Brain stimulation has emerged as a novel treatment for stroke, a prevalent cause of death and disability worldwide. Studies in rodent models have shown that post-stroke electrical stimulation results in plasticity and neuroprotective benefits. However, techniques that were effective in rodents have rarely translated into clinically viable therapies in humans due to the significant differences in rodent and human neurophysiology and anatomy. Therefore, the goal of our study is to obtain clinically relevant outcomes that describe the mechanisms of stimulation induced plasticity in non-human primates. We combined electrophysiology and immunohistochemistry to investigate the degree of stimulation-induced plasticity and network dynamics after photothrombotic stroke in 4 macaques. We quantified the expression of two biomarkers, postsynaptic density-95 (PSD-95) and growth-associated protein-43 (GAP-43) in cells within ~10mm from the lesion penumbra. Since PSD-95 is important for the maturation of excitatory synapses and GAP-43 is involved in axonal branching and elongation, evaluating the expression of these two proteins around the lesion core allowed us to compare post-stroke synaptic and axonal plasticity in 2 control and 2 stimulated monkeys. Based on wide-field epifluorescence imaging, we identified the distance from the lesion penumbra at which there was a distinct difference in biomarker immunoreactivity in control and stimulated animals, and performed high-magnification confocal imaging to further investigate the structure of biomarker expression. Furthermore, analysis of the electrocorticography signal showed a largescale downregulation of neural activity following electrical stimulation, while Nissl staining revealed that stimulated monkeys had smaller lesion volumes than controls. These results indicate that stimulation elicits changes at both neurophysiological and cellular level, and may exert a neuroprotective effect on the post-stroke network by reducing metabolic energy consumption.. Therefore, this study investigates the effects of electrical stimulation on neuroplasticity and protection following injury, which may have a profound impact on future therapeutic interventions for stroke.

The gut microbiota, or the microorganisms that live in the gastrointestinal tract (GI), play a significant role in both gut and overall health. In LMICs (Low-Middle Income Countries), diarrheal diseases are among the leading causes of death in children under five. The ongoing EcoMiD (Enteropatógenos, Crecimiento, Microbioma, y Diarrea) project aims to study the interaction between infants' gut microbiomes and viral, bacterial, and parasitic enteric pathogens along a rural-urban gradient in Ecuador through a cohort study of 600 mother-child dyads. The objective of this portion of the study was to validate a method for assaying multiple enteric pathogens in stool samples simultaneously using TaqMan Array Cards (TAC). Before samples can be analyzed using TAC, each of the 56 pathogenic targets of interest must be validated to determine the limit of detection, limit of quantification, inhibition effect of the stool matrix on detection, repeatability and reproducibility. The TAC Positive Control validation process initially involves growing up each of the organisms of interest, extracting its' total nucleic acids (TNA) and quantifying this through (RT)-qPCR and Qubit. Analysis of this data enables seeding the positive controls at known concentrations to quantify target TNA for TAC. Of the 56 targets, we have completed this work with 35 targets (26 reference organisms, 9 gBlocks). For targets that are time/cost intensive to work with or not culturable, gBlocks are utilized in place of reference organisms. gBlocks are synthetic gene fragments with a known sequence. Next steps of this process includes validation of the 21 remaining pathogens, some of which involves working with anaerobic organisms. Analysis of this data will inform seeding concentrations to validate the developed TAC and enable quantification of these targets in stool samples from Ecuador. 

Individuals experiencing homelessness and housing instability in King County have inadequate access to water, sanitation, and hygiene (WASH) services. This urgent matter of community health has only been exacerbated by the coronavirus disease (COVID-19) pandemic in recent years and can necessitate open defecation or unsafe disposal of wastewater. Particularly among those residing in recreational vehicles (RVs) and use their bathroom facilities, unsafe sanitation practices increase the risks of intestinal pathogen transmission and infection in densely populated communities. In a continuation of efforts to address this, I have developed a study that aims to identify an optimal method of RV wastewater concentration for downstream detection of pathogens. With informed consent from RV residents and sampling assistance from Seattle Public Utilities, I have began collecting a representative collection of samples of RV wastewater from multiple neighborhoods across Seattle. I have utilized a split-and-seed approach in which half the samples were seeded with known amounts of target bacterial, viral, and spore-forming organisms, and the other half left unseeded as a control. These samples will be used to compare two methods of sample concentration, skimmed milk flocculation and membrane filtration, using a weighted rubric that evaluates biosafety, seeded organism recovery efficiency, personnel time, and cost. Upon observation in the lab, I have found that skimmed milk flocculation led to inhibition during detection via molecular assays. Additionally, high turbidity of the samples yielded significant logistical challenges with processing skimmed milk samples, leading my team and I to instead favor the membrane filtration technique. As we move forward, I anticipate detecting elevated levels of pathogens in wastewater from RVs when compared to samples from control locations chosen to represent populations living in sewered and unsewered environments representing populations. Upon development of the optimized protocol, my work will be prepared for publication in a peer-reviewed journal and shared with the City of Seattle.

Creating novel anti-cancer drugs aimed at targeted glioblastoma multiform (GBM), a type of brain cancer, is slow, extremely expensive, and remains a persistent challenge within the medical field. To address this challenge, our team’s research project is aimed at creating scaffolds from chitosan-hyaluronic acid (CHA) to mimic the brain microenvironment and serve as a platform for high throughput screening (HTS) of cancer drugs. 3D culture systems can promote more cell-cell and cell-matrix interactions, which can closely mimic the in vivo extracellular matrix environment. Studies have shown that the drug resistance of 3D-cultured cancer cells can better reflect the in vivo situation, and thus can potentially improve the success rate in drug screening processes. CHA scaffolds are especially beneficial for culturing GBM cells, as hyaluronic acid (HA) is a major component in brain tissues. To generate the scaffolds, we used different freezing rates and temperatures to create freeze-dried 8 wt% CHA scaffolds with pore sizes of 60, 120 and 180 μm. We characterized the compressive modulus of the scaffolds using the Instron test machine, and the porosity using liquid replacement methods. Cell studies with 3 different cell lines are currently being conducted on these scaffolds, after which an AlamarBlue assay will be used to determine the optimal pore size for each cell line in terms of their growth and drug resistance. The results of this can prove that our CHA scaffolds have good flexibility in response to different cancer cell line 3D cultures and have good potential to be an HTS platform.

Measurement based care (MBC) is an evidence-based approach to psychological care that has been gaining momentum in the field. MBC emphasizes the use of frequent, repeated administration of measures and the consequently collected data in order to inform the course of treatment. This contrasts with traditional care, which relies more heavily upon clinical judgment and only sparse use of measurements. MBC has been shown to provide significant improvements for patients, largely due to increased patient feedback which results in quicker responses in changing treatment plans accordingly. Ecological momentary assessment (EMA) is a data collection strategy where researchers repeatedly remotely administer surveys that ask about participants’ current real-time states, and can provide additional clinical utility to MBC treatment through more contemporaneous data which is less subject to the errors of retrospective recall. Despite EMA’s ability to gather more ecologically valid data, there is a lack of research on how to optimize EMA to reduce concerns of response burden and adherence. This research aims to help clarify this knowledge area by examining how timing and frequency of measures affects participants’ adherence to an EMA procedure. We recruited 215 participants from the University of Washington’s Psychology Subject Pool, and randomly assigned each participant to one of two groups—each group was tasked with answering the same short survey, a modified 5-item version of the Subjective Happiness Scale (SHS), either three or seven times a day, both over a period of three weeks. We predicted that there would not be a significant difference in adherence rates between the two different sampling frequencies, and that surveys taken earlier in the day would be completed less frequently than those in the evening. Understanding participants’ responses to EMA is critical not only for optimizing future psychological research, but also for improving the use of measures in clinical care.