Student Research Presenters

Rolling from a supine to prone position marks an important developmental milestone for infants. While previous studies have recorded infant rolling patterns, these are conducted in complex laboratory settings with limitations. Research laboratories are an unfamiliar environment for infants which may not simulate how infants roll at home. Identifying these rolling patterns also requires expensive motion capture equipment which is not readily available for all researchers or clinicians. Accurately classifying infant rolling patterns throughout development is crucial in the early diagnosis of possible developmental delays or disorders. Therefore, using video techniques could be a promising approach to study infant rolling movements throughout development that is readily available for both researchers and clinicians. In this study, we aim to present a methodology that allows researchers to accurately and consistently categorize infant rolling patterns via at-home video analysis. The methodology consists of three steps: 1) identifying roll direction, 2) identifying stationary and moving limbs, and 3) determining synchronicity of moving limbs. Detailed descriptions and illustrations of each coordinated movement were presented to aid the viewer in categorizing videos into six different movement types. Three reviewers were tasked with categorizing 45 videos of infants achieving a roll using the methodology. Fleiss’ Kappa statistical analysis was used to evaluate inter- and intra-rater reliability. The overall inter-rater reliability score was 0.694 and the overall intra-rater reliability score was 0.801, both classifying as substantial agreement. These results suggest that this methodology can produce consistent results when evaluating infant roll patterns through video. 

Scientists have discovered that prolonged exposure to microgravity leads to degenerative changes in both bone and muscle tissue, but the precise mechanism behind this remains unknown. It is known, however, that the LINC complex pathway plays a crucial role in recognizing mechanical input and transmitting signals to stem cells to produce different tissue types. It is the absence of these signals that causes these degenerative changes to occur in microgravity conditions. Previous research on mesenchymal stem cell (MSC) nuclei and their nanomechanical properties showed changes in cellular stiffness as part of the mechanotransduction pathway. Using atomic force microscopy (AFM) cantilever-based nanoindentation, this technique measures the cellular stiffness of the nuclei and provides insights into the effects of different interventions, such as low-intensity vibrations, genetic manipulations, and pharmaceutical treatments on the stem cells applied by collaborators, Dr. Gunes Uzer and Dr. Sean Howard. By studying cell nanomechanics and nanoindentation, we aim to determine the impact of mechanotransduction on cell stiffness and identify potential pathways for mitigating the negative effects of microgravity on human health during extended space travel.

This study has revealed that, although minority students have a sense of belonging in identity-based organizations, they do not feel the same in the classroom environment or with STEM faculty. Identity-based resources in universities can be a great way to represent and support minority students [1]. However, resources should also follow through to classroom environment. Faculty should be aware of the racial inequity between minority students and nonminority students. As important members of society, faculty have the responsibility to inspire and drive generations to succeed. Faculty should strive to close this racial inequity in the classroom and through their teaching style. This study focused on understanding if and how minority students in STEM “feel a sense belonging” at Boise State University [2]. We conducted 14 individual interviews with minority students who are a part of the Louis Stokes Alliance for Minority Participation (LSAMP) program. Together, we analyzed and discussed the interviews through the use of a collaborative empathy map. We found that minority students felt a sense of belonging, had family-like relationships, and were comfortable in their own skin when at identity-based organizations. Although, when it came to faculty and classroom environments students did not feel the same. We found that minority students didn’t build meaningful relationships with faculty, faculty does not reach out to students, and classroom environments often alienate them. We recommend that (1) BSU faculty gain understanding of how differently minority students’ experiences and feelings are towards classroom vs. identity-based programs, (2) BSU faculty should learn from the identity-based programs and implement successful components in the classroom (3) BSU faculty should create a safe space in the classroom that allows for students to comfortably talk and participate in discussion.

References

[1] E. Knaphus-Soran, D. Hiramori, and E. Litzler, "Anti-racist institutional transformation matters: how can community cultural wealth and counterspace processes illuminate areas for change?," in American Society for Engineering Education Annual Conference and Exposition, Online, 2021.

[2] T. L. Strayhorn, College students' sense of belonging: A key to educational success for all students. New York, NY: Routledge, 2012.

My research will focus on the Student Media Department at Boise State University – a student-run organization aimed at producing hyper-local media for the student newspaper, university radio station, and an independent magazine. Through ethnographic methods, such as contextual interviews and referencing physical artifacts, I will produce a qualitative analysis of how Student Media employees actively gain professional experience. My study will encourage participants to reflect on their personal and career goals within the organization, while also informing the department how they can best uplift students transitioning into professional environments.

As younger generations increasingly prioritize meaningful work in their careers, the results of this research will assist in documenting the social contexts shaping a new workforce of creative young adults. While collecting data, it became evident that the department has provided a level of autonomy such that an employee’s personal interests can inform their education and career path. The functionality of the department is dependent on self-directed motivation – which largely reflects expectations within “real-world” professional environments. Across the 3 organizations, a variety of media content is published daily, leading to extensive portfolios reflecting individual skill sets. Their work is often revised by peers in a collaborative process, requiring them to receive and utilize relevant feedback. When asked about sentimental physical artifacts related to their jobs, it revealed a level of intellectual maturity that wouldn’t be possible for somebody who didn’t have to navigate their opinions so formally and on such a public scale – it was a roadmap for what influences their portfolio.

The analysis also raised valid challenges of the department, including various barriers to recruitment and limitations within university environments; this helped to inform the new-hire process, departmental procedures that need more attention, and improved planning to best support student employees. It is apparent that the Student Media Department acts as a springboard for creative young adults to truly bolster their passions by means of professional development, coursework connections, and portfolio curation.

Air pollution, specifically PM2.5, is linked to adverse health effects including premature mortality, pulmonary inflation, accelerated atherosclerosis, and altered cardiac functions (Nowak et al., 2010). Air pollution has also been shown to contribute to health disparities for low-income, and BIPOC people (Rigolon et al., 2021). The objective of this study is to determine if a correlation exists between proximity to greenspaces and PM2.5 concentration in South Seattle. A correlation coefficient and p-value were calculated using publicly available purple air data from June 1, 2022, to September 30, 2022, to find the average PM2.5 concentration at 6 meters within a 4 km radius of the Cheasty Greenspace in South Seattle. Using an online correlation calculator, the distance from the meter to the greenspace and Average PM2.5 concentration was determined to be positively correlated with a correlation coefficient of 0.413 and p-value of 0.3929 which suggests that though PM2.5 levels do increase with increased distance from a greenspace the levels themselves are not significantly different. These findings indicate a need for increased green space in South Seattle to protect urban air quality and the health of those living in the area.

Watersheds provide a variety of ecosystems that are beneficial to both humans, plants, and animals alike. Therefore, monitoring ion concentrations in its waters that affect its ability to support these ecosystems are paramount. The purpose of this study is to deduce the differences and source of ion concentrations in paving over a creek vs leaving it unpaved in an urban setting. Two waterways on opposite sides of a populated area in the Thornton Creek Watershed were selected: North Fork which is largely day lit or unpaved and Littlebrook which is largely paved over. 3 sites of each, and one at their confluence were sampled using a syringe and a .25 micron sterile filter and were then run through a Metrohm 930 Compact Flex IC. Pavement samples in direct contact with the creek, or of similar composition of pavement in direct contact with the creek were soaked in ultrapure water and ran through the same IC device for evidence on ion leaching after 4 days of soaking and then 17. Ion concentrations of chlorides, phosphates, and nitrates were higher in North Fork and nitrite concentrations were found to be higher in Littlebrook. The pavement samples also showed indication of some ion leaching, and increased leaching over time. The higher chloride, phosphate, and nitrate concentrations in North Fork could be attributed to it being unpaved thus having higher surface area in contact with the air, proximity  to roads, heavy traffic, exhaust emissions, and a higher population. The pavement samples showed some indication of ion leaching and because Littlebrook has greater surface area in contact with pavement compared to North Fork this may contribute to higher nitrite concentrations. Results are inconclusive as to where the ions in the pavement are from and further testing is needed. Although ion concentrations in both waterways are considered safe levels these potential sources of ions and ion differences should be noted and kept track of to prevent harming the ecosystem of the Thornton Creek watershed.

Thornton Creek has a drainage area of 12 square miles in northeast Seattle. Due to its location, it is susceptible to pollution from water runoff that comes from surrounding roads. Historically, Seattle has used various road de-icing compounds, such as calcium chloride in a soy-base, sand, and eventually sodium chloride after a 2008 snowstorm that shut down the city. During the 1990s, various habitat restoration projects took place, and the surveys taken to measure project efficacy discovered that coho salmon were dying before they could make it back to their natal streams to spawn. Researchers studied this occurrence and ultimately attributed salmon mortality to toxic runoff from urban land usage. This study aims to investigate the effects of using sodium chloride to de-ice roads on salmon livelihood. Water samples were collected two to three times daily for a week from Thornton Creek during Seattle’s ice storm in December 2022. Then, sodium chloride concentrations were tested using a Metrohm 930 Compact Flex IC in February 2023. The results from these tests were compared to the EPA’s sodium chloride toxicity levels, and none of the samples exceeded the chronically contaminated sodium chloride concentrations for a single event. Although the results from these samples do not indicate chronic contamination, further research is necessary to determine the long-term effects of using sodium chloride to de-ice roads. Through continuous monitoring of sodium chloride concentrations, the findings of this study can aid in finding alternatives to road salt that have less of an environmental impact to maintain water quality and improve the livelihood of salmon that spawn locally in Seattle.

Research on pathogen intervention and ubiquitin modification has increased over the last few years but a lot is still unknown about ubiquitin-like modifiers (Ubl) and how or if they are modified by pathogens in the same way as Ubiquitin. In this work, two different studies conducted over the course of 2 years are highlighted. The first study focused on purifying and synthesizing four activity based probes in order to study enzymatic activity in the Ubl-modification systems. These probes were reacted with virulence factors isolated from different bacteria and it was concluded that there were unexpected bands formed between some of the probes and the bacterial proteins. In the second study, a SUMOreal assay was developed to investigate if bacteria regulate host SUMO signaling. Past pathogen research has shown that pathogens have ways of hijacking ubiquitin systems with their unique ligases, eliminating unwanted cellular proteins, and creating systems where they are able to control the environment for their own benefit. Both these studies helped with better understanding the roles of these modifiers in bacterial infection in order to ultimately understand crucial mechanisms of disease and aid in the elucidation of fundamental cellular biology. 

With an ever-changing environment threatening plant species persistence, broadening our understanding of selective mechanisms and their role in climate adaptation is more crucial now than it has ever been. In particular, there is a lack of genomic studies examining two selective mechanisms that occur during the reproduction of angiosperms. These selective mechanisms are referred to as gametophytic selection (GS), in which pollen grains compete to fertilize ovules, and selective embryo abortion (SEA), in which developing embryos compete for maternal resources. Analyzing deviations from Mendelian segregation offers an avenue for identifying loci targeted by GS and SEA, and their contributions to purging of genetic load. We have developed models of expected segregation patterns for GS and SEA, but other selective processes, including female meiotic drive, ovule abortion, and early seedling mortality, can mimic these expected patterns.  To distinguish the effects of GS and SEA from these other selective processes, we will conduct reciprocal crosses between highly homozygous and highly heterozygous genets of Mimulus guttatus. Segregation distortion at the pollen, seed, and F1 seedling stages will be analyzed for each direction of the reciprocal crosses to infer the mode of selection. We hypothesize that a proportion of the observed segregation distortion will be due to meiotic drive, which can then be accounted for when estimating selection due to GS and SEA. This work will reveal the relative contributions of GS and SEA to allele frequency changes that may ultimately affect the speed and trajectory of plant evolution.

The SARS-CoV2 genome encodes for its own highly conserved helicase, nsp13 which is crucial for replication of the virus. Understanding nsp13’s kinetic behavior could lead to potential ways of inhibition thereby preventing replication of the virus. Here using nanopore technology we observed that nsp13 translocated at different speeds depending on the sequence of DNA it walked on. This project aims to further investigate the sequence dependent kinetics of nsp13 using nanopore tweezers.

In a nanopore experiment a salt solution divided into two wells is separated by a thin membrane. In the membrane, there is a single nanopore. A voltage applied across the membrane causes an ion current through the nanopore which is measured. As negatively charged DNA is dragged through the pore, it blocks the pore differently depending on the sequence allowing us to determine unique currents for individual bases. A helicase, nsp13, translocates along the DNA simultaneously lowering it through the pore, allowing for observation of the enzyme’s kinetics. In our experiment we take a previously sequenced DNA template and replace a Guanine with a Cytosine and compare differences in how long nsp13 translocated on each base (dwell time).

We found further evidence that nsp13 does have different dwell times depending on what base it is translocating on and more specifically in the position where Guanine was mutated to Cytosine, a longer dwell time followed by smaller dwell time was observed. This data hints at an important location in the sequence, however more data is needed to fully understand how this affects nsp13 kinetics. 

Nanopore technology can provide significant information regarding enzyme kinetics, such as sequence dependence kinetics that could help us understand nsp13’s role in vivo. Nsp13 sequence dependency could be further applicable as it could be used to discover a way to possibly inhibit SARS-CoV2 replication. 

As composite materials are increasingly used in the aerospace industry, there is an ever increasing demand to develop processes to recover the waste generated. Typically, the thermoset composites such as carbon fiber pre-impregnated epoxy material can exceed its shelf and freezer life if not used within the allotted time specified by the material manufacturer. The current processes for recovering the expired carbon fiber prepreg either leaves the material irreparable during resin separation or requires a vast amount of energy to burn-off the resin. The benefits of designing a desirable approach to fully reutilize expired carbon fiber prepreg would extend across multiple industries. This would result in lower energy requirements to repurpose the material. Our current research focuses on the ability to recover the mechanical properties of expired aerospace grade prepreg by reverting the unidirectional material into fiber form. To fully disperse the fibers, the material is added to a solvent that best fits the solubility parameters of the resin in the prepreg. The solvent and prepreg are agitated by an overhead stirrer that can produce enough shear force to aid in the dispersion of fibers. Once the desired homogenized mixture of carbon fiber and epoxy is reached, the excess solvent is then distilled for recycled use. By redistributing the orientation of the fibers within the epoxy matrix this would yield a high strength and flexible material that could be used in the form of a sheet molded compound. The fibers are placed into a hot press for cured bulk molding that can then be characterized through optical microscopy, thermal gravimetric analysis (TGA) and/or mechanical testing. The data collected through these methods is compared to the industry standards for sheet molded compounds made from unaged carbon fiber material. Completion of this research would result in a novel bulk molded compound through a fiber reutilization process. 

Endophytes, microbes that colonize inside plants, may promote plant growth by several mechanisms, such as providing nutrients, defending against pathogenic microbes, and producing phytohormones, that help the host plants tolerate environmental stresses. Using endophytes to promote plant growth could reduce agricultural fertilizer use, a major source of nutrient pollution due to runoff. Endophytes that inhibit plant pathogens may also reduce germicide and fungicide use, which can be toxic to non-target organisms. We used bioinformatic tools to analyze the genomes of the endophytes Azospirillum sp. 11R-A, Herbiconiux sp. 11R-B1, Rhizobium sp. PTD1, Rahnella aceris R10, Azotobacter beijerinckii SD1, R. aceris WP5, and Sphingobium sp. WW5 for the genetic features associated with nitrogen fixation, phosphate solubilization, production of the phytohormone indole-3-acetic acid (IAA), and the synthesis of antimicrobial secondary metabolites, and then used the genotype characterizations to predict whether those traits would be observed in their phenotype.

Based on their genotypes, strains 11R-A, R10, SD1, and WP5 were predicted to fix nitrogen, however, only SD1 grew on nitrogen-free agar. 11R-A, PTD1, R10, SD1, and WP5 were predicted to solubilize phosphates, but only R10 and WP5 solubilized it. 11R-A, 11R-B1, and PTD1 were predicted to produce IAA, but 11R-A, 11RB-1, and WW5 did so in vitro. All strains except for WP5 were predicted to make antimicrobial secondary metabolites.

We identified several discrepancies between the genotypes and phenotypes of nitrogen fixation, phosphate solubilization, and IAA production. In some cases, the phenotype was not present despite the genotype indicating it. This may be because the genes weren’t under the right conditions to be expressed in vitro. A potential next step would be to determine the conditions under which these genes are expressed to predict if the strains are good candidates for plant inoculation. In other cases, the phenotype was present even though the genotype didn’t indicate that it would be. This may be an issue with the genome annotation software missing certain genes or the presence of an unknown pathway.

Inoculating plants with optimized plant growth-promoting endophytes may improve host-plant growth in harsh environments while minimizing chemical and energy inputs.

Engineered microbes provide an environmentally friendly path to chemical industries, including fine chemicals and therapeutics. To effectively modulate such microbes, genetic circuits can be utilized as a programmable tool to control multiple genes within microorganisms. CRISPR-mediated gene activation (CRISPRa) is an emerging tool suitable for this purpose. In CRISPRa, a nuclease-deficient dCas9 protein is used to recruit a transcriptional activator domain (MCP-SoxS) to activate  genes of interest. Using a complementary guide RNA (gRNA), CRISPR-dCas9 complex can be directed to nearly any DNA target. Despite the programmability of CRISPRa, the maximum number of genes that can be simultaneously regulated remain unexplored. In this work, we aim to experimentally investigate the limitation on the number of gRNAs in the chemical bioproduction context. First, we designed CRISPRa circuits consisting of 1 to 6 gRNAs encoded on plasmids constructed with a scalable and high-throughput technique based on Golden Gate Assembly. CRISPRa circuit performance was then evaluated by simultaneously regulating multiple fluorescent proteins as a proxy for multi-enzyme cascade in biosynthetic pathways. Increasing the number of gRNAs was found to decrease CRISPRa activity, suggesting competition of CRISPRa components. Furthermore, we applied the constructed circuits for metabolically engineered pathways in Pseudomonas putida regulating production of p-aminocinnamic acid (pACA), a precursor for polymer synthesis vital in photovoltaic and biomedical applications. Bioproduction of pACA in P. putida was enabled by simultaneously regulating 6 heterologous genes. The outcome of CRISPRa circuits will be analyzed via High-Performance Liquid Chromatography (HPLC). This work demonstrates the constructions and implementations of multi-gRNA CRISPR circuits in bacteria, and expands the capabilities of CRISPR circuits in a wide range of biosynthetic applications. 

Throughout the world, cannabis is the most universally used psychoactive drug. The National Survey on Drug Use and Health (NSDUH) reported in 2020 that more than 48 million Americans ages 12 and older used cannabis in the past year. Since cannabis use is most popular among people between ages 18 and 35, an understanding of the consequences cannabis use on reproductive functions in offspring is crucial since the majority of consumers use cannabis during their most active reproductive ages. Additionally, an increasing number of pregnant mothers are using cannabis. It is undetermined whether the negative effects of cannabis use on reproduction are passed to subsequent generations due to germline transmission. We propose that in utero and nursing exposure to cannabis vapor effects the reproductive systems of offspring. The impact of cannabis vapor exposure during pregnancy and after birth was assessed through analysis of the gonads, steroid hormones, and sperm count and motility of the next offspring (F1). CD-1 female mice were mated with drug naïve male mice. Pregnant females (F0) on gestational day 1 (GD1) were exposed to control vehicle (80% propylene glycol and 20% grape seed oil), 100 mg/ml, or 200 mg/ml of cannabis extract for 30 minutes twice a day. Sperm count and motility, testes weight and histology, and body weight were assessed. Our data indicates that sperm count and motility was significantly reduced following 100 mg/mL and 200 mg/mL cannabis vapor exposure, when compared to vehicle.  No significant difference in sperm count or motility was observed between 100 mg/mL and 200 mg/mL treatments. Both testes weight and body weight were normal. This research is a starting point in examining the generational impact of cannabis vapor exposure on male reproductive systems. In summary, the results suggest in utero and nursing cannabis exposure affect male reproductive functions, leading to subfertility.

In construction materials, cementitious grout is used as a filler or a sealant, whereas cement is its major mixing ingredient. Cement produces high CO2 emissions and is a major environmental problem. Therefore, introducing admixtures in the grout decreases CO2 emissions and improves its properties. One known grout admixture is cellulose ether, but it has a high market cost. Therefore, biochar is introduced as a potential partial replacement for cellulose ether due to its cheap process. Thus, this study investigates the properties of cementitious grout of cellulose ether, which has been partially or completely substituted by biochar. Four formulations were prepared by maintaining cement, silica sand, polymer powder, and distilled water at a constant. The formulation for “Sample 1,” the control sample, only contained cellulose ether. The formulation for “Sample 2” only had biochar, the formulation for

Sample 3” contained 50% of cellulose ether and 50% mixed biochar produced at 300 and 500 °C, and the formulation for “Sample 4” had 50% cellulose ether and 50% of acidic biochar. Each formulation sample was subjected to different tests that analyzed its surface chemical composition (via FTIR), workability, compressive strength, elemental composition, morphological structure, thermal stability, and cement hydration. The results showed that “Sample 3” increases cement hydration, workability, and compressive strength by up to 85% compared to the control formulation. Therefore, this study concludes that the appropriate combination of biochar and cellulose ether improves the properties of cementitious grout.

Prenatal exposure to infection and excessive stress is a contributing risk factor in developing several psychological disorders, including Schizophrenia, depression, and alcoholism. In addition, it leads the offspring to have a history of alcohol misuse due to viral stress being introduced during prenatal development. Little research has been done to understand how prenatal exposure to infection impacts brain development leading to the propensity to develop alcohol use disorder. Our lab has demonstrated the utility of the antioxidant treatment of N-Acetylcysteine (NAC) in the reduction of alcohol seeking behavior. We hypothesize that NAC can prevent adult alcohol use behavior. Using pregnant Sprague-Dawley rats, we applied maternal immune activation (MIA), a model of prenatal exposure to infection combined with adolescent alcohol exposure (AE) which leads to increased drinking behavior in adulthood. Pregnant Sprague Dawley rats were exposed to NAC or saline 24 hours before and after the MIA treatment. After birth, adolescents were given 24-hour home cage access to 10% ethanol (AE) using a two-bottle choice technique. In adulthood, rats were trained to press an active lever to self-administer 10% ethanol for 30 minutes, 5 days per week. Data from our lab indicate that prenatal exposure to stress impacts alcohol self-administration in rodents. We show that NAC treatment reduced self-administration in MIA females, and in MIA males; there was a slight increase in preference for active/inactive lever presses in self-administration. These data suggest that there is a sex link to adolescent drinking behavior following maternal immune activation in the womb leading to increased adult alcohol seeking behaviors in males. NAC exposure decreases the effect of MIA and reduces alcohol seeking behaviors in offspring. Overall, these data demonstrate the utility of NAC in decreasing alcohol intake in this model and the mechanism by which MIA alters drinking in adult offspring. 

Obesity is associated with numerous health risks, including type 2 diabetes, cancer, arthritis, high blood pressure. A major contributor to the obesity epidemic is the ease of availability and overconsumption of foods high in fat. Our lab and others have shown that exposure to foods high in fat causes adaptations in the prefrontal cortex (PFC), a region of the brain critical to executive functions such as decision making. The food-induced adaptations within the PFC following exposure to palatable diets may play a key role in driving maladaptive food seeking behaviors. Our lab has recently shown that exposure to foods high in fat alters perineuronal nets (PNNs) within the PFC of Sprague Dawley rats. Within the PFC, PNNs primarily surround GABAergic fast-spiking interneurons responsible for maintaining the excitatory/inhibitory balance of the local neuronal circuit. PNNs are specialized extracellular matrix structures that contribute to synaptic stabilization as development progresses and are comprised of chondroitin sulfate proteoglycans (CSPGs) such as brevican, neurocan, versican, and aggrecan. While brevican. Neurocan, and versican were found to be derived from glial cells, aggrecan is primarily derived from neurons. Our hypothesis is that the CSPG components, brevican, neurocan, and versican will be attenuated following exposure to a diet high in fat due to their dependence on glial cells, which play a critical role in inflammation. To test our hypothesis, we are using multiple techniques including Western blotting, mass spectrometry, and immunohistochemistry to identify the components of PNNs that may be impacted by high fat exposure. To date, we have qualitatively identified protein bands reflective of molecular weights corresponding to the CSPGs discussed above and are currently having those bands verified with mass spectrometry. Our ultimate aim is to selectively target these CSPGs to modify maladaptive food seeking behaviors that may contribute to the development of obesity.

Within sarcomeres, the most basic unit of muscles, thin and thick filaments slide against one another to contract the muscle. Actin and tropomyosin compose the main structure of the thin filament while two homologous proteins, leiomodin and tropomodulin, compete for binding sites at the pointed end of the filament. Leiomodin forms a leaky cap that allows for the elongation of thin filament proteins while tropomodulin prevents the lengthening of the filament. This modulation of the filament is vital to the function of the muscle as any significant deviation from uniformity within cardiac muscles can result in cardiomyopathy. Within our research, our objective is to understand binding behaviors between leiomodin and tropomyosin in order to better form a model of thin filament length regulation. Specifically, we hypothesized that the regulation of thin filament length is based primarily on the differences in affinities of the two homologous proteins for tropomyosin. To test this, we engineered and introduced point mutations into a peptide fragment of the cardiac leiomodin isoform representing its tropomyosin binding site (Lmod2s1) and analyzed structural changes in complex with tropomyosin through circular dichroism and molecular dynamics simulations. We found that Lmod2s1 peptide fragments with point mutations such as Lmod2s1 S24K, and E36V provided differing affinities that, in conjunction with prior studies and future studies into length of filaments versus affinity of leiomodin, will allow us to test if our hypothesis is indeed true.

Our goal is to provide technological support for automated cognitive health assessment and intervention. One type of self-report assessment is a person's response to "in the moment" ecological momentary assessment (EMA) questions. We hypothesize behavior context can be used by a machine learning algorithm to predict responses to EMA questions.

In this project, we extract digital behavior markers from continuously-collected Apple Watch data for persons with cognitive limitations and their caregivers. We evaluate the ability of the learning algorithms to predict EMA responses from the behavior markers and assess the types of behavior contexts most indicative of stressful and non-stressful interactions. The machine learning algorithm will be captured using smartwatches and machine learning, using other related methodologies of ambient sensors to capture data. The project is at the very early stages in our timeline; we are heading in different directions and will soon narrow down our outline to ensure data collection will be most affected and be able to give a strong interpretation of what we find.

Thornton Creek has a drainage area of 12 square miles in northeast Seattle. Due to its location, it is susceptible to pollution from water runoff that comes from surrounding roads. Historically, Seattle has used various road de-icing compounds, such as calcium chloride in a soy-base, sand, and eventually sodium chloride after a 2008 snowstorm that shut down the city. During the 1990s, various habitat restoration projects took place, and the surveys taken to measure project efficacy discovered that Coho salmon were dying before they could make it back to their natal streams to spawn. Researchers studied this occurrence and ultimately attributed salmon mortality to toxic runoff from urban land usage. This study aims to investigate the effects of using sodium chloride to de-ice roads on salmon livelihood. Water samples were collected two to three times daily for a week from Thornton Creek during Seattle’s ice storm in December 2022. Then, sodium chloride concentrations were tested using a Metrohm 930 Compact Flex IC in February 2023. The results from these tests were compared to the EPA’s sodium chloride toxicity levels, and none of the samples exceeded the chronically contaminated sodium chloride concentrations for a single event. Although the results from these samples do not indicate chronic contamination, further research is necessary to determine the long-term effects of using sodium chloride to de-ice roads. Through continuous monitoring of sodium chloride concentrations, the findings of this study can aid in finding alternatives to road salt that have less of an environmental impact to maintain water quality and improve the livelihood of salmon that spawn locally in Seattle.​

Introduction

Pediatric chronic pain is increasingly prevalent. While multidisciplinary pain treatment (MPT) has shown success in intervening, MPT often requires frequent attendance that can be impacted by disparities in care access. Improved understanding of factors that predict service access and associated health outcomes can inform screening and intervention efforts.

Methods

Youth participants (N=424) had 1) a chronic pain diagnosis and 2) engaged in a pediatric pain evaluation (PE) by a multidisciplinary pain team [psychology, anesthesiology, physical therapy (PT)]. Retrospective electronic health record (EHR) collection captured patient demographics, parent-reported social risks, youth post-PE service access, and acute medical visits (ED visits and hospital admissions) one year prior to and post PE.

Results

Youth experienced significantly fewer ED visits and admissions in the year following PE compared to the year prior (all reported findings p<.05), although youth with above-average life stress were more likely to experience ED visits post PE. Following PE, approximately 60% of youth were able to access recommended PT and pain psychology services. Regarding social factors, youth with married caregivers were significantly less likely to have ED visits prior to and post PE and youth of color (identified by EHR with a Hispanic ethnicity and or non-white race) experienced more acute events prior to and post PE.

Conclusions

MPT access was associated with fewer acute visits, with specific social factors predicting health outcomes. The higher frequency of acute medical events experienced by youth of color prior to, and post PE access represents a disparity potentially driven by discrimination in referral processes, care provision and other system factors – resulting in unequal access and health. These findings highlight the importance of addressing disparities in pediatric chronic pain care access, considering social risk in screening practices, and tailoring interventions to account for such risk.

Eukaryotic cells (human/animal cells), undergo a process of water exchange across the cell membrane to carry solutes into and out of the cell and to remain in a state of homeostasis. The phenomenon of water exchange is made up of 2 components, a passive component in which water simply diffuses across the membrane without the need of energy, and an active component in which ATP is required to cycle water into and out of the cell. The AWC hypothesis states that the rate of water exchange is driven by the concentration of ATP. Water exchange can be measured using magnetic resonance, specifically NMR for in vitro experiments and MRI for in vivo experiments. It has been found that active water exchange decreases in cancer cells, and in yeast cells undergoing the process of fermentation. Our lab uses yeast as a model organism to elucidate the mechanism by which metabolism influences water exchange. Respiratory modulators have been employed to directly test the impact of mitochondrial activity on water exchange.

High concentrations of the respiratory modulators may be harmful to cells and require organic solvents as a drug delivery vehicle. The focus of my research is to measure cell viability in response to the presence of respiratory modulators and organic solvents as an experimental control, and to quantitatively determine the correlation between water exchange and ATP concentration of the cell. Our group measures the ATP concentration using a luciferase assay where the amount of light detected from our sample is used to calculate the amount of ATP present.

Rationale: P2X3 purinoceptors are expressed by airway sensory neurons and are activated by extracellular ATP released during periods of cell stress. In asthma, sensitivity to ATP is increased. Airway eosinophilia, which is common in a majority of asthmatics, increases airway epithelial sensory nerve density in mice and in humans. Whether eosinophils increase neuronal P2X3 expression in asthma is unknown.

Methods: P2X3 expression was quantified in sensory neurons in vagal ganglia from wild-type (WT) mice, transgenic mice with chronic pulmonary eosinophilia (NJ1726 lineage), transgenic mice with chronic systemic eosinophilia (NJ1638 lineage), and eosinophil-deficient mice (PHIL) using RNAscope in situ hybridization. Images were obtained using an ApoTome confocal microscope (40X, 1.3 N.A.). P2X3 expression was determined by measuring the percentage of P2X3 positive pixels within three, randomly assigned, non-overlapping 50x50 micron sections of vagal ganglia using ImageJ. Replicates were averaged to generate a single data point for each animal.

Results: P2X3 RNA was detected in vagal ganglia tissue sections in all mouse groups. P2X3 RNA expression was present in 52.1 ± 4.0% of vagal neuron tissue analyzed in WT mice, 72.9 ± 6.1% in mice with chronic pulmonary eosinophilia, 66.7 ± 5.5% in mice with chronic systemic eosinophilia, and 59.3 ± 4.7% in eosinophil-deficient mice.

Conclusions: P2X3 is highly expressed in vagal sensory neurons. Increased P2X3 expression in the setting of chronic eosinophilia suggests eosinophils positively regulate sensory nerve purinoceptor expression. Eosinophil effects may contribute to increased ATP sensitivity in humans with eosinophilic asthma.