facilitating physical, social, and emotional development. A sensitive and responsive environment, such as the presence of healthy maternal bonds, guides an infant’s neurodevelopment. Changes in mood and emotional state can alter the care a PC provides and cause difficulties in bonding with their infant, impacting the baby’s psychological and physical development. In infants, the mechanisms by which development may be impacted are unknown. Recently, research has shown early atypical attention to visual pop-out in autism spectrum disorders. Attention to visual pop-out describes our cognitive ability to quickly identify differing objects presented among similar looking ones. In this project, we investigated whether PC mental health affects attention to visual pop-out in infants. Participants included 50 infants who were assessed at 6 and 12 months of age. PCs completed the Beck Depression Inventory (BDI) and Beck Anxiety Inventory (BAI) at both timepoints. Infants watched a 4-minute visual pop-out paradigm, which included social (face) and non-social (shape) trials. We assessed responses to visual pop-outs and explored whether BDI and BAI scores correlated with visual pop-out performance at 6 and 12 months. We also investigated whether BDI and BAI scores influenced the development of attention to visual pop-out between 6 and 12 months. We hypothesized infants of primary caregivers who report more (a) depressive and (b) anxious symptoms will demonstrate weaker identification of the pop-out during social trials compared to their peers but be unaffected during nonsocial trials. This study will help deepen our understanding of the impact of maternal depression and anxiety on infant development and help health providers identify and support families.

Recent work published by my colleagues and I showed that diagnostic ultrasound, applied directly over the visual cortex of human participants, increased the likelihood that they would observe visual effects while looking at a visual target, with that likelihood increasing over the course of the experiment. However due to a lack of EEG data, it is impossible to know the biophysical mechanisms and neural pathways that generate the observed effects on brain function. To better understand the full effect and mechanism of our initial findings, I helped develop a surgical protocol for a mouse model allowing for the collection of EEG data while exposing the animal to a combination of light and diagnostic ultrasound stimuli. EEG data was collected from 5 mice using a stimulus paradigm that we believed would increase the rodent's susceptibility to light stimulus. I utilized Matlab for processing, visualization, and statistical analysis of the data to determine the effects and hypothesize potential biophysical mechanisms of the stimuli. My analysis focused on determining whether the ultrasound stimulus successfully increased the susceptibility of the visual cortex, and which brain frequency bands were modulated by the stimulation.

As information processing machines approach the nanoscale level, DNA has emerged as a powerful tool in molecular engineering systems. The specificity and programmability of its hybridization interactions offer flexible and fine-tuned control over reacting species. Among the DNA computing techniques used today, strand displacement circuits are highly popular, with potential applications ranging from disease diagnostics to DNA-based artificial neural networks. The fundamental mechanism of these circuits is the hybridization of a single-stranded DNA input strand to a double-stranded complex which triggers the release of a prehybridized output strand. When released, this output can be detected and used to characterize circuit behavior. The output strands of strand displacement circuits are typically read out using fluorescence spectroscopy. However, due to spectral overlap of traditional reporters (e.g. FAM, TAMRA, Cy5), the number of outputs that can be detected in parallel is severely limited. To address this, we present the use of nanopore sensing technology as an alternative readout device that enables highly scalable, real-time detection and quantification of DNA strand displacement circuits. We demonstrate dynamic sensing of an operating circuit within the flow cell of a commercially-available high-throughput nanopore sensor array (Oxford Nanopore Technologies’ MinION device) and show that strand capture frequency can be correlated to concentration, allowing for direct quantification of desired circuit elements. To investigate this reporter strategy’s multiplexing potential, we present a collection of ten orthogonal circuit output sequences (barcodes) that can be classified at the single-molecule level from raw nanopore signal data using machine learning, with the potential to scale to larger barcode sets. We conclude that nanopore-based detection of strand displacement circuits holds key advantages over fluorescence-based methods for real-time, multiplexed circuit readout on an inexpensive, portable sensor device.

Since the discovery of extrasolar planets (planets orbiting another star) in the early 1990’s, more than 4,000 exoplanets have been confirmed to exist by January 2020 according to the NASA Exoplanet Archive. We are searching for trends between exoplanets’ atmospheric compositions and their physical and orbital properties. To do so, we gather and analyze numerous publications of transmission spectroscopy data on the atmospheres of these planets. The focus parameter space of each search we conduct is expanded incrementally throughout the research process. This expansion requires remaking plots and reanalyzing data, which is a step that has the potential to be simplified. Another problem was needing to input data that would later be used for representation manually. This allowed for the possibility of errors in the data. We also were not able to easily represent all the aspects of the exoplanets we desired to in our graphs and plots, such as stellar type and atmospheric absorption of elements. In order to accomplish these tasks in a more effective and efficient way, the team is automating the data collection, expansion, and representation processes through developing computer programs that are used alongside database queries. This includes developing code that will reduce the amount of human interaction with the data aggregation and representation steps. We will present the improvements introduced with the SQL Server database to store our large data intake and query relationships between planetary properties. Python code is used in SQL Server Management Studio to visually represent these relationships in plots and graphs. This makes for a more efficient pipeline from information intake to representation, which can then be used for planetary analysis. These results will be included in the Habitable Zone Gallery, making it accessible for the community of researchers who wish to use the information as well.

In recent decades, researchers have begun to learn about exoplanets, which are planets that orbit a star outside of our solar system. Exoplanets are very diverse in their properties, for example, their masses, periods, radii, average temperature, and average densities. They also show a variety of elements and molecules in their atmospheres. One of the ways that we use to analyze the composition of their atmospheres is by using transmission spectroscopy. In our research group we are interested in learning whether there are any trends between the physical and orbital properties of the exoplanets, the host stars, and the atmospheric composition of the exoplanets. Our study centers on exoplanets with up to 3.5-days orbits and radii between 1 to 2 times the radius of Jupiter, the so called “hot Jupiters”. The Habitable Zone Gallery is a website which provides information about planetary parameters and how much time each planet spends in its stars habitable zone. The habitable zone is the region in which exoplanets can be found where they have the ability to hold liquid water on their surface. This region is a specific range of circumstellar distance from the host star depending on the host star. We will present the results of this study, utilizing data from the Habitable Zone Gallery, Astrophysics Data System for published data on each exoplanet, and NASA Exoplanet data archive for additional information. We have focused on studying the wavelength range from 3000 to 17000 Å, which is where absorption by sodium, potassium, and water can be found. Any trends between physical, orbital and atmospheric properties will be useful for future selection of targets.

In 2017, Israeli artist Shahak Shapira released his online art installation, “Yolocaust.” In this installation, he downloaded tactless selfies and artsy photographs from people’s public social media accounts which were taken in front of the “Memorial to the Murdered Jews of Europe” in Berlin. He then replaced the background with real images of the Holocaust - inmates on bunks, corpses stacked in piles, etc. His mission was to call attention to the thoughtless and insensitive photographs that people take in front of this key memorial to the Holocaust. In the age of social media, the internet is littered with tweets, Instagram pictures, Facebook statuses, and Tumblr posts about the Holocaust. It begs the question: has social media chipped away at the authenticity at the core of Holocaust remembrance? In my research, I argue that many aspects of social media have been detrimental to Holocaust memory. It has helped to commercialize and trivialize the Holocaust in a new way; however, this phenomenon is not new. For years, the film industry has “dumbed down” the Holocaust in order to make it easy to digest for the average viewer and has greatly romanticized the event in many instances. My project analyzes the Holocaust in film, as well as in Social media in order to argue trends of commercialization and simplification regarding Holocaust memory did not develop newly in the last 15 years. With more digital technology, how we remember this event has been greatly shifted, often at the expense of historical truth, but not as a rule. This project hopes to warn viewers about the price of conveying Holocaust memory through such digital mediums as social media and film, but concedes that the wide-reaching arm of such sources is not always inherently negative and may be exploited as a memory tool in the future.

Despite research supporting the efficacy of certain mental health practices, many mental health care providers in community mental health institutions are not utilizing evidence-based practices (EBPs) consistently as their method towards patients receiving mental health care. This is a problem because the patients in need do not benefit from therapies intended to help them. Clinical supervisors (clinicians who provide clinical direction and guidance to less experienced providers) play an important role in how often and how effective clinicians are at implementing EBP’s in their treatments. The purpose of our project is to examine the feasibility and self-reported usefulness of providing case-based consultation to clinical supervisors in supporting the implementation of EBP. We expect that clinical supervisors will report that they benefited from EBP case-based consultations by having an increase in confidence and competence in supervising mental health care providers in their efforts to use EBP. So far, we have successfully recruited 9 clinical supervisors from 9 community mental health organizations. Clinical supervisors completed a pre-survey that tested their comfortability in supervising mental health care providers implementing EBP. In addition to gathering quantitative data via the survey, we are also analyzing qualitative data from transcripts of the consultation calls with the clinical supervisors. After the 6 consultation calls, we expect that supervisors will rate the calls as high in usefulness. We also plan to learn about any barriers or challenges that the clinical supervisors experienced. With the data that we have collected during this pilot test, we hope to conduct further research testing if EBP implementation to clinical supervisors through case-based consultations creates a downstream effect by resulting in an increased level of competency and comfortability for clinicians.  

There is a long-standing concern that in American society, historical roots, structural racism, and systems of power have perpetuated the spectrum of negative health outcomes in communities of color. As the status quo is maintained, policy and public opinion reflect the continual oppression of minority families. Through my research, I created a visual and multimedia project that will identify the relationship between public policy and the social determinants of health, and how these factors have affected different communities of color within Washington state and the national level. I conducted extensive library research, analyzed and interpreted data, and utilized GIS technologies to visualize the placement of different communities. This work includes identifying historical background information, past U.S. policies, and relevant literature. In this visual and multimedia project, I displayed four puzzle pieces representing different communities (Asian, Native American, Hispanic, and African Americans) on a map, and examined public policies implemented by European colonizers that racialized minorities in unique ways. This chronological project displayed contemporary policies in housing, economic, employment, education, and criminal justice. Overall, I looked at each group's distinct experience of racial health disparities and will use this platform for dialogue to emerge for students and community members on these topics to prioritize the needs, barriers and solutions to confront racism.

Alzheimer’s Disease (AD) is a progressive, debilitating, neurodegenerative disorder where patients lose their ability to think and carry out tasks. This disease is characterized by aggregation of the β-amyloid (Aβ) peptide. Cannabidiol (CBD) and delta-9-tetrahydrocannabinol (THC) are derivatives of marijuana which have been shown to possess neuroprotective properties. Experimental work in this field, is limited in its scope when probing mechanisms driving the phenomenon of Aβ peptide aggregation. Molecular dynamics (MD) simulations have been used to understand the intra-peptide interactions and potential impact of cannabinoids. In order to understand the effects of cosolvent structure on the mechanism of amyloid aggregation, we used classical molecular dynamics simulations of Aβ derived switch-peptides in the presence of model cannabinoids (i.e. CBD and THC). Aβ peptides transform from functional peptides into beta-sheets and therefore impact function within the brain. We tracked beta-sheet formation as a function of time to understand if cannabinoids sterically inhibit interactions between and within peptides. Preliminary results indicate that CBD and THC demonstrate a trapping effect on aggregated peptides. The impact of synthetic cannabinoids are much less understood, prompting additional interest in investigating the interactions among these molecules.

Immunological memory prevents reinfection by a pathogen. This protection is accomplished by memory T cells expressing T cell receptors (TCR) specific for previously encountered pathogen-derived peptides (antigens). Conventionally, memory T cells are thought to be inert during novel infections because there is no interaction between their TCRs with their specific antigens. Despite this, we and others have demonstrated that these T cells (here termed “bystanders”) can be activated by inflammatory signals alone and gain cytotoxic effector function in the absence of TCR-antigen interaction. This study aims to determine how inflammation regulates and attenuates bystander responses and how we can leverage these cells therapeutically. Using in vitro cell stimulations, we found that the inhibitory receptor, programmed cell death protein 1 (PD-1), is strongly upregulated by bystanders after exposure to certain inflammatory cytokines. This finding is unique because the current paradigm is that PD-1 expression is caused by TCR stimulation and PD-1 represents a target to manipulate bystander responses. Further, in mouse models of vaccination, we found that bystander-mediated killing can limit vaccine antigen. We believe that interfering with bystander T cell effector functionality could be targeted to improve antigen-specific vaccine responses. Through understanding the mechanisms that dictate bystander function, we may better modulate bystander T cells function during infection, vaccination, and cancer to improve patient outcomes.

Since they were first discovered in the 1990’s, technological advances have led to a rapid growth in the study of exoplanets, which orbit stars outside of our solar system. Sodium was first detected in the atmosphere of an exoplanet in 2001 with transmission spectroscopy, and a limited but growing number of the 4100+ exoplanets which have been discovered to date have had their atmospheres probed with this technique. Despite the growing number, few studies exist with the aim of comparing the physical properties of different planets and atmospheric information. The goal of our research is to gather previously published data in order to characterize trends relating exoplanet physical properties to their atmospheric compositions. Our research has started with a focus on hot Jupiters: gas giant exoplanets which orbit very close to their host stars. We have expanded the parameter region of interest we are studying, and are working on streamlining the search so that results can be shared more easily with the community through the Habitable Zone Gallery website.

The MAJORANA experiment and its follow-on, the Large Enriched Germanium Experiment for Neutrinoless Double-Beta Decay (LEGEND), search for the creation of matter in the form of neutrinoless double-beta decay, a process that would demonstrate that neutrinos are their own antiparticle and that lepton number conservation may be violated, allowing a deeper understanding of the matter-antimatter imbalance in the universe. MJ60 is a germanium detector used to investigate the low-energy region of the MAJORANA DEMONSTRATOR data, as well as the waveforms produced from incident betas in comparison with gamma events. The latter of these is important for understanding the background of LEGEND: the detectors are submerged in liquid argon, in which beta decays of argon-39 and argon-42 contribute to the background of the extremely sensitive experiment. MJ60 was originally a prototype for the P-type point-contact detectors used in the MAJORANA DEMONSTRATOR. Our group is using this detector to measure mechanisms of energy loss near the detector surfaces by recording events from the nearly monoenergetic beta emissions of metastable krypton-83 (83Kr) at 18 and ~30 keV. Analysis of the waveforms produced from these events, which has been my primary task, will allow us to investigate whether betas incident on our detectors’ passivated surface exhibit markedly different charge collection than gammas, as has been hypothesized. If this expectation is upheld, we will be able to produce a method to identify these events based upon a calculated parameter from the recorded waveforms. Such a parameter will help inform future detector R&D efforts, and will also contribute to background rejection capabilities in MAJORANA and LEGEND.

The COHERENT experiment is endeavoring to detect Coherent Elastic Neutrino-Nucleus Scattering (CEνNS) in several nuclear targets using the Spallation Neutron Source (SNS) at Oak Ridge National Laboratory (ORNL). We search for very low energy (on the order of keV) coherent interactions between neutrinos and atomic nuclei using an array of particle detectors made of different scintillating materials. Neutrinos are fundamental particles under the Standard Model of particle physics that historically have not conformed to theoretical expectations. Understanding their interactions with other particles helps us potentially find other unexpected qualities of neutrinos and in that, discover new physics. At the UW component of the COHERENT team, I along with several other physics undergrads, have just finished characterizing over one-hundred NaI(Tl) crystals to contribute to a large array of detectors at ORNL where they will be illuminated by a strong, pulsed flux of neutrinos born from pions and muons generated in the SNS beam target. For each crystal, I used two characteristic radioactive sources (133Ba & 137Cs) to gather data on each crystal for characterization of their use at ORNL. Each crystal was run for 1.67 hours to find the optimal operating voltage (gain) and to explore the energy linearity of radiation detections at different points in each 7kg crystal (this allowed us to identify if the crystals had any cracks that would interfere with detections). In this talk, I intend to present the statistics on the measured crystal characteristics, including gains, energy resolution, scintillation uniformity as we prepare to ship the crystals to ORNL for installation in the array of other detectors capable of detecting the CEνNS phenomenon.

Mosquitoes primarily navigate using their olfactory system and can use this system to form “memories” that influence their choice in hosts. When a mosquito encounters an odor, information is sent through the antennal lobes in the brain to the mushroom bodies, which are structures responsible for learning and memory consolidation. This odor-learning pathway is mediated by neurotransmitters like dopamine and serotonin. Recent research has shown that the mosquito Culex quinquefasciatus has extremely low levels of dopamine in the antennal lobes compared to other species, and is unable to learn to avoid odors associated with a negative response. This led us to predict that dopamine is essential for aversive learning in mosquitoes. We hypothesized that Cx. quinquefasciatus differed from other mosquitoes in learning ability because they were previously tested in the light and they are the most nocturnal of the originally tested species. To test this hypothesis, we conditioned the mosquitoes in the absence of light in an aversive learning paradigm to measure how frequently they chose to avoid the conditioned odor. An inability to learn regardless of light condition would indicate that the role of dopamine as a neuromodulator in the antennal lobes evolved partly to allow diurnal mosquitoes to avoid defensive hosts. Next, specific neurotransmitters in the antennal lobe were mapped using confocal microscopy, revealing their concentrations which may explain behavioral differences from other mosquitoes. Most mosquito species show some plasticity in host selection, which can lead to the transmission of animal diseases, like West Nile Virus, to humans. Mosquitoes are the world’s deadliest disease vector, killing over 700,000 people globally each year, so understanding how and why this adaptation occurs can help us understand the framework that underlies the spread of mosquito borne diseases and bring us one step closer to solving this global issue.

Since the Industrial Revolution, human-caused (anthropogenic) emissions of greenhouse gases and pollutants, including sulfur, have changed the composition of the Arctic atmosphere. Greenhouse gas emissions and climate feedbacks have resulted in an Arctic amplification, the phenomenon of rapidly warming Arctic temperatures and of sea-ice extent declining at a rate of 7.2% per decade since the 1980s. Other anthropogenic emissions, such as sulfur aerosols, can reflect sunlight and increase cloud cover, temporarily decreasing temperatures. Since the 1980s, clean air policies have reduced the emissions of sulfur aerosols, which have contributed to Arctic amplification. The majority of Arctic sulfate aerosols come from anthropogenic emissions, but natural sources include sea salt, volcanoes, or biological activity. Sea-ice algae produce dimethylsulfide (DMS), which converts to sulfate aerosols through oxidation in the atmosphere. With the decline in sea-ice extent, the habitats of Arctic biota such as algae are diminishing, and it is unclear how declining sea ice will affect biogenic sulfate aerosols and DMS emissions in the future. Here we investigate the relationship between sea-ice extent decline and DMS emissions thorough analysis of an ice core collected in Summit, Greenland to understand the relative contribution of biological activity to Arctic aerosol abundance. To analyze the biogenic sulfate in the ice core, we concentrate ice core meltwater samples, precipitate sulfate in the concentrated sample solution, measure the sulfur isotopes on a stable isotope mass spectrometer, and run GEOS-Chem model simulations to interpret the observed trends. The isotopes of sulfur in sulfate indicate what portion of the atmospheric sulfate aerosols result from biological activity. We hypothesize that biogenic sulfate has decreased with declining sea-ice extent due to the reduction of sea-ice habitats for sea-ice algae. The response of Arctic sulfate aerosol abundance to the decline of sea ice holds implications for the future of Arctic amplification.

Members of the genus Psychrobacter, within the class Gamma-proteobacteria, generally live in very cold marine habitats. These bacteria can be found in Arctic and Antarctic sea ice and sediments, in deep-sea environments, and in permafrost containing relic marine sediments (cryopeg). Each of these environments provides a different combination of temperature and salinity, with different strains of Psychrobacter spp. potentially adapted to grow at different rates depending on environmental source and in situ conditions. I am exploring the growth characteristics of two Psychrobacter spp., each isolated from a different extreme environment. Psychrobacter sp. nov. strain CB7C was isolated from cryopeg brine (originally at –6°C and 140 ppt), and Psychrobacter sp. nov. strain 7E was isolated from winter sea ice brine (originally at –12°C and 128 ppt). We incubated strain CB7C in duplicate at 57 different sets of temperature and salinity conditions, including 19 temperatures, ranging from –7 to 12°C, and salinities of 35, 75, and 120 ppt. The strain was grown in a complex medium, Marine Broth 2216 (at 50% organic strength), adjusted to desired salinity. At regular intervals during the incubations optical density was measured, with cell counts made at start and end. Calculated growth rates and cell yields for CB7C varied across the different temperatures for each salinity. At higher salinity, the temperature at which the bacteria showed maximal growth shifted downwards, a result consistent with in situ conditions (lower temperatures at higher salinities) but novel in microbiology. By conducting similar incubations with strain 7E I will be able to compare growth patterns of the two isolates across a wide set of temperatures and salinities and determine if results with CB7C are singular or represent a more common trait amongst Psychrobacter strains, helping to explain their prevalence under such extreme conditions.

In recent decades, there has been massive growth in consumer demand for products containing live bacterial cultures, or "probiotics", driving a $75B market. Yet, even as market share has expanded, the relative effectiveness of different probiotic products is still not fully understood. Such products require further scientific substantiation before manufacturers can claim health benefits. Few studies have been conducted on how wide-ranging and adverse conditions in the gastro-intestinal tract can influence ingested "pro-biotic" culture function and viability. This research attempts to close this knowledge gap, providing a formal method of characterizing bacterial function under various gut conditions through the identification of biomarkers that are indicative of healthy “probiotic” cultures. L. Bulgaricus, L. Acidophilus and S. thermophilus cultures were evaluated after exposure to conditions simulating major components of the gastro-intestinal tract, their protein expression analyzed and correlated with growth. Simulated colonic conditions maximized bacterial growth, while simulated gastric conditions minimized it. The validity of the experimental model was thus reinforced, as it accurately reflected previous in vivo analysis of bacterial growth in different components of the GI tract. By linking growth and protein expression, the gene, oppa1, was identified as a possible biomarker of cell growth. This gene, activated in conditions that conferred sub-standard growth relative to a positive control, seems to present a key to understanding bacterial population health. This research presents a step forward in the evaluation of the quality of various “probiotic” products by understanding the influence of the human digestive system on live cultures.

Separation of proteins using metal ligand complexes is a well-established practice in the field of bioengineering and biochemistry.  electrospray ionization mass spectrometry (ESI-MS) can be used to identify bio and organic molecules. Previous studies have detected chelated metal ions using ESI-MS, this project focuses on the identification of a metal ligand complex comprised of a tridentate chelating agent Iminodiacetic acid (IDA) and a bidentate ligand, Histidine. By coordinating copper with IDA a binary complex is formed, this allows for the detection of copper by proxy of the IDA. By selecting a unique fragment related to IDA we can target in tandem mass spectrometry (MS/MS) for greater sensitivity; the complex can be selected for analysis out of solution. We will use this novel approach to build a parent-ion scanning technique to monitor metal-ligand complexes extracted from environmental matrices. first the ideal solution parameters are determined to maximize the complex formation and detection of the Cu-IDA complex. So far, a high ratio of copper to IDA coupled with a basic buffer have yielded the best data. Creating a ternary complex comprised of copper IDA and an imidazole ring containing compound, histidine. Selectively tuning to the peaks associated with the copper IDA complex, the ternary complexes can be selected for in depth analysis of its structure and bonding properties. Future work could focus on identification of metal ligand complexes from soil samples with other compounds containing imidazole rings such as the neonicotinoid imidacloprid which has been indicated in bee colony collapse. By chelating solid with IDA complex formed from pentacoordinate copper ions could be detected despite the low relative concentration.

Back contact solar cells improve on the standard geometry by arranging both the positive and negative electrodes on the back of the device. This significantly improves efficiency by eliminating three critical challenges faced by typical solar cells: (1) shading of the active layer by top contacts, (2) the conflicting requirement for conductive yet transparent top contact materials, and (3) difficulty in printing metallic contacts onto the sensitive photoactive layer. Electrodeposition is a disruptive method that offers an alternative pathway for solution processing manufacturing, but is relatively unexplored as a method of fabricating perovskite back contact solar cells. This study focuses on achieving selective electrodeposition of high quality nanoscale electron and hole transport layers (tin and nickel oxides respectively) onto interdigitated silver collectors. In this investigation, a silver working electrode and a nickel or tin counter electrode against a silver/silver chloride reference electrode comprise a three-probe system in nickel nitrate and tin chloride electrolyte baths. The conductivity and carrier mobility of the electrodeposited transport layers are discussed as a function of salt and dopant molecule concentrations in the electrolyte bath. High carrier mobility layers are desirable for increased solar cell efficiency. The microstructure and thickness of the transport layer are discussed as a function of bath temperature, stirring speed, and the magnitude and duration of the supplied current density. The transport layers are characterized using optical profilometry and scanning electron microscopy. Diode devices are fabricated to characterize the electrical properties of the oxide layers. Based on these results, an optimal electrodeposition procedure is recommended for producing high quality nickel- and tin-based charge transport layers for application in back contact perovskite solar cells. Achieving this device geometry via electrodeposition enables the production of more efficient solar modules using high throughput manufacturing methods - bringing novel photovoltaic materials one step closer to widespread use.

Humanity is on the verge of a biotechnological epoch. What this will entail is a union of opposites: biological humans merging with artificial machines. Unity of opposites, or non-duality, is a timeless theme, one that is not only found in the ancient teachings of Heraclitus or in Greek mythology, but also in that of the I-Ching, and the Tao Te Ching. The yin-yang symbol is one of the earliest visual depictions of this. Support for a unified theory of the universe is now being widely embraced; hence the shift from the old Standard Model of particle physics to the new Core Theory. Modern interpretation of what Laozi referred to as, Tao, suggests a process that is characteristic of a double torus, with a cuboctahedron at its heart. Such a synergic principle would not only be observable in the nature of matter, but also in that of the mind. It is of no coincidence that humanity is beginning to integrate with arguably its finest of achievements—tools. Synergic inquiry is the method by which this literature review will be conducted, so as to build the argument that all things are interdependently connected, distinguishable sub-systems of a larger system. This implies that the convergence of human and machine is simply the beginning of a brand new, distinguishable part in that whole. The study of synergetics, in its wider applications, proves to be an invaluable tool to understand the macrocosm and microcosm relationship, and it will no doubt contribute greatly to the symbiotic relationship between nanotechnology and molecular biology, as humanity seeks to build a better world.

Early initiation of antiretroviral therapy (ART) among people living with HIV significantly improves health outcomes and survival rates. Based on these findings, the World Health Organization recommended provision of ART to all people living with HIV regardless of disease status. In 2019, the Government of Tanzania initiated task-sharing to nurse-initiated HIV care as a way to mitigate health systems challenges associated with increasing numbers of patients eligible for ART. As health providers take on new responsibilities, it is critical to assess the quality of HIV care. We are presenting observational data collected during a technical assistance program across 5 regions in Tanzania. Supportive supervision teams visited 20 health care facilities to observe provision of HIV care. During baseline observations, teams used a 41-point checklist to assess individual health care providers administering HIV care. Domains within the checklist included visit type, introduction, adherence counseling, consultation, and communication and support. Following the observations, the teams reviewed individual results with the providers and offered feedback. A repeat evaluation was performed after 3-months. Data has been summarized as counts, proportions, means, and standard deviations. Chi-squared tests of proportions have been used to compare pre/post data. Data analysis has yet to be completed. Findings from our analysis will measure differences in quality of HIV care over time in the context of HIV task-sharing and will provide specific information about domains of quality of HIV care. Results will be shared with the Government of Tanzania to inform future program directions. Quality assessment is important in providing HIV treatment in accordance with national guidelines. Our data analysis will inform future clinical training and health systems monitoring to ensure efficient and effective HIV care in Tanzania.

In order for cells to generate copies of themselves they must undergo a highly complex process called mitosis. During mitosis, many enzymes called protein kinases work together to ensure both daughter cells inherit the correct number of chromosomes when the cell divides. Polo-like kinase 1 (Plk1) is a protein kinase that regulates several events during mitosis including centrosome maturation, spindle assembly, sister chromatid cohesion, and cytokinesis. Recently, the A-kinase anchoring protein Gravin (AKAP12) has been implicated in regulating Plk1 function at mitotic centrosomes. Specifically, loss of Gravin has been linked to defective protein signaling at centrosomes, chromosome misalignment, and increased incidence of micronuclei (small nuclei, an aberration often seen in cancer). However, while previous studies used shRNA-mediated knockdown to reduce Gravin levels in cells, it remains unclear how complete loss of this scaffold in human cells influences mitotic signaling events. To test this, our lab generated Gravin knockout U2OS (osteosarcoma) cells using CRISPR/Cas9 genome editing. First, I employed a combination of immunohistochemical staining and quantitative imaging tools to assess how Gravin loss affected chromosome alignment, micronuclei formation, and gamma tubulin accumulation at centrosomes. I found that loss of Gravin in U2OS and HeLa cells caused misaligned chromosomes and micronuclei. Additional experiments I conducted revealed that Gravin-depleted U2OS, HeLa, and MEF cells presented aberrant gamma tubulin accumulation at mitotic spindle poles. Next, a local drug-targeting approach was used to specifically inhibit Plk1 activity at mitotic spindle poles in U2OS cells. I determined that localized inhibition of Plk1 produced similar mitotic defects as observed in cells lacking Gravin. Collectively, these findings suggest that Gravin is required for coordinating proper Plk1 signaling at centrosomes during mitosis while the loss of this scaffold protein leads to mitotic defects. Future work will uncover downstream substrates of Gravin-anchored Plk1 that becomes dysregulated in cells lacking Gravin.

Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen associated with worsening disease outcomes in cystic fibrosis (CF) patients. P. aeruginosa uses quorum sensing (QS), a cell-cell signaling system, to control expression of a variety of genes including virulence factors. In P. aeruginosa, QS is mediated in part by acyl-homoserine lactone (AHL) signals that can diffuse in and out of cells. Once AHLs accumulate, they bind to a receptor regulator that activates gene transcription. P. aeruginosa has two complete AHL QS systems, LasI-LasR and RhlI-RhlR. The two systems are arranged in a hierarchy, with the las system controlling the rhl system. QS activation in P. aeruginosa is restrained by cellular proteins that dampen the QS response. These proteins, known as “anti-activators”, attenuate QS by preventing receptor activation. Three anti-activator proteins, QscR, QslA, and QteE, have been identified in P. aeruginosa. These anti-activator proteins have additive, overlapping roles in repressing expression of QS gene products in laboratory strains but their role in the QS dynamics of CF isolates is still unclear. This project used standard molecular cloning techniques to delete or overexpress anti-activator genes in a selection of clinical isolates from CF patients. A reporter plasmid with a fluorescent marker was used to track the activity of LasR and RhlR. These experiments were used to quantify differences in QS-controlled gene activation. To test the hypothesis that anti-activators decrease the amount of LasR in the cell, Western blots were used to assess the cellular levels of QS receptors. In strains with deleted anti-activator genes, LasR levels were higher and induction was earlier. Additional tests for phenotypes controlled by QS, such as protease and pyocyanin production, were also performed. Future research should focus on evaluating these effects in additional CF isolates.

Throughout history, human-induced habitat loss, pollution, and hunting have pressured mammals to adapt to lifestyles that limit human interaction. When humans threaten wildlife, a fitness advantage is provided to animals who avoid human interaction. But what about in protected regions where these threats are limited? Cocha Cashu Biological Station, located in Manu National Park, is an example of one of these regions. While native communities continue to hunt within the research station, overall levels of hunting, deforestation, and pollution, are significantly lower than in the surrounding unprotected areas. Our research team chose to design a study in Cocha Cashu to analyze terrestrial mammal distribution in relation to human habitation in areas where human threats have historically been limited. Our hypothesis was that mammal abundance would increase with distance from human habitation. Our study design involved a northern and eastern transect with a near (N), medium-distanced (M), and far (F) trap location. Each trap location had both a sand and camera trap, and data was collected from the traps morning and night for four consecutive days. On the eastern transect, there was a positive linear relationship between distance from human habitation and number of mammals observed (N: 2 mammals, M: 4 mammals, F: 8 mammals). On the northern route, the highest number of mammals was observed at the medium-distanced location (N: 1 mammal, M: 7 mammals, F: 3 mammals). Overall, our data did not support our hypothesis that mammal abundance increases with distance from human habitation in protected areas. Our results do, however, provide a platform for further research on resource accessibility and its potentially larger influence on mammal distribution patterns than the influence of human habitation.

The Cretaceous-Paleogene boundary is one of the most well-studied mass extinction events in history. However, there are still many questions left about what was occurring in terrestrial ecosystems in the 10-15 million years prior to the event. One major question is if theropod dinosaurs were declining in diversity and ecological stability prior to the event, or if they all died off suddenly. This stability is partially interpreted in this project through a geometric morphometric approach comparing Judithian and Lancian theropod teeth. Geometric morphometrics quantitatively analyzes tooth shape based on points called landmarks, that are universal across theropod teeth so variance can be measured and compared. Teeth are digitally imaged and processed with the landmarks to compute and then interpret their shapes in specialized programs. The landmarks measured are based on established standard literature of mesial and distal points of the tooth, the apex, and the mesial and distal terminal denticle. If the finalized analysis of the remaining confirms theropod dinosaurs are shown to have consistent morphological disparity through this time, this will be an indication that this group of organisms were not declining and were settled in their niches prior to the K-Pg boundary. Given that theropods were a diverse group that occupied many niches, as well as being the top carnivores that would have curbed other taxonomic populations, studying their economic stability through the window of deep time lasting from the Judithian to the Lancian immediately before the K-Pg boundary is leading to an invaluable understanding of how this environment was changing during this time.

Today there are more than 5,000 species of extant mammals that are categorized into three major clades: placentals, marsupials, and monotremes. The deep evolutionary history of marsupials is poorly known due to a fragmentary fossil record. In particular, cranial fossils of the ancient relatives of marsupials (stem marsupials) are extremely rare. The few cranial elements of these taxa that have been found are fragmentary, crushed, or missing major elements. Recently, some more complete cranial fossils of stem marsupials have been discovered at the Late Cretaceous (75 million years ago) Egg Mountain fossil locality in northwestern Montana. My research will describe the morphology of a partial skull of a stem marsupial from Egg Mountain. This delicate skull is encased in a hard siltstone, making it difficult to mechanically remove without damage to the fossil. Instead, we used micro-computed tomography (µCT) to scan the specimen block. Then I used Avizo software to virtually remove the rock and expose the details of the encased fossil. From the resulting files, I will study and describe the cranial morphology in detail. Thus far, we have identified the stem marsupial as Alphadon halleyi. With further study, I hope to (1) expand upon current knowledge regarding the morphology of Alphadon halleyi, (2) make comparisons with other stem marsupials and extant marsupials, and (3) more broadly, incorporate our findings into a phylogenic analysis of early metatherians (the clade that includes stem marsupials and marsupials) that will further elucidate the evolutionary history of marsupials.

Previous studies on plant phenology have found that shifts in the timing of life cycle events are connected to changes in the surrounding climate. Alpine and subalpine wildflowers are of particular interest, considering that the initiation of flowering often depends on the timing of snowmelt, which has been occurring earlier as temperatures warm. However, we often lack the information needed to predict exactly how much wildflower phenology will shift in response to warming. Herbaria, collections of plant specimens collected over the last 100-200 years, have recently been paired with climate data (from the geographic locations and dates of specimens) to examine the relationship between climate and plant phenology. I aim to answer 1) How does climate influence the timing of phenological stages in wildflowers? and 2) Do species vary in their responses according to their average bloom time after snowmelt (e.g. early season vs. late season bloomers)? This study involves five alpine/subalpine species that very in the timing of blooming relative to snowmelt date: Western pasqueflower (Anemone occidentalis), Sitka valerian (Valeriana sitchensis), Sickletop lousewort (Pedicularis racemosa), Rainier pleated gentian (Gentiana calycosa), and Glacier lily (Erythronium grandiflorum). I will access specimen data from the University of Washington Herbarium and the Consortium of Pacific Northwest Herbaria. Phenological stage will be recorded with the date of collection, geographic locations will be determined using GEOLocate coordinate and elevation data, and climate conditions will be spatially modeled using ClimateWNA historical records. I predict that there will be an overall trend of earlier flowering for all four species over time, but that species with later bloom times will experience less deviation from their historical average than those with earlier bloom times. This study is significant in providing information that will help the preservation of wildflower meadows in the high mountains of the Northwest.

Triple-negative breast cancer (TNBC), the most aggressive form of breast cancer that accounts for 15% of all breast cancers, tests negative for the three common breast cancer oncogene proteins– estrogen receptor, progesterone receptor and HER2. Without specific biomarkers to be targeted for treatment, TNBC treatments are limited to combinations of immunotherapy, chemotherapy, lumpectomy and radiation therapy. Alternative biomarkers such as the PD-L1, PD-L2 and CD-86 are utilized to explore more specific treatment options with minimal side effects. Detecting biomarkers in the tumor tissue typically utilizes immunohistochemistry (IHC), which exhibits limited quantitative precision, sensitivity and multiplex capability. To address the challenge, multiple reaction monitoring mass spectrometry (MRM-MS) has been developed for quantitating multiple tumor biomarkers simultaneously. Accurate detection of targeted biomarkers on the MRM-MS requires samples with high purity. However, the current microbead immunoprecipitation process results in a noticeable level of background impurity. To compensate for this impurity, the assay requires higher peptide yield by using at least 5 mg of tissue (around 100 µg of protein), which may not be available or can only be obtained with more invasive procedures. Additionally, the current biomarker extraction protocol is a 16 manual-step workflow that takes 38 hours. To speed up the biomarker purification process while using less tissue sample, our group has developed a biopsy tissue processing microfluidic device. Instead of magnetic microbeads, the device utilizes smart polymer reagents with higher capture efficiency to improve the peptide purification process and reduce the required sample size down to less than 5 mg of tissue. The device contains pre-loaded reagents, which streamlines the processes of deparaffinization, cell digestion, protein digestion and peptide purification to only 8 hours with 4 manual steps. The new device with the smart polymer reagents can potentially enable comprehensive tumor microenvironment characterization using MRM-MS with needle biopsy tissue.

Until recently, B lymphocytes have not been thought to inhabit non-mucosal human skin. They have traditionally been viewed as major elements of the immune system strictly confined to blood and bone marrow. This study reports the novel finding of B cell infiltrates in herpes simplex virus 2 (HSV-2) skin lesion biopsies. Motivated by an interest in the underlying immune mechanisms responsible for the wide spectrum of disease seen in those suffering with reoccurring HSV-2 lesions, I examined the immune microenvironment of tissue biopsies from study participants with active herpes lesions and a history of frequent outbreaks. In order to study the local immune response of an HSV-2 reactivation episode, I analyzed skin sections of herpes lesions using fluorescence in situ hybridization (FISH) to demonstrate the cell marker and immunoglobulin expression profiles of skin-resident immune cells present at the time of herpes outbreaks. Through the use of FISH and advanced deconvolution microscopy, I proved that B cells of the human immune system can home directly to upper layers of the skin to fight HSV-2 reactivation. My work reveals the presence of immune cell populations in HSV-2 lesions showing RNA expression for CD20, CD138, CD4, IgG, IgM, and IgA. This study demonstrates that antibody-producing cells are recruited to sites of HSV-2 reactivation, secrete a variety of antibodies at the site of infection, and appear to interact closely with CD4+ T cells directly in skin. B cells may play a crucial role in the immune response required to effectively control HSV-2 reactivation, which has been underappreciated until now. My work suggests that individuals with immune systems that are less efficient at recruiting B cells to the upper layers of the skin to control HSV-2 infection may explain why some individuals suffer more severe manifestations of the disease than others throughout their lives.

Antiretroviral therapy (ART) cannot eliminate latently infected human immunodeficiency virus (HIV) reservoirs, the barrier to HIV cure. A “shock and kill” strategy has been proposed to cure HIV by using latency-reversing agents (LRAs) to reactive latent proviruses and allow for reservoir elimination. Due to the low potency and high toxicity of LRAs, none have yet been effective in reducing reservoir size in vivo. Here, we hypothesize that delivery of LRAs using nanocarriers (NCs) will improve drug solubility and safety, provide sustained drug release, and simultaneously deliver multiple drugs to reservoir tissues and cells. We developed hybrid nanocarriers to incorporate physicochemically diverse LRAs and target reservoirs in lymphatic CD4+ T cells. LRAs were formulated by physical encapsulation or covalent conjugation to the biodegradable polymer (PLGA) core. Drug combinations were evaluated in vitro using a J-Lat reporter cells and validated in CD4+ T cells from virologically suppressed patients. CD4+ T cell targeting specificity was tested ex vivo in non-human primate (NHP) peripheral blood mononuclear cells (PBMCs). Targeting and toxicity were also evaluated in vivo in mice following size optimization for increased passive drainage to lymph nodes. Optimized nanocarriers were used for identification of an LRA combination displaying synergistic latency reversal and low toxicity in vitro in model and patient cells. Long-term and specific activation of CD4+ T cells in NHP PMBCs ex vivo and in mouse lymph nodes in vivo was observed, with significant reduction in toxicity compared to free LRA delivery. This nanocarrier platform targets CD4+ T cells, successfully inducing latency reactivation in HIV reservoirs. The platform additionally enables new solutions for HIV cure with the potential to deliver anti-HIV agents, vaccines, immunomodulating agents, and gene-modifying oligonucleotide drugs.

Scaffold proteins, which assemble enzymes and their substrates into multiprotein complexes, are critical for many cellular functions. By bringing enzymes and their substrates into close proximity, scaffold proteins are thought to enhance the rates of enzymatic reactions. For instance, Axin is a scaffold protein in the Wnt pathway that binds the transcription factor, ß-catenin and the kinase, GSK3ß. This tethering method is predicted to enhance the rate of ß-catenin phosphorylation. An outstanding question is whether a scaffold protein that binds to both GSK3ß and ß-catenin is sufficient to observe these effects or whether Axin has other structural properties that are not currently understood. Although there is no detailed structural information about Axin, it is predicted to be disordered. To address this gap, we have engineered a synthetic scaffold protein that can bind both GSK3ß and ß-catenin and contains a linker that is also presumed to be disordered. Previous work in the lab has reconstituted the reaction between Axin, ß-catenin and GSK3ß in vitro and found that Axin enhances the rate of ß-catenin phosphorylation. Using enzyme kinetics, I have compared the rate of ß-catenin phosphorylation of the engineered scaffold to that of Axin. Identifying the quantitative kinetic comparisons between the engineered scaffold protein with the natural scaffold protein could tell us more about what aspects of a tethering system make it effective at enhancing the rates of reactions. We are expecting to see similar tethering effects between the two scaffold proteins, as their linkers are both disordered and equivalent in length. 

Probiotics are living organisms that when ingested have been linked with health benefits to the gut and improvement of conditions such as irritable bowel syndrome (IBS). The gut contains a plethora of microorganism populations that make up the microbiota. To understand how these populations communicate with each other, and the tissues surrounding them, it is imperative to identify and characterize the method of communication. Extracellular vesicles are one such possible method. Extracellular vesicles (EVs) are lipid-bilayer delineated sacks of material secreted from cells. It has been established that EVs are used as a waste disposal system. However, new research revealed that EVs can be used by the cell for methods of communication. Furthermore, EVs are now being linked to cell-to-cell and cell-to-organism communication. If EVs have been linked to communication, then characterizing them is one step closer to understanding how probiotic bacteria function. Previous studies have mainly characterized EVs by their size and divided them into 3 main groups: exosomes (40-150 nm), microvesicles (100-1000 nm), and apoptic bodies (>2000 nm). However, an analysis of proteins found in these EVs has not been performed yet. Here we compare EV proteins to proteins in the cell, to determine which protein fractions are secreted by cells through vesiculation for signaling purposes. To separate the cellular fraction from the EVs fraction, cell suspensions were centrifuged. First, the cells were pelleted and collected at 300x g. The leftover supernatant was spun at 16,000x g to pellet the EVs. Then, proteins from cells and EVs were solubilized and digested with trypsin. The tryptic peptides will be analyzed using liquid chromatography-mass spectrometry. A comparison of proteins in cells and EVs, and their relative concentrations can help us learn more about how probiotic EVs function in the gut.

Probiotics are found in numerous fermented foods such as yogurt, sauerkraut, and kombucha. Research shows that eating live probiotics are beneficial for the gut because it helps supplement the plethora of native bacteria. Now, these microorganisms are gaining popularity throughout the world—people are ingesting them in the form of food, drink, and even pills. Most probiotics are anaerobic. In fact, excess oxygen can damage organelles, create ionic imbalance and eventually even kill the cells. However, in the process of manufacturing these fermented foods, the probiotics in them often get exposed to oxygen, for example, due to leaks in packaging. Even the FDA does not have a rule about standard manufacturing processes regarding anaerobic conditions and yogurt. This could mean that the probiotics people eat are highly compromised: damaged or dead. I hypothesize that probiotics exposed to aerobic environments for extended periods of time will express more DNA repair enzymes such as DNA pol. 1 and 2, p53 or photolyase. This is because oxygen will cause probiotic cells to change function, and therefore, they will start to express enzymes, such as these DNA repair enzymes, to protect them from oxidative damage. In order to test this, I left probiotics out to oxidize for various amounts of time—0 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, and 1 hour. After oxidizing, I cultured the cells for 16 hours. I then pelleted the cells by centrifugation at 300 x g and washed and lysed them. Then, I tryptically digested cell proteins and analyzed them by LCMS (Liquid Chromatography Mass Spectrometry) for identification. In the future, these proteins can provide insights to how these oxidized probiotics impact the gut. Are they really the miracle microorganisms that benefit the gut, or could their compromised condition end up being harmful?

In trying to understand biology’s dynamic heterogeneity, scientists have sought to recapitulate the spatial complexity and temporal presentation in which proteins are naturally presented to cells. Currently, the most promising strategies in this regard exploit sequential ligation/cleavage reactions, each controlled in time and space using light so as to reversibly immobilize proteins within synthetic biomaterials. Though our lab has utilized these approaches to spatially control complex biological fates with micron-scale resolutions, previous methods suffer from complex syntheses, as well as requirements for specialized equipment and skillsets rarely available in bio-based laboratories. Improving upon these fundamental limitations, our group has developed a scalable system wherein proteins can be bound/released from hydrogels using light, without the need for such expertise/equipment, by being fully genetically encodable. In this approach, biology performs all the modifications necessary to photopattern protein binding to gels, as well as install the reactive species requisite for the protein’s photo-mediated release. We have accomplished this using a photoactivatable protein-peptide ligation reaction developed by our lab, wherein UV irradiation “activates” the protein to ligate specifically with the peptide tag. Additionally, we exploit co-translational chemoenzymatic modification strategies to install a functional handle for tethering the protein into polymeric hydrogels during protein expression. To the peptide tag, we append a photocleavable protein that cleaves when irradiated by visible light, fused to a protein of interest (POI) to be tethered to the hydrogel. Expressing these proteins in E. coli yields the first-ever fully genetically encodable system which can reversibly pattern proteins into hydrogels, by first shining UV light to tether POIs into biomaterials, then subsequently shining visible light to photocleave the protein and trigger POI release. Highlighting the system’s versatility, we demonstrate that the approach is compatible with fluorescent proteins and bioactive growth factors to direct 4D cell fate.

Water-swollen polymeric networks (i.e., hydrogels) provide a structural platform for the manipulation of chemical and mechanical signals that mimics the complex heterogeneous environment experienced by cells in vivo. Photoresponsive chemistries have been of particular interest to this end, as they allow for precise spatiotemporal control of physiochemical properties and, thus, cell behavior. Here, we present a novel protein-based network that will allow for the photo-mediated stiffening of genetically-encoded hydrogels. In this system, we exploit a biochemical technology recently pioneered by our lab in which two pairs of proteins undergo irreversible, covalent heterodimerization after photoactivation. Through the incorporation of an inert, unstructured polypeptide backbone, we have exploited the aforementioned reaction to induce gelation in response to light through the formation of four-arm protein crosslinks. Unlike previous synthetic polymer-based hydrogel systems, this system is entirely genetically encoded, which provides significant advantages in terms of cost, time, and production simplicity. As we intend to demonstrate through photorheometry, this reaction proceeds in a dose-dependent manner, providing step-wise control of both where and when gel stiffening occurs. Such 4D control of a gel’s mechanical properties can be used to influence cell migration, growth, and differentiation, and, thus, could have applications in tissue engineering. Furthermore, we anticipate our system could be utilized to model the stiffening of the extracellular matrix, which is commonly associated with pathologies such as cancer, fibrosis, and cardiovascular disease.

Latinx individuals feel that mainstream mental health services cannot adequately address their specific needs and that non-Latinx therapists may be insensitive to their psychosocial needs. The Latinx population is a rapidly growing population. In 33 states they account for 58% of the population; in the remaining they account for 7%. As a result of increase in population in the United States, there is an urgent need to understand the Latinx culture and create interventions that are effective for Latinx individuals. Studies reveal barriers around therapists lacking empathy towards Latinx clients, and lack of language metaphors in order to translate mental health diagnosis. To address these gaps, the study aims to discover what are the barriers and needed supports to promote culturally competent care by mental health practitioners serving Latinx communities. This study uses primary data obtained through a quantitative survey from mental health practitioners who are serving or have served Latinx individuals. Chi-square and independent samples t-tests were run to examine bivariate relationships between whether trainings related to cultural competence, humility, and diversity were provided and participant perceptions of cultural competence and understanding of the Latinx culture. A preliminary analysis of 18 participants found no significant differences at the p < .05 level. However, trends suggest practitioners who are provided with training across categories of competence, humility and diversity were likely to report feeling culturally competent. Additionally, no meaningful differences were seen in average ratings of knowledge of the Latinx culture. Preliminary findings suggest trainings are beneficial for practitioners working in mental health within the Latinx community, particularly for facilitating cultural competence but not necessarily for increasing knowledge around the Latinx culture.

Many universities do not have separate language courses for heritage language (HL) learners and second language (L2) learners, and over the last decade, an increasing amount of HL students have been enrolling in the same language classes as their L2 peers. How does the same heritage language learner engage in Spanish classroom pair work activities when working with a heritage versus a second language learner? Student engagement drives learning. If one population shows low engagement, it is detrimental to their learning. In this project, we are analyzing engagement as a multidimensional construct which includes the branches of cognitive, behavioral, emotional, and social engagement. The participants in our case studies are HL and L2 learners taking equivalent upper level university Spanish courses, the former in a course designed for HL students, and the latter in one for L2 students. I hypothesize that a heritage language learner will be most engaged when working with another heritage language learner. Their skill levels will be the most similar, so they will have a level of ease and comfort with each other, and will relate to each other more. I am tracking behavioral engagement by number of words, number of turns, overlapping speech, and latched speech, in which there is no pause between the speakers’ turns. Emotional engagement appears as turns including laughter. Lastly, I am marking social engagement through content-related and language-related questions, co-construction of ideas, and repetition of ideas, which may include an error correction. My research will contribute to understanding of interactions between matched and mixed dyads, which is important information for language teachers and the teaching practice.

Kaposi's Sarcoma associated Herpesvirus (KSHV) is an oncogenic human herpesvirus and the etiological agent of Kaposi's Sarcoma (KS), a cancer that afflicts HIV-positive individuals worldwide and is endemic in sub-Saharan Africa. KS tumor cells, known as spindle cells, originate from latent KSHV infection of endothelial cells. We have previously used a phosphor-proteomics approach to identify changes in the phosphorylation state of proteins during KSHV infection. We identified Platelet/Endothelial Cell Adhesion Molecule-1 (PECAM-1) to be significantly phosphorylated during KSHV infection as compared to mock infected cells. PECAM-1 phosphorylation leads to a cascade of signaling that promotes cell adhesion, migration, and cell survival, however, the role PECAM-1 plays to aid KSHV infection is currently unknown. To determine whether PECAM-1 expression and phosphorylation is important for KS pathogenesis, I propose to assess differences in gene expression levels with RT-qPCR, validate phosphorylation levels in vitro via western blot, and generate CRISPR-lentiviral constructs to knock out PECAM-1 and express PECAM-1 isoform variants in endothelial cells. These experiments will paint a more complete picture of how PECAM-1 interacts with endothelial cellular processes during KSHV infection. Many rounds of RT-qPCR have been conducted to asses gene expression levels with highly variable results. This indicates that some uncontrolled factor in the cell culture process is affected PECAM-1 expression levels. I hypothesize that cell density is the cause behind this variability and plan to use western blotting to elucidate differences in PECAM-1 protein levels in low and highly confluent cells.

Honeybees, Apis mellifera, demonstrate an ability to perform visual learning to such an extraordinary extent that it is not typically associated with smaller brains. Those learning abilities may be linked to their degree of sociality as they aggregate into colonies and synergistically work together on tasks. The Social Brain Hypothesis suggests that an organism's level of intelligence is correlated to how social their environment is. Some bees like the leaf cutter bee, Megachile rotundata, are solitary bees who do not live in a hive but instead reside in individual nests. The purpose of this research is to explore the visual learning capability of leaf cutter bees and compare them to the learning capability of the well documented honeybee. The methods used for this study include placing a tethered bee on a free rotating ball that is placed in front of a screen. Two colors are projected onto the screen and the bee controls the movement of the shapes. A positive and negative reinforcement are assigned to each shape. By analyzing how many times the bee picked each color, how fast it responded to the shapes, and how far from a distance it walked to fixate on that color, we can make an informed statement about the solitary bee’s ability to visually learn. We anticipate one of three outcomes, the leaf cutter bee will either have a greater learning rate, less learning rate, or equal learning rate to that of the honeybee. These results will help supplement more data for the Social Brain Hypothesis and allow a deeper understanding of how sociality is related to cognition. This study also has major relevance to pollination practices as leaf cutter bees are used in agriculture, therefore understanding the neural basis of cognition can open the door for new reforms in crop pollination.

Parkinson’s disease (PD) is a common neurodegenerative disorder that is caused by the death of dopamine-secreting neurons in the midbrain. The onset of symptoms such as progressively worsening tremors, movement difficulty, and dementia are thought to be caused by protein aggregates called Lewy bodies, mitochondrial defects, and neuroinflammation. Mutations in the GBA gene, one of the genes responsible for PD symptoms, accounts for 5-10% of all PD cases. It codes for the enzyme glucocerebrosidase which breaks down the lipid glucosylceramide. Expression of GBA in muscle cells reduces protein aggregation in the head and GBA is found in extracellular vesicles (EV). Similarly, expression in the midgut partially reduces protein aggregation in the head. Our goal is to determine if GBA expressed in the midgut can travel in EVs. We have developed a GBA mutant fly model that features symptoms similar to human PD – neurodegeneration, shortened lifespan, motor deficits, and increased protein aggregation. Flies were bred and crossed to the correct genotype, collected and frozen, and processed for analysis. To ensure that GBA is not expressed elsewhere to prevent confounding, we use an Npc1b promoter with the GAL4/UAS system, since the Npc1b gene is only expressed in the midgut. Protein is quantified following gel electrophoresis and western blots. We expect to find GBA in EVs which would help confirm that GBA traveling in EVs is responsible for the reduction in protein aggregation. This is potentially an important feature for the development of treatments for PD, as understanding what causes protein aggregation is a key step in eliminating or reducing it to prevent PD.

The Davis lab is focused on understanding the pathogenic cellular mechanisms causing Parkinson’s disease and other aging-associated neurodegenerative diseases.We are currently researching how GBA affects extracellular vesicle aggregate protein propagation that increases the advancement of Parkinson’s disease. My own research in the lab is focused around the effects of GBA mutations on ceramide metabolism, a lipid in extracellular membranes, and how ceramide levels affect the propagation of Parkinson’s disease. I will use a GBAdel Drosophila model with Carmofur to increase Ceramide levels and determine if any visible changes in extracellular vesicles can be seen. I will also use a neuronal model created from Parkinson’s disease patients to look at and characterize vesicles when ceramide levels are changed. Through these two models I will be able to analyze and determine if variation in ceramide metabolism levels are due to GBAdel and how ceramide levels affect extracellular vesicles related to Parkinson’s disease. GBA is known to encode for GCase, an enzyme that hydrolyzes glucosylceramide into its parts, glucose and ceramide, since GBA directly allows for the creation of ceramide, it is reasonable to believe a mutation in GBA would affect ceramide levels in a Parkinson’s patient. It is also reasonable to assume that changes in ceramide levels will correlate with the aiding of Parkinson propagation, due to ceramide’s role in extracellular vesicle creationWe are using a fruit fly model based on a genetic risk factor for Parkinson’s disease, and a human neuronal model derived from Parkinson’s disease patients with genetic risk factors for this disease. We are currently investigating how this risk factor may influence faster propagation of pathogenic protein aggregation throughout the brain. We hope to identify new possible therapeutic targets to slow or stop neurodegeneration in Parkinson’s disease, where currently only symptomatic treatment exists.

Experimental evolution can determine genetic interactions during natural selection in complex systems. Using whole-genome sequencing of 95 parallel populations of haploid Saccharomyces cerevisiae experimentally evolved for 250 generations, we discovered a possible epistatic interaction between two sets of beneficial mutations. The first mutation is a transposable element (TE) insertion into the promoter of FLO1, giving rise to a cellular aggregation phenotype known as flocculation. The second set are putative loss of function mutations in genes encoding members of the SAGA-complex, which is thought to increase expression of genes proximal to TEs. We hypothesize that without the members of the SAGA-complex, the FLO1 gene will be unexpressed, abrogating the flocculation phenotype. We isolated three flocculant clones with TE insertions from different experimental populations and crossed them with three clones with the deletion in the SAGA-complex. Through meiosis, the yeast sporulated into four cells. The ratios of flocculant to wildtype haploid cells are used to determine an epistatic interaction. A 2:2 ratio suggests a non-epistatic interaction while a 1:3 flocculant to wildtype ratio suggests an epistatic interaction. The project is in the early stages but segregation ratios suggest the members of the SAGA-complex with deletion mutations do not hinder the expression of the FLO1 gene. Our alternate hypothesis is members of the SAGA-complex have no effect on the activation of TE insertions that promote the expression of the FLO1 gene. While the initial hypothesis might not hold, this experiment will give us a further understanding of genetic interactions during evolution.

Biomechanical characterization of human tissue is of increasing importance as new technologies gain importance in modern medicine. Without haptical sensation, measurement of forces associated with surgical techniques will be the primary source of feedback in technology assisted procedures (eg. robotic surgery). An important biomechanical parameter is the suture pullout force (SPOF), the maximum safe force that could be applied to a suture before tearing the tissue.The aim of this study is to analyze suture pullout forces of the pancreatic duct. This data could be used for a wide range of applications and will be useful in understanding how biomechanical properties of the biliary tract change with age, sex, and BMI. Donated organs used in this study were tested within 72 hours of death. Cross sections of the pancreas were prepared and 4-0 sutures were looped through one side of the pancreatic duct wall. The pancreas was held in place while the suture was pulled in tension. The tensile force was measured continuously and the peak force before specimen failure was taken as the SPOF. 103 suture pullout tests were performed on pancreatic ducts from 14 donors. The overall SPOF is 2.87 N ± 1.36 N. The mean SPOF for females is 2.12 N and 3.03 N for males (p=0.019). The SPOF of pancreatic ducts from donors with BMI less than 26 averaged 3.22 N while SPOF for donors with BMI greater than 26 averaged 2.51 N (p=0.045). Donors older than 35 averaged 2.69 N while donors younger than 35 averaged 2.87 N (p=0.123). Preliminary results suggest male pancreatic ducts have a higher SPOF than female pancreatic ducts. As BMI increases, the pancreatic duct SPOF decreases. The effect of age is inconclusive at this time.

Sleep deprivation (SD) is a major health concern in developed countries, especially in the elderly, and is associated with physiological disturbances including cognitive impairment, increased risk of dementia, and leads to the acceleration of neurodegenerative disorders such as Alzheimer’s disease. A logical question is whether cognitive impairment associated with SD can be prevented. To address this question, 22-month-old C57BL/6 mice were sleep deprived during the middle of their sleep cycle for four hours each day for two days. Three days before the SD procedure, treatment was started with the anti-aging peptide glycyl-L-histidyl-L-lysine (GHK) 15 mg/kg per day for five days through the two-day SD procedure. GHK has wound healing and anti-inflammatory properties and readily passes through the blood-brain barrier. Immediately following the last day of SD and GHK treatment, mice were tested for learning impairment using a spatial learning activity which requires them to find the escape hole. Tissues were then collected for neuropathology assessment. Sleep deprived mice treated with GHK consistently found the escape hole more quickly than sleep deprived mice treated with saline suggesting that GHK can prevent the learning impairment associated with short term sleep deprivation. This preliminary observation provides the rationale to investigate the neuro-molecular and neuropathological pathways targeted by GHK including inflammation, oxidative stress and vascular dysfunction.

Astrocytes are characteristic star-shaped glial cells, and they are the largest and most numerous nonneuronal cells in the brain. They are involved in cell signaling, providing nutrients to neurons, removing toxins, repair and anti-inflammation response in the brain. However, relatively little attention has been paid to this cell type compared to neurons in neurodegenerative conditions, such as age-related cognitive impairment and Alzheimer’s disease. The basic question is: what functional changes occur in astrocytes with increasing age that could relate to vulnerability to neurodegeneration. C57BL/6 mice in age cohorts of 8 months, 16 months, 24 months, and 32 months were tested in a radial water tread maze, a well-standardized measure of contextual learning and memory in the mice. Brain tissues were then collected and parasagitally sectioned and stained with GFAP (Glial Fibrillary Acidic Protein), an immuno-reactive marker for astrocytes. In general, 8-month old mice showed very little cognitive impariment while several 16-month old mice showed mild cognitive impairment, but 24 and 32-month old mice showed moderate to severe cognitive impairment. Concurrent with this increase in cognitive impairment was an increase in ImageJ pixel intensity of GFAP immunohistochemistry staining of astrocytes with increasing age. This preliminary observation suggests that astrocytes are predominantly present with increasing age and decreasing cognitive ability and provides the rationale for further investigations into what role these nonneuronal cells are playing in brain health.

Senescence is a gradual deterioration of functional characteristics. At the cellular level, it is defined as a loss of the ability to divide and carryout normal cellular activities that contribute to the health and well being of a cell. In fact, senescent cells secrete inflammatory cytokines that contribute to aging. Therefore, preventing senescence could have an impact on preventing aging. To address this issue, 21-month-old C57BL/6 mice were fed standard rodent chow containing three well-validated anti-aging drugs, rapamycin, acarbose, and phenylbutyrate for 3 months. Each drug targets different but overlapping aging processes. At the end of 3 months mice were tested for cognition and physiological performance and tissues collected for assessing senescence by reverse transcriptase polymerase chain reaction using well-accepted senescence markers including p16. Preliminary observations suggest that the drug cocktail in just 3 months delayed aging by improving cognitive and physiological performance tasks. Preliminary data from liver samples tested by reverse transcriptase polymerase chain reaction showed that mice treated with the drug cocktail had less P16 expression than mice fed a diet without the drug cocktail. More work is needed to sort out how each drug is contributing to this anti-senescence effect, but the preliminary observation suggests preventing or delaying senescence may be an informative target for investigations into delaying or preventing aging.

Lateral roots are branches originating from the main branch of a primary root. They provide stability to the plant, and assist in acquisition of nutrients and water, similar to the primary root. It is well understood from studies in Arabidopsis thaliana that lateral root development is regulated by the plant hormone auxin. A few undifferentiated cells respond to a  pulsatile auxin signal to become ‘specified’ lateral root stem cells, retaining potential to proliferate and ability to differentiate into a lateral root. These specified lateral root stem cells are arrested in G2 phase of cell cycle, respond to auxin signaling and undergo rounds of cell division marking the onset of lateral root development. The focus of my study is to better understand how cell cycle control influences lateral root developmental transitions from undifferentiated to specification to initiation. In particular, I am interested in addressing the question of whether cell cycle arrest in G2 phase crucial for lateral root development, and if so at which step of cellular transitions get altered specification or initiation. An important tool necessary to address this question is the ability to control the length of G2 cell cycle arrest for lateral root stem cells, and we have recently generated transgenic lines for this purpose. I am inducing lateral roots in a number of transgenic plant lines and using fluorescence microscopy to observe the early stage of lateral root development. These experiments allow viewing of both the staging of development and cell cycle stage associated with development over time. The understanding gained from these experiments will help build a framework for how cell cycle contributes to lateral root development, allowing for future genetic modifications to improve root structure in crop plants.

Plants respond to environmental changes by changing their growth patterns. For plants to continue to grow throughout their lives, they are constantly undergoing cell fate determination—how the plant determines the specific fate of new cells they are generating. One example of this is the development of roots that emerge from the primary root, called lateral roots. The number and spacing of lateral roots determine the overall root structure, which determines how well the plant takes advantage of resources in its environment. Auxin is a plant hormone involved in many aspects of growth and development, including the regulation of lateral root production. Using data from an experiment where mRNAs were sequenced from single cells isolated from roots, the Nemhauser Lab identified specific genes that may be active during lateral root development. My research question is: are these genes targets of auxin signaling? To test this, I will use quantitative RT-PCR to measure expression of the candidate genes in wild type and in plants that are deficient in the auxin response pathway. Genes that are targets of auxin should show lower expression levels in the mutants relative to wild type. The more we understand about how cell fate determination occurs in lateral roots, the more we can understand the underlying mechanisms by which plants arrive at their final root structure. Our understanding can then guide engineering or breeding projects, allowing optimal root growth that could drastically increase plant survivability and yield.

Cancers are broadly characterized by changes in cell metabolism. Tumor cells typically exhibit functional respiration and inhibition of electron transport chain can impair cancer cell proliferation. However, certain neuroendocrine cancers can arise from loss of function (LOF) mutations in succinate dehydrogenase (SDHx/complex II), which plays a key role in the TCA cycle and in mitochondrial respiration. SDH, which catalyzes the conversion of succinate to fumarate, comprises four subunits: A, B, C and D. LOF mutations in subunits B, C, and D can promote tumorigenesis and mutations in subunit B (SDHB) are particularly associated with malignant and metastatic neoplasms. Interestingly, SDHB impaired cells show an accompanied loss of activity in complex I, implying that unlike the majority of cancer cells, respiration is not essential and may even be antagonistic for SDHB mutant cancer cell proliferation. Indeed, preliminary experiments indicate that inhibition of complex I can restore proliferation to cells treated with an SDH/complex II inhibitor. However, the molecular mechanisms behind this phenomenon are not well understood. We aim to investigate the metabolic mechanisms by which dysfunctional respiration is essential for the proliferation of SDH impaired cells. We hypothesize that inhibition of respiration in these cells can prevent oxidation of NADH to NAD+ at complex I and alter the redox homeostasis in the mitochondria to support proliferation. Specifically, we will test to see if increasing the NADH/NAD+ ratio is the required function of complex I inhibition that rescues cell proliferation in SDH impaired cells. In addition, we will characterize the metabolic consequences of specific alterations SDH, complex I, and mitochondrial redox state. Results from this study should allow us to delineate the importance of metabolic alterations in SDH mutant cancer cells and potentially help identify metabolic vulnerabilities for treatment of SDH impaired cancers.

Human papillomaviruses (HPV) contribute to approximately 4.5% of cancer cases worldwide. There are currently three vaccines which protect by producing neutralizing antibodies to the structural capsid protein, L1, for each of the most oncogenic HPV types. Despite the high protective efficacy of these vaccines, not enough is known about the antibodies elicited. The aim of this project is to clone and characterize HPV-specific human monoclonal antibodies after vaccination, particularly antibodies reactive to types other than 16 and 18. Plasma cells and memory B cells were isolated from vaccinees at different time points following HPV vaccination. Antibody-encoding transcripts were amplified from pre-screened plasma cells and cloned into expression vectors - one for each antibody light and heavy chain. The expression vectors were co-transfected and the proteins were harvested and purified for one of the antibodies of interest. Antibodies reactive to HPV types 6, 11, 16, 18, 31,33 and 58 were identified. The first antibodies in the process of cloning were reactive to HPV type 11.The findings of this study will allow us to use these monoclonal antibodies as reference standards to determine the quantity of antibodies in serological assays, thus helping us identify the binding affinity and neutralizing capacity of vaccinees’ antibodies. This will aid future studies aimed at answering the question of whether the vaccine doses can be reduced to one, instead of two or three doses.

Can parenting styles offset the effects of neighborhood disadvantage on child life-course trajectories? Much research indicates that children raised in an economically disadvantaged neighborhood face a greater risk of high school dropout or expulsion, low-wage employment, and incarceration. Yet not all youths raised in disadvantaged neighborhoods follow this trajectory, and success stories abound. This study examines how low-income parents’ childrearing strategies positively or negatively affect the life-course trajectories of at-risk youth. Using the Denver Youth Survey (DYS), a nationally representative, longitudinal sample of high risk youth, I intend to test whether parents’ childrearing strategies affect young adults’ academic accomplishment, income, job type, and criminal record. I hypothesize that positive parenting, defined as monitoring, control, and warmth towards the child, is associated with a reduced likelihood of youths experiencing impediments to socio-economic mobility. As an alternative hypothesis, I test whether impulsivity of the child mediates the association between parenting style and positive life-course outcomes. In doing so, I aim to contribute to the intersecting literature on life-course theory, child developmental psychology, and criminology, as well as provide evidence to aid the development of policy assisting at-risk youths overcome their disadvantaged circumstances.

Although incarceration is ostensibly designed to punish people for breaking the law, it also punishes their families. The current work focuses on consequences resulting from parental incarceration. We examine the role this factor plays in peers’ moral cognition and pro-social behavior. Younger (5- to 6-year-old) and older (7- to 8-year-old) children rated the extent to which they viewed peers with or without an incarcerated parent as holding certain moral beliefs. Both groups of children were more certain that peers with, versus without, an incarcerated parent were less knowledgeable about morality. However, this effect was strongest among older children. This suggests that negativity toward children of incarcerated parents is socially reinforced and that children whose parents are imprisoned may face different peer responses depending on their age. Further, regardless of age, participants shared more resources with the peer whose parent was not incarcerated compared to the peer with an incarcerated parent. Taken together, these data clarify children’s moral cognition and pro-social behavior. They also shed light on the social realities that children of incarcerated parents may face.

The existence of 'checkpoints' in the mitotic cell cycle is well characterized; however, much less is known about the possible existence of checkpoints controlling re-entry of terminally differentiated cells into mitosis. We are exploring this possibility in Chlamydomonas reinhardtii, a unicellular eukaryotic green alga that, in the presence of visible light, differentiates back and forth from an actively dividing 'vegetative' state to a non-dividing 'gametic' state depending on the presence or absence of nitrogen in the culture medium. The work of previous students in our group suggests that gametes of wildtype C. reinhardtii show a growth delay of about 12 hours after nitrogen is added to nitrogen-deficient media; UV exposure immediately prior to nitrogen addition significantly increases the length of this delay. This is consistent with the working hypothesis of a dedifferentiation checkpoint in this organism. However, the visible light requirement for dedifferentiation confounds this result, because C. reinhardtii also uses visible light for photoreactivation subsequent to UV exposure. To control for this, we are taking a genetic approach. We expect to find an even longer post-UV dedifferentiation lag for a photorepair-deficient mutant of C. reinhardtii compared to wildtype, which would strengthen the case for a dedifferentiation checkpoint in this organism. Of particular interest in the present study is our use of two parallel statistical approaches to analyze our experimental data: (1) a 'traditional' timepoint-by-timepoint statistical analysis, and (2) statistical comparison of logistic curves fitted to the same data. We have written programs that facilitate execution and comparison of the outcomes of both methods of analysis, and expect both methods to agree in confirming our hypothesis. This insight into the inner workings of cell cycle dedifferentiation, as well as the validation of the second, novel statistical approach could be beneficial to a broad variety of ecological and biomedical experimental contexts.

Harnessing the power of enzymes to carry out synthetically relevant reactions is rapidly emerging as a powerful tool for sustainable chemistry. Iron-dependent enzymes have been of particular interest to the field because of their ability to facilitate complex reactions with broad substrate scope. Recently, we found that Fe(II) 2-oxoglutarate dependent hydroxylases (Fe(II)/2OGs) exhibit non-native activity towards either epoxidation or allylic hydroxylation. Oxyfunctionalizations are important in chemical synthesis as they allow for diversification to a range of more complex molecular scaffolds from simple olefinic precursors. Because of this, reactions that produce hydroxides or epoxides are of high interest, especially in an asymmetric fashion. However, current methods are not broadly applicable, as they are limited in substrate scope, selectivity, and tolerance towards other functional groups. Enzymes could rival traditional methods, offering greater selectivity and substrate scope while using inexpensive and Earth-abundant reagents. Here, we explore the ability for Fe(II)/2OGs to catalyze non-native asymmetric epoxidations or allylic hydroxylations. First, we will determine whether the reaction taking place is an epoxidation or allylic hydroxylation. Second, we will identify any additional Fe(II)/2OGs capable of oxyfunctionalization beyond our initial hit. Third, we will use directed enzyme evolution to improve oxyfunctionalization activity and enantioselectivity of a selected enzyme candidate. Fourth, we hope to expand substrate scope of an evolved Fe(II)/2OG to develop a general “epoxidase” or hydroxylase capable of reacting with a variety of functionalized olefinic small molecules in an asymmetric fashion. Generally, our lab seeks to use Fe(II)/2OGs mutagenesis libraries and a developed liquid chromatography-mass spectrometry screening platform to exploit the existing wide catalytic diversity of Fe/2OGs into a useful array of synthetically relevant transformations.

Jim Crow era legalized racism denied Black women the freedom to exercise control over their childbearing and childrearing; specifically, by restricting their access to necessary medical care and sufficient resources to care for their families, and by constraining their autonomy and agency. As a consequence, Black women experienced uniquely poor reproductive health and family outcomes compared to all other racial and ethnic groups (Eichelberger et al. 2016); these racial disparities persist today. This study applies a reproductive justice framework to understanding Black women’s lived experiences of systematic raced and gendered oppression, as well as their forms of resistance when caring for themselves and their children. Reproductive justice is the personal right to control one’s body, have children under the conditions that we choose, and parent those children in stable communities (Sister Song 1997). Thus, we ask how did gendered racism impact Black women’s experiences of reproductive justice and what strategies of resistance did they devise in response? We used Dedoose, a cloud-based mixed methods software, to conduct a content analysis of oral histories from two oral history repositories. These primary sources were excerpted and coded for common themes including racism’s influence on childbearing and childrearing, socioeconomic experiences, access to medical care, and protective factors. We have three preliminary findings that contribute to existing literature: 1) when women required more medical care than midwives could provide, they experienced numerous barriers to accessing such care, 2) Black women experienced multiple levels of social control that undermined their childrearing, and 3) women devised strategies of resistance to care for their bodies and their children, including collective childrearing, resource sharing, and instilling a sense of self-worth in their children.

Training a nonhuman primate (NHP) for research experiments generally requires the NHP to spend large quantities of time learning experimental tasks outside their home environment, and this requires a human researcher to be present at all times during training sessions. The purpose of this project is to create a wireless, semi-autonomous, low cost, cage-side training reward system allowing NHPs to train on experimental tasks for extended periods of time without the presence of a human researcher. An ideal device would allow for wireless data collection and provide both real-time and post-training information on the NHP’s training progress. Exposing NHPs to tasks first in the low-stress environment of their home cage before exposing them to the same task in an experimental booth can potentially speed up training processes. This lets research laboratories maximize researcher time and efficiently use equipment. Our cage-side training reward system consists of an iPad displaying touchscreen tasks, a speaker supplying audial cues for the tasks, an automatic feeder administering treats to the NHP for correct performance, and a computer to control the touchscreen tasks and collect data with custom MATLAB code. The iPad and computer communicate via a Wi-Fi router and this router also communicates with a Wi-Fi receiver which runs the feeder and speaker. The connection methods give the ability for wireless communication through walls, allowing the researcher to run tasks semi-autonomously from a computer outside the animal room. Excluding the costs of the iPad, computer, and MATLAB license, the system is estimated to cost under $300. Two rhesus macaques have undergone cage-side training with this device and have subsequently transitioned smoothly to learning tasks in a traditional experimental booth. In conclusion, this device serves as a low-cost method to enhance the training process for non-human primates while saving time and resources of the research laboratory.
 

 Loss of tooth mineral, demineralization, is the root cause of dental ailments - the most prevalent health problems affecting over 90 percent of Americans. These range from white spot lesions, the earliest sign of dental demineralization, to periodontal diseases, which can lead to more serious health issues. Current restorative treatments of tooth structure and function utilize synthetic materials, e.g. amalgam, glass ionomers, and particle reinforced resin composites that lead to deposited secondary precipitates. Although these common procedures are well-established and relatively effective, their durability is limited due to lack of structural and functional integration of deposited layer with the underlying tooth. GEMSEC labs have developed a proprietary technology dubbed “peptide-guided remineralization” which enables the formation of a new mineral with protein-derived peptides. Using this technology, the lab teams have successfully restored dental hard tissues via several case studies including enamel, cementum, dentin under in vitro and in vivo conditions. Translating this technology into a daily-use product, we developed a prototype, dental lozenges, designed to aid in enamel remineralization using a biomineralizing peptide, ADP5, derived from amelogenin, the key enamel protein. Herein we aim to refine the lozenge formulation through an iterative study for enhanced durable and whitening remineralized layer. Remineralization performance of different tablet formulations were tested in artificial saliva using extracted human enamel teeth. The samples were characterized using SEM showing that the current lozenge formulation creates a new mineral layer on enamel up to 2 µm in thickness. In summary, the new lozenge artificially regenerates lost enamel on the molecular level to treat tooth decay and erosion. Developed through a simple biomimetic methodology, this prototype lozenge could be mass fabricated for the consumer dental care market and expanded to include dental varnishes, gels, and pastes.

Thalassinidean shrimp, commonly known as mud and ghost shrimp, are derailing oyster beds, a profitable aquaculture venture in Washington, by burrowing under them and causing sinking. In an attempt to replace harsh chemicals as pest control, a brine solution to osmotically eliminate these shrimp has been researched. Osmoconformers cannot control their internal pressure and are susceptible because of high gill surface area. Unfortunately, when this high salinity water is pulled back to the ocean at high tide, subtidal invertebrates, like the common shrimp Pandalus danae are also potentially impacted. Here we show the death rates of P. danae in varying salinities, and gill dehydration due to osmotic pressure through histology. Mortality rates were high for all salinity treatments. All the gills processed through histology showed intense dehydration. Though time of death, and thus time of dehydration, varied by salinity treatment, amount of dehydration did not. Runoff from the brine treatments to kill mud shrimp will impact other species in the subtidal, especially other osmoconformers. They have no way to retain water and thus are very susceptible. This study could provide information about ecological impacts of other animals in the brine solution beyond just killing the mud shrimp.

Globally, wetlands provide important ecosystem services and are critical to supporting wildlife and biodiversity. Anthropogenic disturbances, such as road construction, have a negative impact on wetland health and have dramatically reduced their number worldwide. In response to the damage caused by road construction, the Washington State Department of Transportation (WSDOT) mitigates the consequent reduction of functions and the loss of wetlands through restoration efforts, including the monitoring and eradication of invasive vegetation (e.g. reed canary grass). WSDOT currently maps and monitors invasive species on the ground, which is challenging as they are hard to access due to inundation and dense vegetation. Compared to field survey methods, drones have the potential to quickly and safely survey large areas, reducing human effort and cost. By focusing on a single mitigation wetland site, we investigate the use of drones as an effective tool to accurately survey reed canary grass. We use object-based image analysis (OBIA) to create maps of reed canary grass cover and test the accuracy of the map using visual interpretation and confusion matrices. Results will inform about the difference in map accuracy between three drone sensors, an add-on 5-band (red, green, blue, red-edge, near-infrared (NIR)) camera and two built-in 3-band (reed, green, blue) cameras. We discuss opportunities and limitations of using drones as a tool to map invasive species. Additionally, we highlight the considerations that ecologists and natural resource managers must take into account when using drones for wetland monitoring. In conclusion, we identify future areas of research that include testing the repeatability of these methods at additional wetlands and increasing the suitability, number, and timing of the field data in support of this work.

Osteoporosis is an orthopedic disease in which old bone begins to dissolve but is not replaced by new bone. This reduces overall bone density and increases a patient’s risk for fractures. One human gene associated with osteoporosis-related traits is WNT16, which is also expressed in zebrafish. Previous studies in our lab have shown that wnt16 mutant fish have skeletal defects. The goal of this project was to find the most effective method for knocking out genes associated with osteoporosis in zebrafish using CRISPR/Cas9 technology. Once Cas9 creates a double-strand break in the DNA, there are different methods of DNA repair, two of which are non-homologous end joining (NHEJ) and microhomology mediated end joining (MMEJ). In this study, we designed guide RNAs (gRNAs) targeting wnt16 to compare a more predictable, MMEJ-based CRISPR approach to the previous, NHEJ-based CRISPR approach used in the lab. Next, we assessed gene editing in zebrafish embryos injected with two new gRNAs biased toward MMEJ repair to determine their efficacy in knocking out wnt16. Using DNA sequencing and analysis, we found that injections of both MMEJ gRNAs caused high rates of insertions and deletions (indels) compared to a control group which was not injected with gRNA. Moreover, the predicted MMEJ indels were found to be in high abundance in DNA sequences from injected fish. Further, we detected anticipated morphological differences expected from loss of wnt16 in the fish injected with MMEJ gRNAs compared to the control fish, suggesting that the MMEJ gRNAs work as expected. Based on these results, MMEJ-biased gRNA design appears to be a promising approach to improve efficiency in knocking out zebrafish genes. This project may help optimize a rapid and effective screening of many zebrafish candidate genes using CRISPR/Cas9 technology to study skeletal phenotypes in zebrafish.

Euler integration is the simplest, most versatile, and underappreciated method of integrating partial differential equations (PDEs) that only involves repeated addition. Evaluating the evolution of connected dynamical systems is critical to fundamental research as well as to students’ understanding of the physical world and their classwork. The purpose of this research is to design and implement an educational tool that empowers students and faculty to understand the beautiful simplicity of the most applicable method of evaluating PDEs on a computer. Physical law is always written in the form of a PDE. Traditional physics education does not emphasize this technique. This is largely due to the lack of computers over the last six hundred years. But now, we have super computational ability at our fingertips, and it is time that everyone in the field of physics has access to this versatile and simple tool. The first educational tool is complete and has already helped students learn about this technique. Over the next few months this tool and ones like it will be sewn into existing curricula in the physics department. This technique applies to an enormous range of disciplines from fungal growth to fluid dynamics and will be a skill at every student’s disposal.

Fear is an emotion that is triggered when an organism encounters danger or threat. Once fear is triggered, the nervous system induces many physical changes, and the brain responds accordingly to elicit defensive or fleeing behavior. Such fear responses are known to involve a critical brain region called the amygdala. One of the ways in which the amygdala mediates fear response is by secreting a neuropeptide called corticotropin-releasing factor (CRF). Our lab has previously found that CRF-expressing neurons in the central nucleus of the amygdala (CeA-CRF neurons) facilitate the acquisition of discriminatory fear responses and prevent fear extinction. In contrast, others have reported that activation of CeA-CRF neurons is reinforcing. To resolve these contradictory findings, we investigated the functional roles of CeA-CRF neurons in naïve and fear-conditioned mice. We hypothesized that activation of CeA-CRF neurons in a neutral context may produce reinforcing effects by signaling a type of salience. However, following a fearful experience these neurons may signal the aversive nature of this experience. To stimulate CeA-CRF neurons selectively, we used optogenetics, in which light-sensitive ion channel-expressing neurons are stimulated using light, and animals underwent several behavioral tasks including operant conditioning for lever pressing, real time place preference task, and elevated plus maze test. Based on the animals’ emotional context (naïve or fear-conditioned), we examined how stimulation of CeA-CRF neurons affected the animals’ behavioral performances. Our results showed that stimulating CeA-CRF in naïve mice produced no effects. However, if the animal’s first experience in a context was appetitive, then stimulating in this context is reinforcing. Similarly, if an animal’s experience is aversive, then stimulating these neurons perpetuates fear and anxiety behaviors. These results suggest that CeA-CRF neurons signal context-dependent behavioral state effects that may be important for engaging appropriate actions in specific contexts.

Cocaine addiction is a health concern globally. Although both men and women get addicted to cocaine, women transition from casual drug use to addition faster than men, have greater difficulty remaining abstinent and demonstrate greater propensity to relapse. In our laboratory, we found that cocaine craving is potentiated in female rats specifically in the estrus phase of the hormonal cycle. The lateral habenula (LHb), an epithalamic nucleus, plays an important role in cocaine taking and seeking and also mediates several estrogen-dependent behaviors in female rats. In the brain, the effects of estrogen are primarily mediated by estrogen receptors alpha and beta. The goals of this study are - a) to determine if there are hormone-dependent changes in estrogen receptor plasticity in LHb neurons and, b) to define the neuroanatomical organization of estrogen receptors in LHb projection neurons. Firstly, brains from freely-cycling female rats were collected during all phases of the hormonal cycle and analyzed for LHb estrogen receptor expression. Preliminary results indicate that majority of estrogen receptor alpha expressing neurons are located in the ventromedial aspect of the medial to caudal LHb. Analyses are currently underway to determine if cyclical fluctuations in ovarian hormones across the estrus cycle influence estrogen receptor expression in LHb neurons. Secondly, rats were injected with canine adenovirus (CAV2) expressing ZsGreen into either the ventral tegmental area (VTA), dorsal raphe nucleus (DRN) or rostromedial tegmental nucleus (RMTg), three LHb targets. CAV2-ZsGreen vector was retrogradely transported to neuronal cell bodies in the LHb where ZsGreen transgene was expressed. RNAScope in situ hybridization studies are in progress to determine the co-localization of ZsGreen positive neurons in the LHb with estrogen receptors. Understanding the cyclicity of LHb estrogen receptor expression will guide our studies targeted at understanding sex differences in neuronal mechanisms of cocaine taking and seeking behaviors.

Nearly half of Americans live in earthquake prone areas. Many primary fault zones that host large earthquakes, such as the Cascadia, Alaska, and San Andreas fault zones, extend into the offshore regime. These offshore fault systems have been historically difficult to study due to challenges in observational techniques. Through the creation of an algorithm that uses geospatial analytical tools, this study seeks to identify seafloor faulting structures from data collected by high frequency multichannel acoustic methods. In doing so, we improve our capabilities of characterizing offshore fault zones. In addition, we examine these geospatial analytical methods for accuracy and explore the impact of data collection and post-processing procedures on associated errors. Data utilized subsists of bathymetric data collected in the Cascadia and South African regions, which are active and passive margins respectively. Methods for surface fault identification include visual inspection, as well as geospatial analytical methods consisting of the Bathymetric Position Index, slope, and aspect of surface morphology. Faults identified from surface morphology are compared to those identified using a coherence-based detection method from seismic reflection data. Surface expressed faults indicate high-amplitude and/or recent geologic deformation and can give insight into tectonic stress regimes and associated faulting hazards. An improved understanding of faulting hazards through efficient surface fault identification would aid in preparation and planning for earthquakes. Through the creation of this algorithm, our capabilities to accurately identify surface expressed faults in bathymetric datasets will be enhanced and thus our understanding of global tectonic processes and earthquake risks to population centers like those in the Pacific Northwest will be improved.

An important issue in addressing the cocaine addiction problem is the tendency and probability of recovering addicts to relapse to cocaine after a significant period of time not-using. External cues that have been conditioned to be associated with cocaine use in the past are powerful triggers that cause relapse to cocaine-seeking behaviors. Drug relapse is often caused by experiencing “craving,” a subjective affective state that motivates one to seek drugs. Cocaine craving increasing as a function of time has been shown in the laboratory setting in rats and subsequently dubbed as the “incubation of cocaine craving.” This phenomenon shows that craving increases as a function of time in response to these conditioned cues. Although both women and men have a path to addiction with cocaine, women experience a faster transition from casual use to drug addiction, have a harder time remaining drug-free, and display a greater probability of relapse compared to men. Pre-clinical studies showed that female rats demonstrate an enhanced incubation response when re-exposed to conditioned cocaine cues and display conditioned place preference at lower doses of cocaine than male rats. However, the brain regions that underlie the enhanced incubation response are unknown. We trained cycling female and male rats in the laboratory to self-administer cocaine on a continuous long-access (6h), fixed-ratio 1 reinforcement schedule (reinforcement administered after one correct response) for 10-14 days. After self-administration training, rats were exposed to the cocaine self-administration environment on abstinence days 1 and 35. Our aim for this experiment is to investigate the brain regions that underlie incubation. We are quantifying the immediate early gene c-Fos immunoreactivity in cells across various brain regions in order to determine the similarities and differences in the neural activation during incubation of cocaine craving in female and male rats. We expect to see increased c-Fos expression in animals that are incubuating craving.

A graph is a collection of vertices and edges. In computer science, graphs are often called networks and form the basis for many data structures and search algorithms. A matching of a graph is a selection of edges that share no common endpoints. The set of all matchings of a graph forms a simplicial complex which we call the matching complex. We are interested in the relationship between a graph and its matching complex and have been exploring whether we can characterize all simplicial complexes that are matching complexes. What structure does the matching complex imply about the graph and vice versa? We have also been interested in connections between matching complexes and well-known simplicial complexes– in particular, two-dimensional Buchsbaum complexes. These have much more structure than simplicial complexes in general, so they lend themselves to interesting questions. How can we characterize all two-dimensional Buchsbaum complexes that are matching complexes? We have also developed an interest in sequences of graphs generated by taking repeated matching complexes. Understanding these sequences would allow us to categorize graphs using the matching operation. Which graphs go to the empty set after a finite number of iterated matchings? For what finite values does this occur? What common structure do these graphs have? Investigating these questions will allow us to categorize graphs and complexes using the matching operation. We hope to make connections between graphs or categories of graphs that would otherwise remain disconnected. In pursuing these queries not only are we seeking answers but a development of tools which can be applied to further exploration.

Every minute, Twitter users send hundreds of thousands of tweets, providing a rich resource of publicly available text data. Our goal is to use this data to learn from and imitate the sentence structure of specific accounts. To this end, we develop mRkov, a statistical tool that takes the username of a Twitter account, also known as a handle, as input and outputs fake tweets that mimic the linguistic style of the tweets from that handle. We built mRkov into an R software package as well as an interactive and user-friendly web tool that walks the user through the process of using our software. mRkov first scrapes tweets posted from the input Twitter handle, and then after processing the text and sentiments of the scraped tweets, generates new tweets using Markov chain simulation. Markov chains consist of a sequence of items, where each item is probabilistically sampled dependent only on the preceding item in the chain. By using non-independent sampling, the Markov chain method generates a sample that mimics the true distribution. In this application, the Markov chain is a sequence of words, and the distribution is the sentence structure of the tweets. mRkov also allows users to provide input that influences the sentiment of tweets in order to generate tweets that tend to be more “positive” or “negative” in sentiment. Tools such as mRkov help us better understand patterns of speech and writing. This has many useful applications, including identifying if multiple accounts are coming from the same source or writer, analyzing and comparing how the style and sentiment of different accounts change over time, and detecting bots or other fake accounts.

Ongoing research has been conducted to design optimal routes for vaccine distribution to health centers in Mozambique. Distance between health centers is needed in order to construct routes, but is not easily available. My objective is to easily generate distances between health centers, given their geographic coordinates, and to visualize the optimal routes on a map. Using a conglomeration of tools including Python, API, OpenRouteService and Excel, I was able to read an Excel file of latitudes and longitudes for health centers, separated into provinces and districts, and produce an Excel matrix of distances from health center to health center as well as an API map visualizing the routes to and from the given health centers. My Python code utilizes OpenRouteService with an API that provides data including distance, travel time, direction, maps, etc. OpenRouteService also allows the user to specify their route preferences. We can choose the road type, the speed limit, the time of travel, the mode of transportation, etc. The created distance matrix and map will be used in an interactive route optimization tool. The route optimization tool allows end users to efficiently distribute vaccines using available vehicles. The distance matrix gives the tool correct distances between health centers and the map gives visualizations of the distances. The optimization tool provides routes for efficient distribution and my Python code maps the routes for easy interpretation of the delivery system. Moving forward, we want to make the code more user-friendly, so anyone can create a new and improved version.

Water is the substance that exists everywhere in our lives ranging from drinking water to the blood in the body. It is well known that there are 3 phases of water: gas, liquid and solid. Yet, in Dr. Gerald Pollack’s lab, we conduct researches on the Exclusion Zone (EZ) water, which we term as the ‘Fourth Phase of Water.’ Dr. Pollack’s lab centers largely on the identification of EZ water and many applications in nature and technology. Among the natural applications, the lab emphasizes on the role of EZ water in human health, including cell biology since cells are filled with EZ water and cannot function without enough EZ water. Dr. Pollack’s lab conducts the research uncovering the nature’s hidden secret that has tremendous potentials to be applied to different bioengineering products. I am currently conducting a project on how Wi-Fi impacts EZ water as an external source of disturbances. Humans are exposed to Wi-Fi signals constantly in our everyday lives. As human body cells are filled with EZ water, we predict the Wi-Fi signals could alter our bodily functions through changes in EZ water properties such as amount of EZ water. EZ water develops around hydrophilic substances, and we uesed blood vessel-like tube to observe EZ water development. Then, we measured the amount of EZ water and analyzed the data. For this project, we compare the difference in the amount of EZ water built with and without presence of Wi-Fi. We currently have statistically significant results that Wi-Fi decreases amount of EZ water developed by ~18%. As a further step in the future, we are investigating on how different types of disturbances such as cell phone impact on EZ water for further health care.

Prior research has established that the occurrence of headache in adolescence is more likely if one or more parents have a headache history. However, little is known about the prevalence of non-headache (i.e. musculoskeletal or abdominal) pain history among parents of adolescents with headaches, and whether adolescent headache characteristics (frequency, pain intensity, functional disability) differ among different parental pain histories. This analysis aimed to characterize the parental pain history of adolescents with recurrent migraine and tension-type headaches and examine if differences in parental pain history predicted different adolescent headache characteristics. 238 adolescents (mean age = 14.64 years (SD = 1.86), 67% female) who reported at least 10 headaches days per month completed a 28-day headache diary and a self-report questionnaire on headache-related disability. Parents (mean age = 44.06 years, SD = 6.04; 94% mothers) completed a pain history checklist. Results indicated that most parents had a positive pain history (78%). Parental musculoskeletal pain (55%) was just as prevalent as parental headache (54%; χ²(N = 226) = .098, p = .754), but abdominal pain (27%) was significantly less common (Headache: χ²(N = 227) = 30.306, p < .001; Musculoskeletal pain: χ²(N = 226) = 44.298, p < .001). Most parents reported pain at one site (35%), followed by two sites (29%), having no pain (23%), and pain at all three sites (14%). Adolescent headache characteristics were similar between parents with or without a pain history and when parents had a headache and/or musculoskeletal pain history, and were not associated with the number of parental pain sites (ps > .05). These findings indicate that any connection between parental and adolescent pain characteristics may be more complicated than a simple direct relationship. Future research should examine whether the characteristics of parental pain (e.g., parental pain intensity) are associated with headache characteristics in adolescents.

Adolescence alcohol use and opioid addiction in adults are systemic issues afflicting the world, and thus, it is important to elucidate the long-term individual and relational consequences of both substance abuse disorders. The purpose of this experiment was to examine the long-term consequences of voluntary adolescent alcohol use on morphine tolerance, fentanyl self-administration, and the effects of previous opioid exposure on fentanyl self-administration in adulthood. Using a preclinical model to examine this hypothesis, adolescent rats had access to alcohol in gelatin form for twenty days, after which a three week withdrawal period occured. Morphine was then administered intraperitoneally for five days and morphine tolerance was measured by a tail-flick test for those five days. Finally, fentanyl self-administration occured in an operant chamber and self-administration will be measure by the amount of fentanyl consumed. My expected results were that adolescent alcohol use will increase morphine tolerance as evidenced by decreased tail-flick time, and fentanyl self-administration will also be increased. Additionally, I expect that previous opioid exposure will increase fentanyl self-administration. Future research should examine the neurobiological mechanisms by which adolescent alcohol use increases morphine tolerance and fentanyl self-administration and how previous opioid exposure increases fentanyl self-administration since these biological mechanism are not well understood.

 The number of older adults ≥65 years with bothersome pain has increased in the past decade due to an increase in population size and life expectancy of older adults. Amongst several factors studied that may contribute to pain prevalence, socioeconomic status(SES) has been concluded in most studies to be more directly associated with pain prevalence rather than race/ethnicity. The objective of this project was to determine how physical function, due to pain, varies by race/ethnicity. We conducted a retrospective secondary analysis on the National Health & Trends Study(NHATS) to determine how Short Physical Performance Battery (SPPB) scores and secondary pain outcomes differed by race/ethnicity amongst participants who reported bothersome pain in 2011. Baseline race/ethnicity characteristics were gathered to characterize cohort. SPPB score and secondary outcomes for white(NH), black(NH), and hispanics were described for Rounds 1&2. Weighted, regression analysis were performed on SPPB scores to adjust for covariates using survey features. Results show a meaningful difference of SPPB scores among racial/ethnic groups that prompts for more research as to what factors may play a role in differences on the impact of pain on physical function by race/ethnicity.

Diets high in fat and sugar have increasingly adverse effects with increasing age because of generally decreasing activity and energy needs. This nutritional overload results in an increase in body fat mass and an increase in risk for metabolic and cardiovascular disease and possibly other chronic diseases such as cancer. However, the role of fat mass in age-related pathology and survival over an extended period of time is controversial. To address this issue, C57BL/6 mice, 18 months of age, were started on a high caloric (HC) diet consisting of balanced protein, lard, and sucrose. A second cohort was maintained on a standard rodent caloric (RC) diet consisting of balanced protein, wheat, corn, and soybean oil. Fat mass was measured by quantitative magnetic resonance imaging (QMRI) monthly for 10 months (28 months of age) at which time mice were evaluated for physiological performance, and tissues collected for geropathology. Both diet groups started with an average 5 percent fat mass. Fat mass increased to 15 percent in the HC diet group over the next 3-5 months, then gradually decreased to 9 percent after 10 months on the diet. Fat mass gradually decreased to 3 percent in the RC diet group over 10 months. Mice fed the RC diet had increased paw grip strength and were able to stay on a rotating rod longer than mice fed the HC diet, but there was no difference in survival between the two diet cohorts over the 10-month trial period. These preliminary observations suggest that percent body fat mass generated by a diet high in animal fat and table sugar may be associated with unhealthy aging, but not survival. This provides rationale for subsequent investigations into the pathological consequences of high caloric diets with increasing age.

Marine heatwaves are the phenomena of abnormally warm ocean surface temperatures that last for an extended period of time. The most severe marine heatwave of recent times occurred from 2013 to 2016 in the Northeastern Pacific. This event, nicknamed ‘The Blob’, was scientifically fascinating because the ocean-atmosphere system maintained itself for so long in an anomalous state. In mid-2019, a marine heatwave with a likeness to ‘The Blob’ began forming. This research project focuses on analyzing the anomalous patterns in sea surface temperature, clouds, radiative fluxes, and turbulent fluxes that arise during the formation and duration of this event. We set out to understand if the more recent 2019 marine heatwave evolves in a similar way to that of ‘The Blob,’ and how it differs. This project uses NOAA Climate Forecast System Reanalysis (CFSR) data, which assimilates measurements using complex models to create the best estimates of atmospheric and oceanic variables with complete global spatial coverage. With this project, we aim to understand the atmospheric response to marine heatwaves using geospatial plots of mean temperature, fluxes, and cloud cover. We expect to see differences in the atmosphere response with regards to the net flux that caused the quick dissipation of the recent marine heatwave.