Many human diseases, including cancer metastasis, and developmental disorders, involve sheets of cells moving together. This concerted process, called collective cell migration (CCM), is critical to maintaining the integrity of the sheet of cells during movement. Maintaining cell sheet integrity is dependent on individual cells retaining their shape while still connected to neighboring cells. To study collective cell migration, we use Drosophila melanogaster (fruit fly) embryogenesis. Dorsal closure (DC), an event in Drosophila embryogenesis, is an example of CCM. During DC, two epithelial layers wrap around the embryo. Because CCM’s core mechanics are conserved, studying DC provides an opportunity to indirectly study human diseases/disorders function in a model that has fewer genetic redundancies and is easy to manipulate. Adherens junctions (AJs), are the site of adhesion between individual cells, and what allows for dynamic rearrangements like CCM by strategically increasing or decreasing adhesion. Despite the importance of this process and its relevance to many diseases, CCM is highly complex and how the scaffolding protein Canoe stabilizes adherens junctions under high stress is not entirely understood. We are interested in how junctions between cells are stabilized during a dynamic movement like in CCM. Using a maternally driven Gal4/UAS system, we created moderate canoe knockdowns. We then performed targeted, immunofluorescence antibody staining of embryos during DC. We expect that the loss of Canoe will lead to increases in cell-sheet tension as well as the compensation by other stabilizing proteins like Polychaetoid. Our research improves our understanding of stabilizing proteins at AJs and CCM and may lead to the more effective treatment of disrupted CCM human diseases/disorders like wound healing, palate formation, and neural tube closure.

I am furthering research in growing successful plant life in Martian regolith. The intention of this research is to amend Martian regolith so that it can support edible plant growth. We intend to counteract the characteristics that would impede plant growth and introduce low-mass additives that support plant growth. To simulate Martian ground “MMS-2” is used, this regolith is designed to be over 90% similar in chemical and textural composition to ground cover on Mars.Challenges noted from previous research include inhibited root growth, inefficient nutrient absorption, and impeded water uptake due to clay-like consistency. The additives selected to counteract these challenges include vermiculite to expand the soil and improve water circulation, and Biodyne™ microbes to enhance nutrient uptake. I chose to also incorporate mycorrhizal fungi to support and enhance root development and function. My previous research indicated that mycorrhizal fungi played a significant root development, plant height, and leaf growth. My current research is focusing on reproducing these results with more specimens and clear data points to further evaluate the impact of these additives. Rosemary was selected due to its ability to thrive in harsh, cold, and dry climates. As this plant is notoriously difficult to grow from seed, seeds were pre-sprouted to confirm viability prior to planting in the experimental pots in order to assure more specimens. Experimental regolith are treated with mycorrhizal fungi, Biodyne™, or both receive treatment mixed in water; all regolith and the control are based in a 30/70 earth soil to regolith blend with vermiculite. Plants treated with mycorrhizal fungi are anticipated to have darker and plentiful leaves; plants treated with Biodyne™ are anticipated to show straighter growth, plants treated with both are anticipated to be the most successful with both plentiful dark thick leaves and straight strong growth.

We are expanding previous research which manipulated the composition and chemistry of Mars regolith to better support plant life. The Mars regolith we are using is a 70% regolith 30% compost mix. Our focal plant for this experiment is Salvia rosmarinus (hereinafter rosemary) because of its resilience in cold climates and minimal water needs. Rosemary has been observed to have low germination success in both Earth soil and Mars regolith, to address this we sprouted seeds in water prior to transplant in order to guarantee seeds were viable rather than sowing rosemary directly in Mars regolith. We will ensure cotyledons have developed and stems have reached <5.0mm in length prior to transplanting in regolith to eliminate the variable of low seed viability. We hypothesize the amendments made to Mars regolith along with microbial and mycorrhizal additives will significantly improve rosemary development and growth. Plants we have previously attempted to grow in Mars regolith experienced difficulty establishing roots when transplanting cuttings and when grown from seed due to the clay-like consistency of the regolith. We have adjusted the previous experimental design by adding vermiculite to improve drainage and reduce regolith density, microbes to fixate Nitrogen and assist in nutrient uptake, and mycorrhizal fungi to encourage more effective root establishment and development. Previous experiments with Biodyne™ microbes and mycorrhizal fungi in Mars regolith have shown promising results in kale and rosemary. We anticipate the addition of Biodyne™ and mycorrhizal fungi, both independently and together, will improve root development, plant height, leaf growth, and chlorophyll richness. To test this, we will collect data bi-weekly by noting plant “greenness” and measuring plant height, leaf length, leaf count, and root growth.

Kaposi’s Sarcoma (KS) is a highly vascularized tumor, which affects AIDS patients worldwide and remains endemic to sub-Saharan Africa. Kaposi’s Sarcoma-associated Herpesvirus (KSHV) is the etiological agent of KS and its latent infection is involved in tumor formation and the induction of angiogenesis in the spindle cell, a cell of endothelial origin and the main proliferating cell type in a KS tumor. Previous RNA-Seq data obtained by our group showed that osteopontin (opn), a secreted protein known to act as a ligand for integrin receptors that activate signaling cascades that promote angiogenesis, is highly upregulated at the transcript level during KSHV latent infection of endothelial cells. To determine whether opn is required for the activation of angiogenesis in KSHV latently-infected endothelial cells, we used CRISPR-lentiviral constructs to knock out opn and evaluate changes in angiogenic phenotypes upon KSHV infection via cell proliferation, tubule formation, and cell migration assays. Preliminary results reveal a significant reduction in tubule formation in opn knockout KSHV-infected endothelial cells. This finding suggests that opn upregulation in such cells is responsible for the activation of this angiogenic phenotype. Future experimentation will include evaluating how KSHV induces the upregulation of opn by infecting wild-type endothelial cells with mutant viruses lacking certain latency protein genes and evaluating differences in opn transcriptional and translation, as well as evaluating the mechanisms by which opn activates tubule formation in KSHV latently-infected endothelial cells. Identifying  the drivers of angiogenesis in KSHV-infected endothelial cells will aid the characterization of therapeutic targets for KS progression in such cells, given that KS tumors are highly angiogenic from its early stages.

Microbiomes are now known to provide numerous benefits to hosts, from immunity to reproductive success. Vertical transmission of microbes has been linked to improved offspring success, for example by protecting eggs from pathogens post-oviposition. Infection with pathogenic soil fungi is associated with mortality in soil-incubated eggs, but may be ameliorated by the transmission of beneficial microbes.To determine whether 1) maternal microbes are deposited on eggs during oviposition, and 2) whether they have an antifungal effect that increases hatching success, we conducted challenges between dissected or oviposited eggs from the striped plateau lizard (Sceloporus virgatus) and fungi from failed S. virgatus eggs. Unique fungal and bacterial morphs were isolated from eggs to identify potential pathogenic fungi and antifungal bacteria. Direct challenges were carried out between individual bacteria and fungi to assess antifungal capabilities of bacteria isolated from the S. virgatus eggshell microbiome. Dissected eggs (no contact with the cloaca) had significantly lower hatch success, and hatchlings from dissected eggs were significantly smaller and lighter than those from oviposited eggs, indicating that some element of oviposition is key for hatchling success. One bacterial genus (Enterobacter) was found in the cloacae of females and on eggshells, suggesting that vertical transmission may be occurring. Three fungi were found to dominate the eggshell community of both dissected and oviposited eggs exposed to multiple fungi, suggesting that they may be responsible for egg mortality. At least two of these species of fungi have known pathogenic effects in other reptiles, and may produce antibacterial and antifungal metabolites.These results suggest an antagonistic relationship between fungi and bacteria on S. virgatus eggshells, indicating that bacteria transmitted during oviposition may improve hatch success in this species. Additionally, vertical transmission of beneficial microbes may serve as a form of parental care and occur via a novel pathway, the cloaca.

Many widespread species show morphological variation across the unique habitats and climates they inhabit. For reptiles these morphological adaptations such as limb length or body depth are often correlated with habitat use and substrate types. Additionally, body size trends and the degree of pigmentation of reptiles, driven by thermoregulatory requirements, can correspond with the elevation and climate of populations. The blue ventral coloration of Western Fence Lizards (Sceloporus occidentalis) is a social signaling device and can also be a byproduct of increased melanism in response to thermally challenging environments. I used handheld and photo analysis methods to obtain head, body, and limb measurements in addition to quantifying belly and throat coloration from S. occidentalis in Washington State at a high elevation creek canyon population, a low elevation forest population, and a coastal beach population. I found that high elevation creek canyon lizards used rock perches 69% of the time, low elevation forest lizards used trees 67% of the time, and beach lizards perched on driftwood 97% of the time. Given this distinct substrate use, my morphological data suggests adaptations between populations. Rock lizards had the longest front fore-limbs which could result in increased running speed to optimally navigate their open rocky habitat. Forest lizards had the shortest front fore-limbs possibly due to reduced running reliance, as they specialized in arboreal perches. Driftwood lizards had the skinniest body condition that could optimize them for cracks in driftwood logs they relied on. Beach lizards had bluer and darker bellies than high elevation lizards, and bluer throat patches than the other two sites. My study suggests morphological and signalling adaptations among populations of S. occidentalis at the species’ geographic extreme, and demonstrates how habitat and climate selection pressures create intraspecific variation between populations of widespread species.

Thirteen months after the first confirmed case of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the United States (U.S.), over 500,000 people have died. However, the pandemic in the U.S. has not affected all populations equally; there are vast differences in morbidity and mortality in areas of differing population densities. We hypothesized that the number of SARS-CoV-2 mutations would be higher in densely populated areas due to closer proximity among inhabitants, which would lead to increased viral spread from person to person, and thus a greater number of mutations. These mutations can impact and reduce vaccine efficacy, as well as morbidity and mortality, depending on where they occur in the virus’s genome. The complete genomes of SARS-CoV-2 cases from March 1^st, 2020 to March 1^st, 2021 were collected from the Global Initiative on Sharing Avian Influenza Data (GISAID) for counties of varying population density. These genomes were analyzed to identify geographic areas where problematic mutations had the potential to occur. Demographic data was collected at the county level from Integrated Public Use Microdata Series (IPUMS) for additional analysis. Mapping the incidence of mutations in the SARS-CoV-2 genome and the correlation of these mutations with population density and other demographic indicators may help decrease disease spread and ensure the vaccines will remain effective.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic emerged in the United States in January 2020, altering how most individuals interact in public spaces. Many familiar indoor spaces such as restaurants, planes, and classrooms suddenly posed a significant risk of exposure to SARS-CoV-2. Most transmission of SARS-CoV-2 is airborne through contagious aerosols exhaled with carbon dioxide (CO₂) by infected individuals in indoor and outdoor spaces. Indoor CO₂ levels are impacted by factors including: size of the space, air changes per hour, number of individuals present, the activities of the individuals, humidity, and temperature. Vigorous activities — such as speaking or exercising — increase CO₂ levels just as they increase aerosol production. Thus, CO₂ levels provided a good approximation of the transmission risk of SARS-CoV-2 in a specific location. We investigated how available ventilation impacts the transmission risk of SARS-CoV-2 in different indoor settings. An indoor air quality monitoring system that utilized CO₂ levels was developed to evaluate risk of transmission. CO₂ levels were measured using CO₂ sensors in Seattle, WA from March 2021 to April 2021. This data was collected in a variety of public indoor spaces including public transit, educational buildings, restaurants, gyms, and grocery stores. CO₂ levels were analyzed with a COVID-19 Aerosol Transmission Estimator based on the Wells-Riley equation. This information is critical to understanding the comparative risk of contracting SARS-CoV-2 in various indoor public spaces and highlighting where improvements can be made to mitigate such risk.

Every Spring, a small group of gray whales, nicknamed the “Sounders,” come to Possession Sound and the surrounding waters to feed. Many of these individuals have been documented in this region consistently since the 1990s. Although this group of less than twenty individuals has been the subject of several research studies, little is known about their movement patterns in Possession Sound. Over the past few years, gray whales along the West Coast have been experiencing a mass mortality event. Gray whale research is essential in identifying potential contributing factors. The Sounders are benthic feeders and primarily consume ghost shrimp that live in the sediment throughout the intertidal zone. These gray whales access a handful of shallow areas for feeding which are only accessible during high tides, such as the Snohomish River delta. This research evaluates and combines fifteen years of sighting data from Ocean Research College Academy (ORCA) and thirty years of sighting data from Cascadia Research Collective (CRC) with the intent of identifying spatial patterns and possible correlations with the tides. I am creating ORCA’s sighting catalog with photographs taken during ORCA’s sightings and identifying the individuals present using reference photographs from CRC and compiling any additional available data. In this catalog, sighting data is evaluated on individual whale identification, date, time, location, and additional observational notes such as whale interactions. I am plotting this data and additional data from CRC’s sighting catalog in ArcGIS to create a map of spatial distributions. Preliminary results from ORCA’s sightings suggest that gray whales are most often sighted in Possession Sound between Hat Island and the south end of Jetty Island. Any spatial trends that I identify may lead to a better understanding of gray whale behavior, possibly including gray whale intercommunication, social structure, individual behavior, and local feeding practices.