Sex trafficking, the practice of transporting and coercing persons into commercial sexual exploitation, is a global problem that’s supported by our actions (or lack thereof) here in the U.S. The question this report seeks to answer is how international sex trafficking continues to be supported in the U.S. and in Washington State in particular. Sex trafficking is a difficult topic to research, as most data lies in illegal activities among hidden populations. If, however, people are able to inform themselves about the impact of sex trafficking on the sex industry in the U.S., they may demand reform on both a societal and governmental level to combat sex slavery. My prediction is that a large factor contributing to sex trafficking’s success is simply the public’s lack of knowledge on the subject. Additionally, it’s likely that laws and societal norms as well as lack of resources/funding for anti-trafficking efforts are also contributing factors. To answer my research question I’ve read through various encyclopedias, scholarly journals, organization websites, and even a report on human trafficking in Washington State. I’ve also conducted an interview with a survivor of sex trafficking who now volunteers with helping victims of trafficking. My research focuses on development of sex trafficking, who supports it, why it thrives in Seattle, and what needs to be done to stop it. Research shows that sex trafficking is prominent in Washington State due to location, proximity to the border, and client supply. Changes in societal attitudes in conjunction with revised laws that better protect victims and punish the traffickers and greater support/funding for organizations against sex trafficking will all serve to combat this problem. If the world decided to stop human sex trafficking, then millions of people could be spared from a very tragic fate.

Foraging behavior in mammals requires the integration of various sensory cues to allow the subject to navigate through the environment. When the search becomes predictable and repetitive, memory can be used to enhance the sensory cues. To model the dynamic interactions between sensory and memory, we performed a comparative analysis on rats and humans by designing a two-stage task paradigm: search and return. The subjects were trained to search for rewards in a confined space with limited sensory cues. After the end of each trial, they were trained to return to the start position. The rat task was performed in an automated arena that allowed the rats to follow the reward’s olfactory cues but prevented visual and hearing cues. Similarly, the human task simulated the paradigm using a computer game that allowed the subjects to search and return in a virtual room with limited visual cues. We predict that sensory cues have a more significant impact on the search strategies compared to the return strategies. If the reward locations are fixated in every trial, we also expect the subjects to increasingly rely on memory for both the search and return strategies. Our research will provide insight on the tracking methods and analysis in studying the roles of sensory and memory in complex foraging behavior.

Many 2 dimensional materials, such as Tungsten Diselenide (WSe₂) have many interesting device applications such as thin and flexible photo detectors and transistors. We use a process called chemical vapor deposition, which involves depositing a vapor of WSe₂ on a substrate by a specific application of heat. Specifically, we installed a secondary heating coil in our furnace which allows for a region of uniform temperature near the substrate. This modification allows for large scale growth of single crystals on the substrate. This approach to manufacturing has the advantage of allowing more samples to be studied and inevitably being more applicable to industrial applications. To determine if this method is feasible, we probe the quality of these crystals by measuring their electrical properties.

The pharmaceutical industry’s “one-size-fits-all” approach to drug discovery, coupled with the use of animal models that do not maximize resources or accurately represent human disease, is a significant obstacle to the development of more effective treatments with the capacity to significantly improve patient wellbeing. The concept of personalized medication needs to be prioritized within the drug development process to advance the identification of drug candidates with the greatest potential to ameliorate patient-specific symptomatic progression. This is especially true in neuromuscular diseases like Duchenne Muscular Dystrophy and X-Linked Myotubular Myopathy, which show wide variability in severity and age of onset. Myogenesis is the formation of muscular tissue, specifically during embryonic development. The involvement of chemical cues to promote the fusion of myoblasts into multinucleated structures called myotubes is fairly well understood. However, the specific mechanical forces associated are yet to be characterized due to the complex cellular machinery at play. Recent evidence suggests that exposing muscle cells in culture to rehabilitation techniques, such as mechanical and electrical stimulation that mimic resistance exercise, has profound functional consequences. Therefore, I hypothesize that subjecting human skeletal muscle myoblasts (HSMM) to mechanical stretch will improve maturation in vitro and manifest a phenotype that mimics muscle tissue. To test this hypothesis, experiments are currently underway using the STREX Cell Stretching System to examine the various parameters that invoke differentiation of these HSMM cells. Varying cyclical strain frequencies and ratios along with durations of stretch and rest that mimic static/dynamic stretch cues from rehabilitation therapy are being used to determine the ideal stretch criteria for muscle cell growth and differentiation. This stretch protocol will eventually provide guidance toward the production of skeletal tissue engineered products in vitro to be used for drug development to promote the recovery of muscle function after injury and possibly forestall disease progression.

Ion Mobility Mass Spectrometry (IMMS) has become an important tool for separating small molecules such as peptides, glycans, and petroleum molecules. Ion Mobility is most often performed in He, N₂ or air. There is a great interest in using alternative drift gases to increase selectivity of ion mobility separations, specifically for complex mixtures like petroleum. In this study we measured arrival time distributions (ATD) of a set of double bond equivalents (DBE) in four different gases (He, N_2, CO₂, Ar) which span a range of masses and polarizabilities. Quinoline, isoquinoline, and their DBE were chosen because they are aromatic compounds that are easily protonated due to the presence of nitrogen and are similar to molecules found in petroleum. The measured ATDs were plotted as a function of reciprocal drift voltage; the resulting slope is inversely proportional to mobility. Using the mobility and the Mason-Schamp equation, we determined the collision cross section of each molecule with each drift gas. We also investigated the relationship between the dipole moment of an ion and its collision cross section. In general, collision cross sections increase with drift gas size and polarizability. Additionally some of the DBE were better separated with larger, more polarizable drift gases (N₂ and CO₂). We have shown that IMMS can be used with different gases to improve separation between ions of similar size and shape. This method can be used in future investigations of petroleum mixtures to potentially identify a wider range of molecules.

Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutation in either of the genes PKD1 or PKD2. This disease has been the focus of ongoing research due to its high prevalence and lack of curative treatments. How does ADPKD develop? To understand cyst formation in ADPKD, we are utilizing the murine model PKD1 p.R3277C (RC). Human dominant expression of the APKD phenotype is not similarly found in mice; null mutations of PKD1 or PKD2 are embryonic lethal, and the cystic phenotype of mice is observed only in conditional deletion mutants. However, the RC murine model recapitulates the late onset cystic growth of human ADPKD. The morphology of the pre-cystic kidney was characterized by formalin-fixed paraffin embedded (FFPE) sectioning in combination with H & E staining performed on RC homozygous mutant and control kidneys at post-natal time points of p20 (days), p40, p60, and p80. In the future, RNA from RC mutant and wild-type kidneys at p20 will be collected and sequenced. RNA sequencing will be utilized to determine pre-cystic expression changes between affected and unaffected phenotypes. Although less advanced, cyst formation has been shown to occur as early as p20. Because of this, the earlier time point of p10 was also assessed. By investigating these time-points we aim to identify the earliest differences that we hypothesize are more likely to be causal for cyst formation, versus changes that are a secondary consequence of PKD pathology.

Approximately 300,000 cases of appendicitis are diagnosed in the United States on a yearly basis. Appendicitis is the condition in which the appendix, located in the right lower quadrant (RLQ) of the abdomen, becomes inflamed usually due to a bacterial infection. A common symptom of appendicitis is when RLQ pain is sustained over time. We, the Surgical Outcomes Research Center (SORCE), hypothesize that antibiotics for intra-abdominal infections recommended by the Infectious Diseases Society of America and American Surgical Association are just as effective as appendectomies for treatment. This project will illustrate to both clinicians and patients that they will have more options when treating appendicitis. The Comparing Outcomes of Drugs and Appendectomy (CODA) clinical trial aims to investigate appendicitis treatment options and understand if antibiotics are just as good as appendectomies (the current standard of care) from the patients' and clinicians' perspectives. Previous studies in Europe suggest the usage of antibiotics is a viable option when treating appendicitis. Patients consenting to the CODA trial are randomized to determine which treatment they receive, either the surgical appendectomy or the antibiotic medication. If the patient wishes to not be randomized, they can opt-into an observational or electronic medical record cohort in which they choose their treatment option. We are currently still in the data collection stage as we continue to monitor and assess patient reported and clinical outcomes for two-years following initial presentation. This research is important because patient reported outcomes gained from the trial will educate patients about their treatment options, therefore patients can decide the treatment best suited for them. In addition, the same results may demonstrate to current healthcare professionals that appendicitis may be treated though other methods, namely antibiotics, which if successful, will revolutionize current perspectives on how appendicitis is treated in the United States.

Urban development is occurring at an unprecedented rate in the city of Seattle, WA. What were once healthy salmon spawning streams are now highly altered ecosystems that reflect the impacts of human land-use change. However, little work has been done on the impact that these changes are having on the carbon biogeochemistry of urban streams. We investigated the biogeochemical properties over time of Thornton Creek, a recently daylighted and restored stream network in Seattle. We conducted longitudinal sampling at three locations along Thornton Creek in a unique and newly restored area. Our longitudinal study consisted of weekly headspace and chemistry sampling in addition to data collected continuously from carbon, dissolved oxygen, and temperature sensors installed in the stream. We used the headspace method to analyze dissolved gas concentrations contained in the stream water by equilibrating the stream water with atmospheric air and injecting the samples into evacuated exetainers. The samples were then run through a GC-MS machine and corrected using atmospheric samples to reveal the dissolved stream carbon dioxide and methane concentrations. The results of this research have implications for how humans have had an impact on the concentrations of dissolved gases in the surrounding water bodies with ramifications for urban streams as localized sources of carbon dioxide and methane to the atmosphere. Additional data on water chemistry highlights the consistency of these gas concentrations over time.
 

During development, the repression of transcription factor tbx16 allows embryos to dynamically allocate neural and muscle tissue in the correction proportion as the body axis is formed. Understanding how tbx16 is regulated is crucial for identifying the underlying mechanisms of neuromuscular developmental defects. Currently, we are investigating how two genes of interest termed eve1 and vent interact to regulate the expression of tbx16 in zebrafish embryogenesis. Previous studies have relied on Morpholino anti-sense oligonucleotides (MOs), but MOs often create off-target effects and misleading phenotypes that don't reflect genetic mutants. Using the CRISPR-Cas9 system in zebrafish, we have created true genetic knockouts for eve1 and vent. By analyzing the difference in phenotypes between eve1 mutants, vent mutants, and eve 1/vent double mutants, we can determine if eve1 and vent act redundantly to repress tbx16. Our goal is to uncover the mechanisms behind these mutant phenotypes in order to identify the role that eve1 and vent play during embryogenesis. Our research aims to further our understanding about how serious birth deficits arise from gene deficiencies and complications in development.

Photovoltaic devices based on polycrystalline films of hybrid organic-inorganic lead perovskite semiconductors have demonstrated impressive power conversion efficiencies of over 22% while still retaining the advantage of low-cost solution processability. However, a fundamental understanding of many of the photophysical and charge transport properties of these materials must be further developed. In this project, we synthesize a series of n-type doped single crystals of organic-inorganic methylammonium lead bromide (CH₃NH₃Pb_1-xBi_xBr₃) and all-inorganic cesium lead bromide (CsPb_1-xBi_xBr₃) by varying the amounts of Bi3+ cation and use optical spectroscopy and x-ray diffraction to characterize their optical and structural properties. In collaboration with the Chu group in the UW Department of Physics, we use these crystals to compare the charge transport properties of organic-inorganic to all-inorganic perovskite as a function of temperature and doping concentration, in order to understand the mechanisms that determine the charge carrier mobility in these materials. Insight gained from studies on these crystals can be applied to improve device fabrication procedures, ultimately improving perovskite device performance and stability and thereby hastening the introduction of efficient perovskite-based materials into the emerging solar energy market.

Small cell lung tumor (SCLC) is a type of lung tumor that accounts for 10-15% of new lung cancer cases. It is an aggressive cancer that is often not detected before metastasis occurs. Even with chemotherapy and radiotherapy, SCLC has a very high mortality rate and patients tend to develop chemoresistance quickly. Unlike other types of lung cancers, SCLC has no available gene-targeted therapies. This project examines the development of chemoresistance in SCLC genetic mouse models . This approach deletes tumor suppressor genes, such as Rb1 and Tp53, and/or activates oncogenes like lmyc, using a Cre-expressing virus in mouse lung tissue, which leads to the development of SCLC. My role included verifying tumor burden following magnetic resonance imaging (MRI) and quantification of tumor volume in the lungs using MRI data. Once MRI confirmed tumor burden, mice are treated with chemotherapy and tumor volume measured over three weeks of drug treatment. The tumor response to chemotherapy will be compared in different genetic mouse models. At the end of treatment, lung tumors are collected and I use immunohistochemistry to measure the response of the tumors collected from the genetic mouse models. I examine markers of cell proliferation and cell death to assess drug effectiveness. These results could be relevant clinically if certain genetic mutations are found to be associated with chemoresistance.

Hair cells are mechanosensory receptors of the auditory and vestibular systems in vertebrates. Loss of hair cells or hair cell dysfunction, leads to defects in hearing and balance. There are several mitochondrial gene mutations that lead to hearing loss, however, whether these mutations affect hair cells directly remains unknown. Two mitochondrial proteins known to be involved in sensorineural hearing loss, CLPP protease, and Timm8a translocate, were chosen as the focus of this study. To investigate the role of these proteins in hair cell function and susceptibility, I used the zebrafish lateral line as a model. Lateral line hair cells are structurally and functionally similar to those in the mammalian inner ear, but instead detect changes in water flow. CRIPSPR/Cas9 was used to introduce dsDNA breaks in CLPP and Timm8a in zebrafish embryos. Non-homologous end repair following dsDNA breaks is error prone, and can introduce small insertions or deletions (indels) in the protein coding sequence which often leads to a loss of function in the protein. In order to quantify hair cell susceptibility, I treated embryos 5 days post-fertilization with varying concentrations of neomycin, a well characterized ototoxin, and counted the hair cells remaining after treatment. Future characterization of these mutants will involve examining mitochondrial activity and oxidation levels. These studies will provide insight into how mutations in CLPP and Timm8a affect hair cell function and susceptibility to damage. Additionally, they will more broadly inform our understanding of how changes in mitochondrial function can cause deafness.

Development of antibody-based diagnostics, biomolecular sensors, and biomaterial with reactive surface requires the ability to control the adsorption of bioactive protein at a surface. All medical devices exposed in an in vivo biological environment are immediately coated with a layer of adsorbed protein that plays an important role in the body’s behaviors towards the devices and the effectiveness of the devices themselves. In this study, we use a multi-technique approach to characterize the effect of protein G B1 (6 kDa) orientation on multi-layer antibody binding. By conjugating a cysteine group at different positions along the protein’s surface, we can immobilize five variants of the protein (V21C, D35C, E42C, T11C, and WT) onto maleimide and bare gold surfaces. We have confirmed through X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry experiments that the proteins adsorb onto these surfaces. In addition, we introduced primary IgG and secondary F(ab’)2 antibodies to investigate if the protein’s orientation will affect the binding of the antibodies. The adsorbed mass are measured using quartz crystal microbalance with dissipation monitoring. We have demonstrated that the cysteine mutants orient better on maleimide surfaces and encourage more IgG binding. We will also study the effect of temperature on adsorption mass and extend the experiments to amine and carboxyl functionalized surfaces. Fully developed techniques to accurately characterize the orientation and quantify the amount of protein and secondary adsorption will significantly contribute to understanding the interactions of biomaterials with the biological environment.

The embryonic development of vertebrates involves the anterior movement of progenitor cells, which form and proliferate at the posterior end of the embryonic body and then move anteriorly to differentiate into neural and muscle cells. Our interest is in understanding how these progenitor cells migrate, and how their movement is regulated, using the zebrafish as a model system because of its outstanding optical qualities. In my project, I am examining the role of specific proteins that are hypothesized to be important in this migration. To alter the expression of these proteins, I injected messenger RNAs encoding these proteins and also some newly developed transgenic lines where the expression of these proteins is regulated by a short period heat shock. For the mRNA injections I examined the phenotype of the whole embryo, whereas with the transgenic lines I microdissected the most posterior end of the growing embryo (called the “tailbud”) and imaged cell movement using a confocal microscope, with image analysis using software such as Slidebook and ImageJ. My results show that a change in the expression of several interesting proteins produced abnormal embryos and yielded abnormal movement of progenitor cells and impaired formation of the tail. These results provide useful information that helps us understand vertebrate development, including human embryogenesis.

During brain surgery for intractable epilepsy, clinicians utilize electrodes placed on the cerebral cortex to monitor seizure activity and map functional brain areas in patients so that the epileptic brain regions may be removed. This recording method is called electrocorticography (ECoG) and is essential to prevent the accidental removal of brain regions that are crucial for daily activities. To reduce tremors produced by Parkinson’s disease, electrical stimulation is applied to electrodes implanted within the deep brain near the subthalamalic nucleus. In both cases, to optimize treatment, a more detailed study of current flow during electrical stimulation than is currently done, due to surgical limitations of electrode placement, is needed. In addition, the time available to work with patients during brain surgery is extremely limited, necessitating a non-clinical research platform. I developed a saline-enriched gelatin phantom brain that mimics the electrical resistivity of the brain to study brain stimulation outside the surgery room, and I compared measurements taken with this model to analogous measurements of the brains of human patients made by my group. I found that my measurements agree within 10-15% with those made by my group on human brains. Using the phantom brain will allow us to easily and safely study electrical brain stimulation. Also, an electrical phantom that can be used offline to determine the most effective measurements will be extremely valuable to inform future research questions while working with patients. We are utilizing the phantom brain model to study how various electrode geometries and the cerebrospinal fluid layer that covers the brain have an effect on the amount of current that flows into the brain. Findings in our phantom brain will help us more effectively treat neurological disorders like intractable epilepsy and Parkinson’s and perform safer stimulations through optimized stimulation protocols.

The Cretaceous-Paleogene (K-Pg) mass extinction is the most recent of the ‘big five’ mass extinctions and led to the extinction of all dinosaurs except birds. Theropod dinosaurs, which were mostly carnivorous, were important members of latest Cretaceous terrestrial ecosystems. A popular hypothesis for the K-Pg mass extinction is that a meteor impact caused rapid extinction of many taxa 66 million years ago (Ma). The fossil record of dinosaur skeletons is not sufficient to assess the timing and pattern of the K-Pg mass extinction and its potential cause(s). Here, I documented theropod diversity using a sample of 783 fossil teeth from eight localities that span most of the depositional duration of the Hell Creek Formation (HCF; ca. 1.9 million years). To test the hypothesis that the bolide-impact was the sole cause of the mass extinction, I wanted to determine if significant changes in theropod diversity occurred before the impact at the K-Pg boundary. New and updated taxonomic identifications were based on the current literature. Raw, range-thru, and rarefied taxonomic richnesses as well as relative abundances and heterogeneity indices (evenness, Simpson’s) were calculated for each of the fossil localities. In total, eight theropod taxa were identified from the HCF. Richness shows a slight decrease from eight taxa in the lowermost HCF locality to five to seven taxa higher in section. Rarefied richness was lowest in the uppermost HCF. Although the changes observed in richness are minor, relative abundance and heterogeneity values differed significantly from the middle to upper HCF localities implying that major changes in theropod faunas occurred locally within the last few hundred thousand years of the Cretaceous prior to the bolide impact. Similar patterns have been observed in the amphibian and mammalian faunas suggesting other environmental factors (e.g., volcanism, climate change) contributed to the causes of the mass extinction.

The loss of mechanosensory hair cells in the inner ear gives rise to many hearing and balance-related disorders. The most common causes of acquired hair cell loss are normal aging, ototoxic medications and frequent exposure to unusually loud environments, noise exposure. Two main drug classes, aminoglycoside antibiotics and cisplatin-based chemotherapeutic drugs, are known to be ototoxic, with usage resulting in varying levels of temporary or permanent hearing loss through hair cell death. Our goals are to understand and eventually prevent hearing loss due to treatments with these drugs. Because of the inaccessibility and complexity of the mammalian inner ear, hair cells in the zebrafish lateral line have emerged as an outstanding system for studying hair cell death. The external location of the hair cells allows for direct access and visualization of the hair cells, in order to study the cellular pathways underlying hair cell death, and for large drug library screens to identify compounds that protect against ototoxic effects. As a result of a drug screen and subsequent compound analog analysis, a compound named ORC-13661 has been found to be robustly protective against several aminoglycosides in a dose-dependent manner. We have found that there are distinct mechanisms that underlie antibiotic-induced toxicity, distinguished by different temporal events. Some aminoglycosides, including neomycin, kill hair cells rapidly, within one hour of exposure. Other aminoglycosides, including gentamicin, have a much slower time course, taking up to 24 hours to kill hair cells after an initial one-hour exposure. We find that ORC-13661 provides maximal protection to both classes of aminoglycosides if it is present at the same time as the toxic compound. Additionally, the delayed aminoglycoside exposure paradigm gives us an opportunity to use compounds with known effects on cell death pathways to shed light on new mechanisms underlying hair cell death.

Atherosclerosis is a disease in which cholesterol accumulates in the walls of arteries, limits blood flow, and causes heart attacks and strokes. We aim to prevent and reverse atherosclerosis with a gene therapy that increases levels of apolipoprotein A-I (apoA-I), a protein that removes cholesterol from the artery wall. This therapy increases expression of apoA-I specifically in endothelial cells (EC), cells that line the artery wall. Our initial vectors for expressing apoA-I contained the murine endothelin-1 (mET-1) promoter (an EC-specific promoter) both without (1XETE) and with (4XETE) three additional copies of an endothelin-1 enhancer (ETE). We used two approaches to increase apoA-I expression: 1) literature searches for regulatory sequences that increase EC gene expression in other contexts; 2) novel bioinformatics approaches that identify EC-specific cis-regulatory sequences (CRMs) throughout the genome based on ENCODE data, relative local abundance of EC-related transcription factor binding sites, and interspecies conservation. We ligate regulatory sequences into the apoA-I vectors mentioned above. We transfect these constructs into cultured EC and measure apoA-I expression. Using the first approach, we found that addition of a shear-stress-response element into the 4XETE vector did not alter apoA-I expression. Addition of a single MEF2C enhancer sequence into the 4XETE vector decreased expression; however, after this decline, addition of more copies of the MEF2C enhancer increased apoA-I expression. Lower expression may be due to unfavorable interactions between multiple enhancer elements. Using the second approach, we identified eleven CRMs (55-352 bp) clustered around the promoters of genes highly expressed in EC: CDH5, EFEMP1, THBS1, and VWF. In conclusion, trial-and-error addition of enhancers has not reliably improved EC apoA-I expression. A bioinformatics-based approach identified CRMs associated with high transcriptional activity in EC. We inserted the eleven CRMs into both the 1XETE and 4XETE apoA-I vectors and will test whether they increase EC apoA-I expression.

The goal of this study was to understand more precisely the placement of the Scalesia clade within Helianthinaea– specifically what taxa are sister to Scalesia and what traits diverged between Scalesia and its sister clades. Previous research has shown Scalesia to be monophyletic and placed Scalesia as sister to Pappabolus and basal to Viguiera, but with low support due to homoplasy. We used extracted ITS, ETS, and psbA sequence DNA from seventeen Scalesia samples from the Galapagos and sequence DNA from ninety two species from mainland South America, North America, and Mesoamerica that we obtained from Genbank. Bayesian analyses of concatenated ITS +ETS + psbA sequence data, maximum likelihood analyses of ITS+ETS+psbA/ITS+ETS, and individual gene region data implied that Scalesia is sister to Pappabolus, Heiseria pusilla, Viguiera simsioides, and Syncretocarpus sericeus with high likelihood support and is monophyletic consistently throughout all trees. Our data also implied that all Scalesia taxa is very closely related and very recently diverged. An interesting split between a woody ancestor (Zaluzania grayana) and an herbaceous, derived species (Scalesia baurii) was observed. Further taxonomic work, especially on the reclassification of Aldama and Viguiera is still necessary.