The composition of metropolitan governance has many effects on land use decisions, budget allocations, housing development, transportation planning, and racial, economic, and social equity in urban areas. However, there has been little academic inquiry into the effect of regional governance structure on transportation accessibility. This paper seeks to examine statistical linkages between regional governance fragmentation and trends toward and away from greater transportation accessibility in metropolitan areas. I perform a comparative statistical analysis of 47 of the 50 largest Metropolitan Statistical Areas, examining census data from 2002 to 2022 and transit accessibility data from the University of Minnesota Accessibility Observatory from 2014 to 2021 to examine this relationship. The causal factor I investigate is metropolitan governance fragmentation, which I capture through a Governance Fragmentation Index (GFI). The dependent variable, transportation accessibility, is captured through an Accessibility Gap Index, which categorizes transportation access through accessibility levels throughout each Metropolitan Statistical Area, utilizing data from the Accessibility Observatory from 2014 to 2021. My analysis controls for potential confounding variables, such as geographic area, population size, poverty levels, and region. I expect to find that lower levels of governance fragmentation in a Metropolitan Statistical Area will be associated with greater gains in transportation accessibility. Whether or not a significant relationship is identified, the research conducted will contribute to literature and ongoing research surrounding metropolitan governance and transportation accessibility.

Lipotoxicity in cells occurs when lipids in biological tissue accrete to a toxic level. This toxic buildup is associated with obesity and type 2 diabetes, two of the leading causes of death around the world. Various studies have induced obesity in mice through a diet consisting of high levels of saturated fats. Some of these studies also investigated the effects Adefovir Dipivoxil, an antiviral and inhibitor of mitochondrial DNA replication, had on these obese mice. My research investigates fibroblasts that have been grown in a high-lipid environment and how Adefovir Dipivoxil affects these cells. We hypothesized that administering Adefovir to these cells would halt apoptosis and prevent further progression of lipotoxicity by stabilizing levels of triacylglycerol synthesis. To test this, I induced lipotoxicity in the cells using palmitic acid, a saturated fatty acid. Next, I treated the cells with Adefovir Dipivoxil. To measure the efficacy of the drug, I used a staining kit to measure the ratio of live to dead cells before and after administering the drug. Furthermore, I used a dye-based assay to measure the intracellular triglyceride levels before and after treatment. In addition to treating fibroblasts with palmitic acid, I treated a new set of fibroblasts with oleic acid, an unsaturated fatty acid, to determine how Adefovir acts on these cells. Data from this research will contribute to further understanding the mechanism of lipotoxicity on various cells, as well as the role that mediators of mitochondrial function like Adefovir Dipivoxil could play in treating lipotoxicity. On a broader scale, we hope that this research will provide insight into future treatments for obesity and type 2 diabetes.

Birth asphyxia is the inability of a newborn to begin and maintain breathing. Twenty-three percent of neonatal deaths globally are caused by birth asphyxia. Birth asphyxia results in a neurological injury called hypoxic ischemic encephalopathy (HIE). Rapid HIE screening within six hours after birth is crucial to identify neonates at risk. Unfortunately, the diagnostic equipment is impractical for low resource settings because it is costly ($20/test and $5,000 for equipment) and requires technical staff, that are in short supply, to operate. We hypothesize that a cost-effective device can be developed for HIE analysis. pHast Cam quickly screens for birth asphyxia and HIE in infants via a paper-based blood pH sensor. The device combines an inexpensive pH sensitive dye, a smartphone camera, and a fixture that controls the imaging environment to quickly identify acidosis from samples. A low-cost paper-based strip is made with a water-soluble resin doped with a pH-sensitive dye, bromothymol blue (BTB), and a membrane to filter out red blood cells. The fixture removes lighting variation. The smartphone camera records the pH indicator image, and an algorithm captures, reduces noise, and accesses color change. pHast Cam incorporates four features: 1) accurate assessment of acidity within 0.05 pH units, 2) require only a few microliters of sample, 3) use electrical hardware and software only from the smartphone, and 4) affordability. At this stage, we have achieved a regressive linear model that predicts buffered solution acidity (y=-589.32x+4684.05 R2=0.9857), with 95% confidence interval of 0.04 pH units. In the future, we will transition from measuring buffered solutions to blood-plasma. Ultimately, we expect pHastCam to screen for birth asphyxia, and other acid-base disorders, by quantifying plasma pH in neonates so that timely therapeutic interventions and plans to address long-term complications may occur.

Capillary microfluidics capitalize on surface tension effects encoded in microchannel geometry and chemistry to transfer liquids without external instruments, making them a user-friendly technology for point-of-care tests. For most applications, hydrophilic surfaces (contact angle < 90Ëš) are necessary to induce surface tension driven flow. Currently, this is achieved with vacuum plasma chambers that alter surface chemistry. Unfortunately, the hydrophilic properties made with plasma processing are temporary and unstable. Alternatively, an inherently stable hydrophilic 3D-printing resin containing polyethylene glycol diacrylate (PEGDA) and acrylic acid (AA) was recently developed for capillary microfluidics. However, this hydrophilic resin has not been thoroughly validated for inexpensive (<$300) liquid crystal display (LCD) printers. Our objective is to optimize and validate 3D-printing parameters including exposure time, UV power, layer thickness, and lift/retract speed using this hydrophilic PEGDA-AA resin with three LCD 3D printers (AnyCubic Photon Mono X 6K, AnyCubic Photon Mono M5s Pro, and Phrozen Sonic Mini 8K). Validation includes measuring hydrophilic properties as well as the dimensional fidelity of the printed channels compared to the design specifications. Our proof-of-concept prints on the Mono X 6K printer had average contact angle measurements of 42.8° ± 8.77. The percent differences between designed and printed channel lengths, widths, and depths were 31.5 ± 0.23%, 28.9 ± 3.41%, and 2.40 ± 13.9% respectively. By optimizing the print parameters of cost-effective 3D printers with the inherently stable hydrophilic resin, we enable capillary microfluidic technologies for users in low income/resource settings who may not have access to vacuum plasma chambers. Future work will explore additional resin modifications to encourage applications like spatial patterning of hydrophilicity and protein immobilization in microchips. [1]V. Karamzadeh, A. S. Kashani, M. Shen, and D. Juncker, “Digital Manufacturing of Functional Ready‐to‐Use Microfluidic Systems,” Advanced Materials, vol. 35, no. 47

Tissues like the meniscus, a wedge-shaped pad of connective tissue found in the knee, are fibrous and have complex architecture that regenerates poorly and undergoes active mechanical stimulation which modifies cell signaling and tissue health. In vitro models are beneficial for characterizing these interactions as they create a controlled environment where single variables can be altered. We previously used the J1 Mechanoculture bioreactor to apply strain on a fibrous polymer scaffold laden with primary meniscal cells and observed nonsignificant variances between testing groups with mock injury vs. no injury. Applied strain was modeled after physiological strain levels, ~10%. Based on the minimal changes in cell behavior observed in mock injury samples, it is likely that the mock injuries in conjunction with the applied strain did not induce comparable plastic deformation to that experienced post injury within the native meniscus. We hypothesize that increasing strain and applied force to achieve plastic deformation within the electrospun samples will create a fibrotic and apoptotic response like that in vivo. Ongoing work is analyzing how the bioreactor will interact with unaligned electrospun polymer samples with no cells present. This will demonstrate the optimal parameters to instigate a significant material response. By inducing significant changes to scaffold material properties and underlying structure, it is more likely cells with demonstrate fibrotic and apoptotic responses in vitro mimicking immediate cell reactions to meniscal injuries in vivo. This response will be assessed by assaying for fibrosis through αSMA activation and apoptosis by caspase-3 activation. On the conclusion of this study, we expect that greater applied stress and associated strain will cause more plastic deformation within the polymer scaffold. This can be applied to an in vitro meniscus injury model to better understand the response of primary meniscal cells to stress in an environment with disrupted mechanics.

Alzheimer's Disease (AD) is clinically characterized as a predominantly amnestic (memory impairment) syndrome at presentation that progresses to affect other cognitive domains. AD is pathologically defined by the presence of amyloid plaques and neurofibrillary tangles of hyperphosphorylated tau (pTau) in stereotypical brain regions. AD shows clinical and pathological diversity, including non-amnestic subtypes, severity of tau pathology across brain regions, and co-pathologies such as aggregates of hyper-phosphorylated transactive response DNA-binding protein 43 (pTDP-43). This study aims to examine the association between pTau and pTDP-43 using new highly quantitative approaches. By examining the combined pathology, we hope to identify patterns of pTau related to pTDP-43 across the different clinical and pathologic subtypes. The University of Washington Alzheimer's Disease Research Center clinical core autopsy cohort was characterized and subdivided into amnestic and non-amnestic syndrome subtypes. The subjects were analyzed to identify the prevalence of pTDP-43 and its correlation to the subject's cognitive data and patterns of progression. This analysis was used to select a subset of 29 cases with non-amnestic dementia and a matched subset with an amnestic subtype for more in-depth neuropathological and molecular profiling of several brain regions. Using the HALO platform, I generated quantitative measures of pTau in the frontal, temporal, and parietal cortex, as well as the hippocampus. The integration of these findings aims to understand how pTDP-43 pathology influences tau distribution based on clinical presentation These results will allow us to select a small set of cases for further work that will include using NanoString GeoMx Digital Spatial Profiling to identify potential pathways relevant to the association between pTDP-43 and pTau severity concerning mechanisms of clinical and pathologic heterogeneity in AD. These insights will allow for further research of these pathways to determine their biological relevance and ways to mitigate their effects.

Alzheimer’s disease (AD) is the most common cause of dementia in the aging population, characterized pathologically by the presence of amyloid plaques and tau neurofibrillary tangles in the brain. However, AD often coexists with other pathologies contributing to dementia, such as hyperphosphorylated aggregates of the protein TDP-43. TDP-43 induces a dementia syndrome similar to AD and the combination of AD and TDP-43 is associated with accelerated cognitive decline, greater brain atrophy, and increased AD pathologic burden. AD and TDP-43 pathology are definitively diagnosed post-mortem upon neuropathologic examination but there is a great need to be able to identify these pathologies in living patients using biomarkers. Currently there are accepted biomarkers for AD, including measures of amyloid beta and hyperphosphorylated tau proteins in cerebrospinal fluid (CSF), but there are no biomarkers for TDP-43. Leveraging the reliability of CSF in detecting pathologic proteins, we hypothesize that measurable hallmarks of underlying TDP-43 pathology also exist in CSF. We tested four groups of brain donors (n=36 per group) defined by presence or absence of AD and TDP-43 pathology at autopsy: healthy controls, AD only (amyloid plaques and tau tangles), TDP-43 only, and AD+TDP-43. Post-mortem CSF samples are analyzed for TDP-43, hyperphosphorylated tau (pTau-181), and the brain injury marker glial fibrillary acidic protein (GFAP) using the Quanterix SR-XTM Biomarker Detection System. Because these assays are intended for ante-mortem samples, the first aim of the study was to determine optimal sample preparation for post-mortem samples, followed by the second aim to determine if there are concentration differences between proteins in CSF across groups. Successful identification of reliable TDP-43 biomarkers in living patients would improve neurodegenerative disease diagnostics, enabling accurate underlying pathology diagnosis and facilitating tracking disease progression and treatment response as therapies for AD, TDP-43, and other causes of dementia emerge.

According to the World Health Organization, Alzheimer’s Disease (AD) is the most common form of dementia – a major and growing cause of disability and dependency among older people globally. The Seattle AD Brain Cell Atlas (SEA-AD) project is a collaboration between the University of Washington (UW) and the Allen Institute for Brain Science (AIBS) aimed at discovering early vulnerable cell types in AD. In SEA-AD, we hope to further our understanding of the etiology and early progression of AD to ultimately identify targets for effective therapeutic intervention. Eighty-four participant brain donors with a postmortem interval less than 12 hours from the UW AD Research Center (12/84) and Kaiser Adult Changes in Thought (72/84) studies were included in the SEA-AD cohort. At the time of procurement, one hemisphere was frozen in super-cooled isopentane for transcriptomic analysis at AIBS; the contralateral hemisphere was fixed in 10% neutral buffered formalin for neuropathologic assessment at UW. The middle temporal gyrus, medial entorhinal cortex, and hippocampus were sampled, processed, embedded in paraffin, and sectioned for  immunohistochemical (IHC) studies. Seven antibodies, including duplexed stains, targeting amyloid b (6e10) and microglia (IBA1), pTau (AT8) and pTDP-43 (1D3), monoplexed a-synuclein (LB509), astrocytes (GFAP), neurons (NeuN), and triplexed histochemical stain: hematoxylin, eosin, and Luxol fast blue were deployed to assess the neuropathology associated with the presence and progression of AD and related neuropathologic changes. The data obtained from the quantitative assessment of the IHC staining is integrated with the transcriptomic data generated by the Allen Institute to enhance our understanding of the cellular vulnerabilities and associated molecular processes of AD. Public access to this neuropathological data through the SEA-AD resource potentiates research efforts to understand and identify the mechanisms of AD progression.

It is well documented that women are predisposed to various musculoskeletal injuries, including hip labrum, Anterior Cruciate Ligament (ACL), and meniscus tears. Literature has detailed the regenerative potential of human mesenchymal stromal cells (hMSCs), and clinical trials have confirmed their medicinal application within tissue repair and healing, particularly in the musculoskeletal system. hMSCs are commonly used in the field of tissue engineering due to their immunomodulatory capabilities, differentiation capacity, and autograft availability. Across the lifespan, cells in male and female bodies experience varying levels of estrogen exposure, which elicits different responses. During in vitro cell culture, cells are typically exposed to sources of exogenous estrogens, including phenol red and fetal bovine serum (FBS). Although these additives are commonplace in the practice of cell culture, little research has been done to understand the impact of these additives on hMSCs in in vitro cultures, especially among female hMSCs. To investigate these cellular responses, we studied the impact of these estrogen-mimetic media components on cell proliferation and metabolism in vitro for hMSCs derived from male and female donors. Specifically, we investigated phenol red, which is shown to behave as an estrogen, and FBS, which naturally contains 17β-estradiol (E2). We hypothesized that estrogen-mimetic compounds would be associated with an increase in cellular proliferation and metabolism in a sex-dependent manner. These results clarify the response patterns of male and female hMSCs due to exogenous estrogen exposure, improving our sex-specific understanding of their potency for in vitro studies and regenerative medicine applications.