Clinicians in low-resource settings face several challenges when using nucleic acid amplification to diagnose disease at the point of care, because conventional methods have high costs per sample, require trained technicians, and require the use of expensive equipment which relies on grid-supplied electricity. The first step of diagnosis using nucleic acid amplification is release of amplifiable DNA from the cell. The AudioLyse is an inexpensive lysis device, which is powered by any audio device (such as a cell phone or any small mp3 player, both of which only require charging before reuse). Lysis takes just 10 minutes in a “set it and forget it” format. I show that the AudioLyse is capable of lysing methicillin-sensitive Staphylococcus aureus (MSSA). I compared the lysis of MSSA using the AudioLyse method, to lysis with the Bead Beater, a standard lysis technique. I found the AudioLyse effectively released amplifiable DNA from MSSA cells. I also tested the effects of glycerol and simulated nasal matrix, or SNM, on the lysis method’s ability to release amplifiable DNA. I found that high levels of simulated nasal matrix decreased the efficiency of amplifiable DNA release when both the bead beater and AudioLyse were used. However, we expect nasal swabs will be AudioLyse compatible, after dilution in lysis buffer. I found that the AudioLyse doesn’t perform well in releasing amplifiable DNA in viscous solutions. Glycerol was added to samples to test how the AudioLyse’s efficiency was affected. The AudioLyse displayed a statistically significant decrease in amplifiable DNA released in a 50% glycerol solution, releasing just 7.8% of the amplifiable DNA the bead beater was able to release. As of now, the AudioLyse is an efficient device in terms of releasing sufficient amplifiable DNA for disease detection, but remains limited by the viscosity of samples. Future work will include testing the AudioLyse on other genera of bacteria.

The rise of multidrug resistant bacteria necessitates finding alternative therapies against bacterial pathogens. Virulence in many pathogenic bacteria are predicated on their ability to communicate and act in unison. In a phenomenon known as quorum sensing (QS), a threshold mass of bacterial cells produces an amplified chemical signal that triggers the collective expression of a new phenotype, such as enhanced virulence. Therefore, inhibiting QS, without preventing cellular growth, may be a recourse against drug resistant bacteria. Marine invertebrates found in warmer waters, such as the Australian reef and Indian Ocean, have shown quorum sensing inhibitory (QSI) potential. However, most marine invertebrates in the Pacific Northwest’s Puget Sound have not previously been screened for QSI compounds. This research presents the findings for QSI sources among Puget Sound’s marine sponges and tunicates, using a bacterial reporter strain to screen for QSI activity. We used the reporter strain Chromobacterium violaceum (ATCC 12472), which uses the QS signal n-Acyl-homoserine lactone (AHL) to produce its purple pigment. QSI activity was screened for by overlaying the reporter strain with a sponge or tunicate sample. Subsequent absence of pigment in the indicator strain suggested the presence of a QSI compound. Promising compounds were then extracted and identified using analytical techniques. These results help expand the catalogue of known QSI molecules from marine sources, advancing potential therapies against existing and future antibiotic resistant pathogens.

In the humanities, histories of technology generally examine objects through representations, such as patents, illustrations, fictions, and newspapers. With objects flattened on the page, this approach rarely attends to how technologies worked in context and over time. In response, we prototyped a series of early technologies, which cannot be immediately accessed in museums or archives; they have been lost, destroyed, or broken, yet they deeply inform present day devices. In this talk, we present three of our prototypes: an early wearable technology, an early wire recorder, and an early reading machine. The early wearable prototype features a skull cravat-pin (designed by Gustave Trouvé and Auguste-Germain Cadet Picard in 1867), which was said to snap its jaw and roll its eyes. We replicated the mechanism of the skull using electromagnetic technologies available in the nineteenth century. We found that the size of the skull, as described in historical records, could not have contained the electromagnet necessary to propel the skull’s jaw. This suggests that textual representations of the stick-pin were distorted by Victorian hyperbole. Elsewhere, our wire recorder prototype recreates a precursor to data storage: Valdemar Poulsen’s 1898 experiments with sound, whereby people yelled into a transmitter while running a trolley along a wire. Despite its claims for high fidelity, our findings suggest this process was highly contingent on the bodies and movements of users. Finally, the reading machine prototype explores the maintenance of Fournier d’Albe’s 1913 optophone, which translated images into sound, by one of its blind users, Mary Jameson. Early results of this research stress that Jameson contributed significantly to the optophone’s development, even if she is never mentioned in patents and rarely in other textual accounts. Together, these prototypes highlight why trial-and-error experimentation (as a method for humanities research) matters for historical inquiry into technology off the page.

Fostering students’ motivation and interest in higher education composition classes is vital to sustain student engagement. However, short-term motivation does not necessarily determine long-term success; recent research has found that an individual’s level of grit plays a significant role in his or her long-term success (Duckworth, Peterson, Matthews, & Kelly, 2007). To investigate the role of grit in writing classrooms, we are exploring how writing instructors can foster students’ grit for a greater chance of long-term success. Because Duckworth et al. (2007) consider interest and effort to be crucial factors in grit, we chose to examine whether or not service-learning (SL) can foster students’ grit, as SL been shown to increase student engagement in the past. Using Duckworth et al.’s eight-question grit scale, this project investigated the impact of SL on the grit of students in basic writing classes at Shippensburg University in Pennsylvania, comparing three SL classes (Group 1) and two non-SL classes (Group 2). Data was analyzed using t-tests to compare the average grit scores between the groups at mid- and post-semester. Although results indicated no significant correlation between grit and SL, this study suggests that grit stems more from internal rather than external motivators. Thus, this research can potentially open up a conversation about factors and influences that affect an individual’s grit.

Considering the current price and competitiveness of higher education, extra-institutional programs like Advanced Placement, International Baccalaureate, and Running Start are playing an increasingly critical role in public education. Unfortunately these programs, varying in curriculum, course structure, assessment, and aptitude, are not always consistent with the learning outcomes of universities. At University of Washington, Texts and Teachers, a part of the UW in the High School Program, attempts to bridge the gap between extra-institutional college curricula and university learning outcomes by creating collaborative course design, teaching common texts, using common culminating writing portfolios for assessment, and scheduling reciprocal faculty visits to high school classrooms and high school class visits to the linked UW classrooom. In order to assess the program’s success in meeting common learning outcomes, I have designed a research project guided by program instructor Professor Gary Handwerk which evaluates student portfolios for their demonstration of writing ability relative to the UW Expository Writing Program’s learning outcomes. Using a text-response oriented scoring pathway taken from Linda Flower’s 1990 text Reading to Write, I reviewed student portfolios from ENGL 111 and from T&T classes both on and off campus, assessing three primary trait categories: organizational strategies, concept development, and critical analysis. Once scored, I tested for statistically significant differences between class setting and course level and concluded that there was no significant difference between classes. In addition, this methodology distinguished average rates at which students improve their writing ability in each setting. This research provides evidence to support the compatibility of T&T and EWP writing instruction. It thus demonstrates the success of extra-institutional college curricula operating in a context specific to the learning outcomes of a particular university.

In order to successfully invade and survive within a host cell, pathogens such as the bacteria Shigella flexneri have to suppress the host cell’s immune response. To do this, Shigella flexneri inject bacterial effector proteins into the host cell that hamper the host cell’s ability to detect and respond to invasion. One such protein is IpaH4.5, a bacterial E3 ubiquitin ligase, which hijacks the host cell’s ubiquitylation machinery to transfer ubiquitin, a small signaling protein, to a substrate. Ubiquitin signaling impacts all aspects of eukaryotic cell biology. Often modification of a protein by ubiquitin serves to mark protein for destruction. IpaH4.5 targets proteins involved in the host cell’s immune response, dampening the host cell’s ability to respond to invasion by Shigella flexneri. I am investigating IpaH4.5 and its interactions with both host substrates and the host ubiquitin machinery. After sub cloning out the various domains of TbK1, a known substrate of IpaH4.5, I will perform ubiquitilation assays in order to determine which domain of TBK1 is polyubiquitilated, indicating that IpaH4.5 interacts with this domain. Then, this domain can be used for structural and mechanical analysis of IpaH4.5, such as determining how mutations in IpaH4.5 affect ubiquitilation through biochemical assays. Understanding how IpaH4.5 binds to and interacts with its substrates will increase our understanding of how Shigella flexneri dampens host immune response as well as help us better understand the mechanisms of bacterial E3 ubiquitin ligases. Since IpaH4.5 functions differently than eukaryotic E3 ubiquitin ligases, increasing our understanding of its mechanisms could allow us to use it as a target for future drug development. Finding new targets for drug development is becoming increasingly important with the spread of antibiotic resistant bacteria.

Writing is both emotional and intellectual task. For writing tutors, it is not rare to encounter writers asking for affirmation on their work, regardless of the quality of works and confidence level of writers. There is a bias, or expectation, that one’s role as a tutor is to provide intellectual support on the writing material. Thus, there is little literature or discussion on how tutors may provide additional types of support to the writer. Recognition of emotional support in the writing center is significant as it creates a perspective shift on the role of writing tutors and the function of writing centers. The research is designed to observe how often writers seek emotional support (e.g. demand affirmation or approval) from tutors in addition to intellectual support (e.g. grammar checks) and how current tutors engage in providing emotional support. Data for the study is drawn from 40 both undergraduate and graduate tutors who are currently working actively at Odegaard Writing and Research Center. The tutors participated in an anonymous self assessment survey online in which they were asked to identify how frequently they feel, encounter, or engage in specific behavior. The questionnaire is divided into three parts: Tutors’ emotional labor, Tutors’ identification of Writers’ demanding of emotional support, and Tutors’ accommodation of Writers’ emotional demand. This study found 72% of tutors encounter a writer’s emotional expression, 69% of tutors spend time to build a personal connection with a writer, and 67% of tutors often to always try to comfort stressed writers. The findings indicate consistent demand in emotional support from writers and emotional labor on part of tutors. This provides a foundation for further research to be conducted in exploring the roles of academic tutors beyond providing intellectual support.
 

The oxygen evolving complex (OEC) is the enzyme in Photosystem II responsible for oxidizing water into oxygen gas. The OEC, a Mn₄O₅Ca cofactor, has one manganese “dangling” off a cubane, and it undergoes several conformational changes as photons provide energy for the splitting of water. The mechanisms and intermediates for the transitions between states are not fully understood, so research into manganese dioxygen chemistry will illuminate this important biological process. In order to characterize intermediates more successfully, we use synthetic, small molecule, ligand framed analogues that emulate the properties of the enzyme. Synthetic analogues have a distinct advantage in characterizing intermediates because the enzyme is limited to activity under conditions of a living organism such as temperature and solvent. By tuning the conditions and coordination sphere of our manganese complexes, we can stabilize metastable intermediates in order to spectroscopically characterize them. Our target is to synthesize a mononuclear manganese superoxo complex modeling the dangling manganese, which has been previously reported as an active site in oxygen evolution. A superoxo must be the last intermediate in oxygen evolution, so by characterizing our complex, we can determine viable mechanisms for the evolution of oxygen off the dangling manganese. We designed the ligand to be sterically bulky and have a low coordination number so there is a selective open coordination site for dioxygen binding. The lower coordination number raises the manganese’s Lewis acidity which allows for reversible oxygen binding. A trans thiolate is incorporated into the ligand framework as it has been previously shown that the sulfur to metal charge transfer band is in the visible region, giving the complex an easily identifiable spectroscopic property. By synthesizing our superoxo complex then characterizing its properties, we can better understand the complex mechanisms of oxygen evolution.