3% of children live with Amblyopia (‘lazy-eye’), a visual disorder where acuity in one eye is poor even with glasses on. Amblyopia includes (1) reduced sensitivity in the amblyopic eye, and (2) interocular suppression, whereby the good eye suppresses the amblyopic eye. Our goal was to develop a method to efficiently characterize both mechanisms. Participants viewed a Gabor stimulus modulating between 0-100% contrast over time through a stereoscope, that presented a different image to each eye. Participants reported perceived contrast over time using a joystick. On each trial, the initial 14 s consisted of binocularly identical gratings modulating at 1/7 Hz, followed by 48 s where the gratings modulated at 1/8 Hz in one eye, and 1/6 Hz in the other. Separately, we measured visual acuity, stereoacuity, contrast sensitivity, the interocular suppression ratio. This method was highly efficient: only 30 min of data were needed to estimate monocular sensitivity and interocular suppression. Another advantage of this approach is that the stimulus is relatively naturalistic – the images in the two eyes are the same, except for the difference in contrast. This is important due to the nature of its binocularity and naturalistic conditions, as this will be the first of its kind - allowing clinicians a better way to assess those with amblyopia. 

Public health nurses and leaders who work for small, rural health departments (LHDs) face unique challenges that prevent the effective use of data to understand and address health equities. Among these is limited access to high-quality training in data utilization. In response, Solutions in Health Analytics for Rural Equity across the Northwest (SHARE-NW) is developing an online learning hub that contains accessible training modules for LHD leaders and practitioners to use and interpret data easier. The purpose of this study is to determine gaps in the quality of the trainings available to rural LHD practitioners in six top health priority areas: obesity, diabetes, tobacco, mental/behavioral health, violence and injury, and oral health, in order to improve population-level health equity. To date, approximately 30 training modules have been evaluated using 25 items from the Quality Standards for Training Design and Delivery tool by the Public Health Learning Network, and approximately 50 webinars have been evaluated using a set of 6 criteria. Next we will identify the patterns of gaps across the evaluation criteria and top health priority areas. Once patterns are identified, we will conduct semi-structured interviews with public health practitioners to assist in the interpretation of identified patterns. Preliminary findings suggest a lack of training activities that assist LHD leaders and practitioners to learn about data utilization in several to health priority areas. By identifying the patterns of gaps in available trainings, we aim to help busy, low-resourced rural LHDs utilize the highest quality trainings to improve their ability to interpret data and address health disparities.

Lymphatic malformations (LM) are congenital vascular malformations resulting from dysregulated growth of lymphatic vessels. LM can cause difficulty in breathing, swallowing, or eating, and in rare cases, infection or death. LM are associated with postzygotic somatic mutations in PIK3CA, which encodes for the catalytic subunit of PI3K and regulates cell growth and proliferation. Three hotspot mutations cause the majority of LM: p.E545K, p.E542K, and p.H1047R. These mutations occur at very low level in the affected tissue; usually at <10% variant allele fraction (VAF) which makes diagnosis challenging. We hypothesize that variant allele fraction (VAF) differs throughout the vascular malformation and will correlate with location in the lesion. We used droplet digital polymerase chain reaction to detect PIK3CA mutations in 8 LM subjects with 95 samples including lesion, muscle, skin, and fat. We then mapped the VAF to magnetic resonance imaging to assess genetic heterogeneity by sample location. Globally, no sample had a VAF greater than 10% indicating that assaying multiple samples does not increase the observed VAF compared to other studies where one sample was assayed. Within patients, VAF varies within the lesion, ranging from undetectable to 10%. Non-lesion tissue, including skin, and most muscle samples, had no mutation detected. All four fat samples and two muscle samples were positive for mutation with VAF <2%. We did not identify any correlations between VAF level and location within the lesion. Though we didn’t identify a correlation between VAF and location in our data, these findings will be used for further analysis including understanding how cell type composition within the samples correlates with VAF. Our goal is that this dataset will help us explore the development of LM and eventually provide information useful for the advancement of targeted therapies for future pediatric patients.

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

The Toll-interacting protein (TOLLIP) is an adaptor protein involved in the signaling pathways of interleukin-1 (IL-1) and Toll-like receptors (TLRs) in innate immunity. Evidence in published literature suggests that TOLLIP acts as a negative regulator of IL-1 and TLR-mediated immune responses by inhibiting the activity of IL-1 receptor-associated kinase (IRAK1), a serine/threonine kinase in the IL-1 and TLR signaling pathway. Lipopolysaccharide (LPS) is a major component of the gram-negative bacteria cell wall that activates host immune response upon recognition by TLR4. We hypothesize that TOLLIP deficiency leads to impaired inhibition of the innate immune response, resulting in increased inflammation in LPS-induced lung injury. We treated wild type (WT) and TOLLIP knockout (KO) mice with LPS through intratracheal instillation and bronchial alveolar lavage fluid (BALF) was collected at 3 days post-injury. Lung inflammation was measured by BALF total white blood cell (WBC) count and cell differential, BALF total protein, and BALF cytokine levels. Contrary to our hypothesis, TOLLIP deficiency was associated with decreased inflammation in LPS-induced lung injury as demonstrated by lower polymorphonuclear (PMN) cell count and significantly lower levels of cytokines in KO mice. In future studies, we will examine the mechanisms by which TOLLIP positively regulates inflammation in the LPS model of lung injury.

The generation of light at harmonic frequencies due to the non-linear interaction between a material and an electric field plays a critical role in several new technologies including ultra-short laser pulse shaping, spectroscopy, and quantum information networks. Gallium-Phosphide ring resonators can effectively convert incident light from visible to telecommunications band wavelengths, but nanoscale fabrication differences in the dimensions of ring resonators can significantly change the devices’ optimal resonant wavelengths. Unless these devices can perform optimally at uniform wavelengths, they cannot be implemented as part of any large-scale system. Post-fabrication etching methods could be used to change the width of the resonators and fine-tune their resonant frequencies. Diffusion-limited wet etching has been shown to remove angstroms-thick layers from III-V semiconductors similar to Gallium Phosphide. Hydrogen peroxide and acid solutions are alternately applied to form and remove oxide layers, reducing the width of the resonators by tens of angstroms each etching cycle. The shift in resonances near nominal wavelengths of 775 and 1550 nm after each etching cycle are measured and compared to simulated results.

The field of regenerative medicine is approaching the goal of using stem cell therapy to replace part of an organ that has been damaged irreversibly. Our laboratory differentiates kidney organoids, 3D multicellular structures that functionally and compositionally resemble the respective organ they model, from human iPSCs (induced pluripotent stem cells). However, organoids we work with are largely avascular, whilst organs in vivo are highly vascularized. Our goal for this project is to build a microfluidic, vascular platform in which organoids can grow. To accomplish this, we adopted a microfluidic chip which was fabricated using soft lithography. Consequently, PDMS (polydimethylsiloxane), a polymer commonly used in soft lithography, was molded and bound to a glass coverslip using plasma binding. With this platform, we successfully engineered microvascular networks through vasculogenesis and angiogenesis and optimized the protocol of vascularization to sustain the cells by submerging the microfluidic chip in cell culture medium. Human umbilical vein endothelial cells and human lung fibroblasts were suspended in fibrinogen ECM (extracellular matrix), seeded into the microfluidic chips with micropillars to contain the cells within their respective channels, and developed into 3D microvascular networks with visible lumen. We then stained the vasculature with endothelial cell markers (i.e. CD31, CD54, VWF) and tested the perfusability by flowing polysterene beads through the microfluidic chip, observing the retention of polysterene beads within the vessels. Finally, we altered our design, specifically the height and width of the channels, to incorporate kidney organoids. Currently, we are using this platform for vascularizing kidney organoids and simultaneously implementing a flow system to induce shear stress on the microvasculature to attain physiological parameters. Ultimately, we aim to vascularize a kidney organoid to demonstrate the vascularization of stem cell tissue in vitro and see growth of tissue within our system, which would further our process in the translation pathway from bench to bedside for kidney regenerative medicine.

Nitrogen-vacancy centers (NVs) are defects in diamond whose spin and optical properties enable their use as qubits, the basic building blocks of quantum information processing (QIP). Our group is interested in forming NV centers with good optical properties near the surface of a diamond substrate to create a scalable QIP platform. To form these near-surface NV centers, we implant nitrogen ions into a diamond and subsequently anneal. Recent published literature suggests NV centers formed by implantation and annealing protocols have poor optical properties, including optical linewidth and spectral diffusion, while NV centers formed from naturally-grown-in nitrogen have good optical properties. In this project, we examine this discrepancy by studying the optical properties of NVs formed from implanted nitrogen (¹⁵N isotope) versus grown-in nitrogen (¹⁴N isotope). We implant diamonds with ¹⁵N ions, anneal to form NV centers, then identify each NV center’s nitrogen isotope using optically-detected magnetic resonance (ODMR) to establish the NV’s origin (from implanted or naturally occurring nitrogen.) We then measure the optical properties of the NV centers using photoluminescence excitation (PLE). Preliminary results show a statistical correlation between good optical properties and ¹⁴N NV centers. In this project, I aim to analyze PLE data to measure NV optical properties, use ODMR to identify NV isotopes, and correlate these two measurements for the same NV centers. This work will identify the source and study the spectral properties of implanted NV centers in diamond, ultimately assisting in the development of a scalable QIP platform.

Rheumatoid Arthritis (RA) is a chronic disease with no known cure. While medications are often effective at managing physical symptoms, RA patients often experience high levels of fatigue. Studies have found that fatigue may be managed through meditation, but little is known about virtual reality meditation’s (VRM) potential to alleviate fatigue. The purpose of this study is to examine the feasibility and acceptability of VRM as an alternative non-pharmacologic intervention for fatigue management in RA patients. This study implements a convergent mixed-methods design to collect patient feedback. Four participants diagnosed with RA were recruited from a local rheumatology clinic. Participants used a VRM headset in their own home over the course of four consecutive weeks. During this time, Patient Reported Outcome Measurement Information System (PROMIS) measures of fatigue, pain, depression, anxiety, physical activity, and mood were taken at baseline and at weekly intervals. Semi-structured interviews occurred at baseline and at the conclusion of the study. Interviews were audio recorded, transcribed, and coded using Atlas.ti (v8). The results are currently pending. Expected results include that participants will find VRM both feasible and acceptable for fatigue management, and that participants will report reduced fatigue levels after using the VR device. While studies have explored the use of VRM in the treatment of anxiety disorder, depression or PTSD, this is the first study to examine VRM’s use for managing fatigue in participants with RA. Results of this study will inform future clinical trials using VRM, implementation of VRM into clinical use, and give a better understanding of the patient’s experience of utilizing VRM for fatigue management. 

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