With this project, researchers for the Harry Bridges Center for Labor Studies from Seattle, Tacoma, and Bothell campuses seek to gain a better understanding of the intricate relationship that exists between the UW undergraduate experience and employment, as well as inform students on existing resources and support services both on and off campus. To sufficiently measure unique student experiences, we have sought survey responses through canvassing, contacting departmental and Registered Student Organization (RSO) listervs, and visiting in-person classes. Over the course of the past 6 weeks, we have made 73 points of outreach at the Seattle campus, 62 at Bothell, and 44 at Tacoma, resulting in 430 survey responses as of February 7th, 2024. Our Qualtrics survey will run through the end of winter quarter. We will then also offer paid interviews for 15-20 students to supplement survey data through further examination of survey themes and trends. Modeling UCLA Labor Center’s Unseen Costs: The Experiences of Workers and Learners in Los Angeles County, we will then compile our survey results and interview stories into a cumulative research report about why UW students work while pursuing their undergraduate degrees, what fields they have worked in, and any associated workers’ rights violations they may have faced while in these positions. With this research, we aim to create a more supportive environment for our student workers, and to help educate our respective campuses to meet their student body needs.

During neonatal transport, specialized pediatric transport teams closely monitor the status of critically ill newborns. Hyperspectral imaging, a method of manipulating light, can be used to measure the vital signs and video the patient’s physical appearance for remote monitoring. Appropriate light intensity is critical for clear visibility of the newborn and hyperspectral imaging accuracy, but this must be balanced with safety for sensitive eyes. My previous studies have determined the minimum range of light needed to accurately view the neonate in a transport incubator. In addition, I conducted a literature review to understand the uses of hyperspectral imaging in dermatology and surgical fields and to gain knowledge of the imaging equipment and biological signal processing. The experiment was designed to use hyperspectral imaging to gather vital sign data using a near-infrared camera with the purpose of using hyperspectral imaging in a dimly lit environment like that experienced on transport. In this experimental design, physiologic data of breathing rate and blood oxygen levels were recorded using 1-minute-long videos, while blood oxygen and heart rate was collected using a pulse oximeter placed on the fingertip of the subject. Following the data collection, the experiment is designed to extract the biological signals through filtering of two different ranges, a beats per minute range for the heart rate and a blood oxygen level range for the blood oxygen levels. Data analysis aims to compare the vital sign data collected using hyperspectral imaging and that using a pulse oximeter to understand the feasibility of the hyperspectral imaging for vital sign extraction. The expected result of this study is that the heart rate and blood oxygen levels of the patient measured using light and a pulse oximeter will be similar. In conclusion, this research will demonstrate the potential application of hyperspectral imaging to neonatal transport.

When a cell undergoes stress conditions, such as oxidation or aging, an increase in protein instability can occur and prevent proper cell functions. Small Heat Shock Proteins (sHSPs) are molecular chaperones that work to maintain a healthy proteome by associating with misfolded “client” proteins to delay aggregation under such conditions. HSPB5, a human sHSP, is ubiquitously expressed throughout the body. HSPB5’s disease mutant, R120G, is a defective chaperone associated with cataracts and desmin-related myopathy. It is still unknown how this mutation is detrimental despite many years of research. My research aims to understand how this mutation retunes the electrostatic properties of HSPB5, affecting its chaperone activity. Residue R120 is part of an electrostatic network that helps create an important structural feature in the folded region of HSPB5, the alpha-crystallin domain (ACD). In the unmutated (WT) protein, the ACD surface is overall positively charged. Substitution of the positive R120 to glycine alters both ACD’s structure and electrostatics. I generated two mutants, R120K (retaining positive charge) and R120D (switching to negative charge) to investigate how R120 plays a role in ACD’s conformation. Using a negatively-charged molecule, ATP, as an “electrostatic” probe in 2D NMR, I observed differences between its binding affinity to my R120 variants. I found that only R120K ACD behaves similar to WT ACD, suggesting a possible correlation between charge potential and ACD’s interactions with ATP. Currently, I am investigating if charge potential affects chaperone activity through aggregation assays with a client protein, human γD-crystallin, found in the lens and implicated in cataracts. I predict that WT and R120K, with similar electrostatic properties, will have similar chaperone activity. R120G and R120D, prevalently in an “active” state, will have higher chaperone activity. Understanding how such mutations affect HSPB5’s conformations and chaperone activity is a step forward in understanding sHSPs’ chaperone mechanism.

Small heat shock proteins (sHSP) are a family of molecular chaperones whose function is to delay the harmful aggregation of other proteins. Protein aggregation is associated with neurological disorders such as Alzheimer's disease and Parkinson's disease. In many tissues, multiple sHSPs are coexpressed and tend to assemble into hetero-oligomers. Hetero-oligomers are complexes of two or more different protein species. The extent and mechanism by which these hetero-oligomeric complexes form is yet to be fully understood. The goal of my discovery-driven research is to assess how the properties of sHSP hetero-oligomers differ from the properties of homo-oligomers. In my project, I focus on three sHSPs that are highly expressed in muscle: HSPB1, HSPB5, and HSPB6. Each of these proteins exhibit different behavior when on their own. HSPB1 and HSPB5 form a distribution of large homo-oligomers, whereas HSPB6 forms a small homo-dimer. One of the most characteristic properties of the small heat shock proteins is formation of oligomers that span different sizes. Thus I am primarily determining the sizes and composition of the sHSP hetero-oligomers. I performed a comprehensive study to characterize the sizes of the hetero-oligomers using three complementary methods: analytical size exclusion chromatography, mass photometry, and native gel electrophoresis. I have found that HSPB6 is able to readily incorporate into hetero-oligomers as the concentration of the other sHSP is increased, and that the complexes are formed in a distribution of intermediate sizes. I am currently working on assessing the ability of the hetero-oligomers to act as molecular chaperones by aggregation assays. I predict the hetero-oligomers will delay protein aggregation more efficiently than HSPB6 on its own. The findings of my project give insight into why sHSPs are coexpressed and form hetero-oligomers in cells. Understanding these hetero-oligomers sheds light into the complex pathways of sHSP function. 

Despite the extensive literature on Canada and European countries, there is relatively little research on paid maternity policies in U.S. states due to its recency. Economic theory would suggest that fertility rates would increase with a paid maternity leave policy as the cost of having a child would be effectively subsidized. A strong family leave plan could be the deciding factor for many families to have a child. However, current research is inconclusive with there being no studies on U.S. fertility rates specifically in mind. The goal of my research is to provide insight on the way in which the U.S. fertility rate may be affected by paid leave programs. To accomplish these goals, I will be studying the implementation of paid maternity leave in Washington state specifically. I will be using a difference in difference method to measure the effect of this policy. I will be using data from the National Center for Health Statistics and the American Community survey for my fertility rate estimates. My research is in progress; however, I would expect either slight or no change in fertility rates in Washington based on previous research in the field. The goal of this research is further insight to nationwide implementation of this program by analyzing a specific possible implication of this policy and to contribute to the literature of paid maternity leave on fertility rates.

Diffusible iodine-based contrast-enhanced micro-CT (diceCT) is a technique used to image soft tissue specimens using 3D x-ray microscopy. Staining soft tissues with iodine (I2KI) solution prior to scanning improves contrast for detailed visualization of internal organs, but iodine staining is also associated with tissue shrinkage which can interfere with quantitative analysis. It has been shown that stabilizing soft tissue with hydrogel can reduce shrinkage. We adopted these protocols for our lab, but, despite using hydrogel stabilization, we observed wrinkles in the external surfaces of E15.5 mouse embryos, qualitative evidence of considerable shrinkage. To quantitatively test for shrinkage, we compared the crown rump lengths (CRL) of mouse embryos measured from photos taken prior to the scanning process and then from diceCT scans. CRLs ranged from 12.4 to 20.0 mm in photos and 11.1 to 16.8 mm in scans. An average reduction of 12% resulted from the specimen preparation process and confirmed tissue shrinkage. Furthermore, the amount of shrinkage was not uniform across the specimens, complicating quantitative analysis based on diceCT. Our first hypothesis was that the iodine solution used to prepare the specimens was too acidic. We measured the pH of this solution and found a range from 4.5 - 6.4. To examine if a neutral pH reduced tissue shrinkage, we prepared specimens with a buffered iodine solution (pH 7.2). DiceCT scans of embryos in buffered iodine solution did not show reduced shrinkage compared to controls in the original solution. Further investigations will focus on other potential sources of shrinkage including the pH of other solutions and the time specimens spend in each step of the protocol. Continuing to investigate sources of tissue shrinkage in diceCT can lead to additional methods for shrinkage reduction, supporting more accurate quantitative analysis of diceCT.

Electron paramagnetic resonance (EPR) spectroscopy is a powerful and versatile spectroscopic tool for detecting unpaired electrons in molecular systems. We are particularly interested in using EPR to study protein structure and dynamics. By attaching two spin-labels, which contain unpaired electrons, to different regions of a protein, we can measure the distance between the spin-labels to gain insight of protein conformations. The signal sensitivity is limited by protons on amino acids adjacent to the spin-label. Coupling of the spin-label to these nearby protons causes decay of the EPR signal intensity. This decay is quantified by the phase memory time (T_M). While it is known that the presence of neighboring protons results in a shorter T_M, the quantitative effect these protons have on T_M is not well understood. To understand this effect, I utilize maltose-binding protein (MBP) as a model protein. I place spin-labels on various regions of MBP via mutations and site-directed spin labeling to determine how specific neighboring amino acids affect T_M. Spin-labels at MBP sites with a low number of surrounding protons are expected to have a longer T_M than spin-labels at sites with more surrounding protons. Measuring T_M values of different MBP mutants helps further our understanding of how local protein environments impact signal intensity, enabling us to develop quantitative models to predict T_M values in silico. Such models can be used to computationally determine ideal spin-label sites for proteins that previously could not be studied using EPR due to lack of signal sensitivity, in particular membrane proteins.

The modification of tRNA plays a significant role in the efficiency and accuracy of translation during protein synthesis. A modification that plays a direct role in reading cognate codons of mRNA in E. coli is the 5-oxyacetic acid methyl ester (mcmo5) modification. This modification occurs on the uracil base at position 34 (U34). The biosynthetic pathway of this modification is initiated via a hydroxylation reaction. Previous in vivo studies demonstrate the enzyme TrhP, tRNA hydroxylation protein, performs this hydroxylation reaction in anaerobic conditions. No in vitro work has been done to study this enzyme and its mechanistic function. TrhP is known to coordinate an iron-sulfur cluster, a metallic cofactor known to contribute to a variety of critical cellular processes, however, the necessity of an iron-sulfur cluster for a hydroxylation reaction is unique to this newly discovered protein family. The goal of this research project is to spectroscopically characterize TrhP’s iron-sulfur cluster to understand the importance of the FeS cluster. Site-directed mutagenesis is utilized to study the coordination of the iron-sulfur cluster. Changes to iron-sulfur cluster coordination are monitored via UVVIS, electron paramagnetic resonance (EPR), and colorimetric assays. These experiments determine how the loss of cysteine, a known iron-sulfur cluster ligand, impacts the iron-sulfur cluster coordination. Coordination of a [2Fe2S] cluster by 4 conserved cysteines is expected, and UVVIS data agrees with that hypothesis. Colorimetric assays show the cysteine to alanine mutants contain less iron than wild-type TrhP, indicating each cysteine has a significant role in cluster binding. Learning more about the specific coordination will establish the site of cluster-binding within TrhP and shed light on the cluster’s role in TrhP’s stability, geometry, and redox properties which all contribute to the enzyme’s modification activity.

Long-term neonatal intensive care unit (NICU) hospitalizations are emotionally and financially stressful for families. While family presence at the bedside and active involvement in their baby’s care have been linked to healthier outcomes, many families are unable to visit regularly. As technology continues to advance, live-streaming video platforms have been utilized in many NICUs to address the limitations of in-person presence. As the parents are able to continuously connect with their baby virtually, this allows for increased bonding with the newborn and engagement with the patient care team. The extended family and friends can interact with the infant as well. The objective of this study was to describe the utilization of the Seattle Children’s Hospital (SCH) NICU virtual visit program and its effectiveness in maintaining or increasing caregiver and patient bonding during the infant’s hospitalization. This was done through a program evaluation using mixed methodology with a parent survey and interviews. Parents and caregivers of NICU patients who used the virtual visits at least once during a patient's hospital stay were recruited using phone calls and text messages with the link to the survey. The virtual visit call frequency, time, and number of visitors were reviewed. The results showed that from July 2021 to December 2023, there were 348 virtual visits since July 2021, most of them lasting over 20 minutes (median 23.5, IQR range 3, 68.5). Most participants used the program one to two times (range min 1, max 69). Many of the calls occurred between 7 am - 7 pm with one to two attendees. In conclusion, the virtual visits program was utilized frequently and the timing of the virtual visits overlapped with the availability of the primary healthcare team which may have supported active involvement in care. The parent surveys and interviews about the program’s impact are in progress.

The University of Washington Neonatal Intensive Care Unit (NICU) Extremely Preterm (EP) program aims to provide outstanding care to EP infants born at 22 0/7-24 6/7 weeks’ gestation. The objective of this study is to assess the feasibility of night telerounds in a level IV academic NICU for EP neonates to enhance the support for teams caring for EP patients. This was a retrospective cohort study. Through review and analysis of electronic medical records, the EP database, and telemedicine records from July 2022 to June 2023, the frequency and duration of telerounds during the night shift (8-11pm) were examined. Comparisons of EP outcomes were made between two epochs: 6 months pre- and 6 months post-telerounds implementation. Descriptive statistics, chi square and independent samples t-test were used to compare EP outcomes between epochs. There were a total of 195 telerounds encounters, with improvements noted in network connectivity over time (fewer dropped calls lasting <20 sec). In the pre-implementation period, 9 (36%) of EPs were transferred for subspecialty/surgical care, mortality before 36 weeks’ postmenstrual age (PMA) was 14 (56%) and median length of stay (LOS) for survivors to discharge was 182 days (SD 72). Post-implementation, 10 (50%) EPs were transferred with mortality before 36 weeks’ PMA of 7 (35%) and median LOS for survivors to discharge was 139 days (SD 24). In conclusion, there was a reduction in neonatal mortality before 36 weeks’ postmenstrual age and a trend towards decreased length of stay during the post-implementation period. The findings suggest that night telerounds are feasible in supporting bedside NICU teams caring for critically ill EP neonates and may facilitate care advancement. Additional chart review is in progress to characterize orders entered during night rounds in order to evaluate the impact of this intervention.