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Investigating the Interaction between Pseudophosphatase MK-STYX and Vimentin
No full textMK-STYX [MAPK (mitogen-activated protein kinase) phosphoserine/threonine/tyrosine-binding protein) is a pseudophosphatase belonging to the MAPK phosphatase (MKP) subfamily within the protein tyrosine phosphatase (PTP) superfamily. The absence of a histidine and critical cysteine in the signature active motif render MK-STYX catalytically inactive, earning it the prefix pseudo. A mass spectrometry study revealed that MK-STYX and the cytoskeletal protein vimentin are binding partners. Vimentin is expressed in the early stage of neuronal development, contributes to organelle organization, and localizes at the perinucleus, mitochondria, and golgi. It has been reported that the downregulation of vimentin decreases neuritogenesis. Previous research revealed that MK-STYX increases neurite outgrowth in rat pheochromocytoma PC-12 cells and rat hippocampal primary neurons. This interaction of MK-STYX and vimentin and the role of MK-STYX in neurite formation point towards a possible relationship between the two proteins that may contribute to neuritogenesis. Our goal is to further confirm and characterize the interaction between MK-STYX and vimentin. HEK-293 cells were cotransfected with the following mammalian expression vectors: GFP and mCherry, GFP-MK-STYX and mCherry, GFP-MK-STYX(active mutant) and mCherry, mCherry-vimentin and GFP, GFP-MK-STYX and mCherry-vimentin, or GFP-MK-STYX(active mutant) and mCherry-vimentin followed by western blot analysis. Vimentin expression was increased in the presence of MK-STYX. To further characterize the dynamics between MK-STYX and vimentin, we performed colocalization experiments and used fluorescence microscopy to analyze them. The conditions were GFP, mCherry, GFP and mCherry, GFP-MK-STYX, mCherry-vimentin, GFP and mCherry-vimentin, GFP-MK-STYX(active mutant) and mCherry-vimentin, or GFP-MK-STYX and mCherry-vimentin. Higher rates of partial colocalization were observed in GFP-MK-STYX and mCherry-vimentin conditions (≥75%) than in the mCherry-vimentin and GFP condition, confirming the pattern observed in our pilot colocalization experiments. All instances of colocalization were observed in the cytoplasm and future studies will investigate vimentin localization at the perinucleus, mitochondria, or golgi in the presence of MK-STYX and GFP-MK-STYX(active mutant) which has phosphatase activity. Taken together the colocalization pattern of MK-STYX and vimentin and the increase of vimentin in the presence of MK-STYX support the hypothesis that MK-STYX and vimentin are interacting and together may serve as important players in neuritogenesis.BiologyBachelors of Science (BS
“I Maintained My Authority:” Gender and Power in the Eighteenth-Century Virginia Home
No full textThis thesis examines William Byrd II and Lucy Parke Byrd alongside John Custis IV and Frances Parke Custis as significant players in the early Virginia landscape. I base the bulk of my work on William Byrd II’s diaries, John Custis IV and Frances Parke Custis’s Marriage Agreement, and the homes of these two men. Where I do not have Lucy and Frances’s voices, I am reliant on the words of their husbands – what they did say, and at times, what they omitted. Without a clear image of the Byrds’ and Custis’ world, Lucy and Frances might appear an insignificant, but unwelcome burden to these men. The object of William Byrd and John Custis’s world was to “maintain authority;” when we reconsider what it means to succeed in that effort and examine the impact these men’s wives had on their success as patriarchs, it becomes clear that these men were only nominally successful. As the authors of surviving written documents, they get the last word, but the voices of women like Lucy Parke Byrd and Frances Parke Custis are still incredibly clear within them.HistoryBachelors of Arts (BA
HerbertWM and Beyond: A Multi-Faceted Approach to Understanding and Engineering Bacteriophage Host Range
No full textIn order for bacteriophage to be useable as reliable and efficient tools in synthetic biology, real-world problems, and other scientific applications, there must be a way to screen efficiently for viruses when targeting specific hosts, especially on the strain-level; in the real world, bacteria constantly mutate and evolve, developing anti-phage defense mechanisms and diverging significantly from model strains used in the lab. Additionally, there is a dire need for further characterization and engineering of phage and phage-satellite circuits in lesser-studied hosts. This thesis contributes to the field of bacteriophage research through a multi-faceted approach to host range prediction and phage engineering. By manually compiling a comprehensive dataset of all publicly available E. coli and Mycobacteria phage host range assays, this work presents the first infectivity prediction model to incorporate confirmed incompatibility features alongside compatibility and homology metrics. The E. coli model demonstrates consistent strain-level accuracy, addressing the high-resolution specificity required for targeting dynamic bacterial populations beyond static lab strains. Preliminary engineering efforts using the pYS2/pYS1 system lay the groundwork for modifying phage HerbertWM, offering a promising strategy to refine its host range and develop a controllable lysogenic switch for targeting Mycobacteria. Lastly, the characterization and initial engineering of phagelet genomes provide both practical tools and foundational insights into these understudied genetic elements, opening new avenues for synthetic biology applications.BiologyBachelors of Science (BS
Unearthing the Historiography of Richard III: British Identity, Historical Memory, and Legacy Across Five Centuries
No full textRichard III, King of England from June 26th, 1483, to August 22nd, 1485 encapsulates a fascinating mystery for many people. Why does such a figure sustain personal connections and produce strong emotions across time? This thesis traces the historiography and popular memory of Richard III across five centuries and asserts that changing understandings of British identity – national, local, and personal – from the seventeenth through the twenty-first century reflect varied depictions of him. The historiography of Richard indicates that figures of the more ancient, imagined past – not just the re-examination and re-evaluation of more recent historical figures – demonstrate the impact of such evolving views on how Britons today define themselves. Richard’s legacy has been debated for centuries in Great Britain, sparking interest among historians and non-historians, such as archaeologists, authors, courtiers, statesmen, and theologians. Much of Ricardian historiography, pro- and anti-, is interlaced with literary fiction and theatrical drama, complicating the scholarship. Thus, Richard III provides an excellent case study for comprehending how scholars of varied fields and non-scholars can unintentionally impact written history and how contested and changing identities across generations influence historiography.HistoryBachelors of Arts (BA
Exploring MK-STYX's Interaction with KDAC6 and Dynein
No full textThe pseudophosphatase MK-STYX (mitogen activated protein kinase phosphoserine/threonine/tyrosine binding protein) is a member of the MAPK phosphatase (MKP) family. However, MK-STYX has the amino acid sequence FSX5R in its active site instead of the sequence HCX5R present in other protein tyrosine phosphatases. The replacement of the critical cysteine residue with a serine renders MK-STYX catalytically inactive as a phosphatase. The Hinton Lab previously reported that MK-STYX interacts with the RNA binding protein Ras-GTPase SH3 domain binding protein-1 (G3BP1) in the stress response pathway. During the stress response, mRNA translation is halted, and stress granules, which are cytoplasmic aggregates of mRNA and mRNA binding proteins, are formed. G3BP1 serves as a major structural component of these stress granules. Additionally, microtubules and their associated proteins transport stress granule components to the correct places during the initial stages of assembly. Many proteins that move along microtubules to stress granules, including G3BP1, require the activity of two proteins: dynein and lysine deacetylase 6 (KDAC6). KDAC6 is responsible for facilitating the transport of ubiquitinylated proteins by dynein. Additionally, it regulates microtubule reorganization through the deacetylation of polymerized microtubules. Dynein motility and localization can be affected by several proteins and alteration of these two dynamics may affect stress granule formation. MK-STYX decreases stress granule assembly, however, it does so independently of G3BP1 phosphorylation status at serine 149 which has been reported to induce stress granule formation. Given the essential nature of G3BP1’s interaction with KDAC6 and dynein, we decided to investigate what effects if any that MK-STYX is having on the two proteins. Because KDAC6 deacetylase activity has been shown to affect stress granule assembly, we investigated whether MK-STYX influenced KDAC6 deacetylation. KDAC6 activity assay pilot studies indicate that KDAC6 deacetylase activity is unaffected by the presence of MK-STYX. To further understand the interaction between MK-STYX and dynein, we transfected HEK-293 cells with fluorophore conjugated constructs and studied their localization. Dynein appears to colocalize with MK-STYX. Additionally, dynein localizes to aggregates in the presence of MK-STYX. We have previously demonstrated that MK-STYX does not localize to G3BP1-induced stress granules, so these dynein aggregates may be a novel way of controlling dynein activity.BiologyBachelors of Science (BS
The Realities of Rape: Understanding the Foundations of a Woman’s Space in Ancient Greece and Rome
No full textTW: rape, sexual violence, abortion and trauma.Sexual violence has been a subject feared by scholars for quite some time, even more so in ancient cultures of Greece and Rome. This thesis used modern trauma and rape theory, medical texts, and legal codes from antiquity to explore the ramifications rape had on real ancient women. The way in which a woman's space in a patriarchal society was formed is reflected largely on how her rape was handled. From abortive methods to the legality behind consequences for a rapist, it is revealed how little support ancient women had despite there being constant conversation surrounding the need for her status to be unviolated to fulfill her societal roles.Classical StudiesBachelors of Arts (BA
Characterizing a Novel Phage Particle: The Phagelet
No full textPhages are viruses that infect bacteria. They are the most abundant particle in our biosphere. Mycobacteriophages that infect mycobacteria have important applications for human health and genetic engineering. This research describes the discovery of a novel phage particle called a phagelet that is similar to mycobacteriophages but lacks tail proteins and a lysin cassette. This phagelet infects Mycobacteria aichiense which contains the prophage HerbertWM. The presence of an additional phage in this system further makes this system unique and suggests that perhaps the phagelets are borrowing proteins from the existing prophage. I discuss the characteristics of a phagelet as well as discuss tools to analyze the function of these phaglets including the development of an RNA-Seq protocol and the creation of Golden Gate Fragments. This research contributes to increasing our understanding of mycobacteriophage genetics.BiologyBachelors of Science (BS
Emotional Competence in Children with Incarcerated Parents: Evaluating the Impact of the Milk and Cookies (MAC) Program
No full textWhile existing literature has emphasized the importance of social-emotional learning (SEL) in early childhood development, few interventions have been tailored specifically to the unique needs of children experiencing parental incarceration. The present study evaluates the Milk and Cookies (MAC) Program, a trauma-informed, school-based SEL intervention designed for elementary-aged children with an incarcerated parent. Using a quasi-experimental design, this study assessed emotion identification accuracy and anger bias among 34 kindergarten and first-grade participants across an academic year. Emotional competence was measured using the facial expression section of the Assessment of Children’s Emotion Skills (ACES) at two time points. Results indicated no significant differences in emotion identification or anger bias between children who had completed a prior year of MAC programming and those who had not. Similarly, no significant within-year changes were observed in emotion identification, though gender differences were noted, with girls displaying significantly higher emotion accuracy scores than boys at baseline (p = .019). Despite a lack of statistically significant program effects, slight reductions in anger bias suggest potential subtle gains over time. These findings underscore the importance of extended and contextually sensitive interventions for high-risk populations. Future research should incorporate larger samples, comparison groups, and multi-informant measures to more fully capture the impact of SEL interventions on children with incarcerated parents. Keywords: parental incarceration, social-emotional learning, emotion identification, anger biasPsychologyBachelors of Science (BS
The Unseen Vaccine: Trans-Imperial Networks, Medical Exploitation, and State Formation in Nineteenth-Century South America (1795-1825)
No full textFollowing Edward Jenner’s vaccine discovery, leaders entered the nineteenth century with the renewed hope of curbing the smallpox epidemic. In 1803, Spain’s Real Expedición Filantrópica de la Vacuna (Royal Philanthropic Vaccine Expedition) set sail to disseminate the smallpox vaccine throughout Spanish America and Asia. Scholarship on vaccination in nineteenth-century Latin America has focused on this expedition; however, historians have neglected peripheral regions untouched by the mission. The Viceroyalty of the Río de la Plata was one such area. This thesis examines how the inhabitants of the Viceroyalty of the Río de la Plata obtained a vaccine for smallpox, despite the Spanish Empire’s failure to ensure the vaccine's arrival in the region. I argue that Río de la Plata’s trans-imperial commercial networks and pervasive local agency, anchored in the regional fusion of Catholic-Enlightenment ideas, made possible the successful dissemination of the smallpox vaccine in the Viceroyalty of Río de la Plata and was instrumental in the creation of the newly independent state in the region after 1810. While unofficial expeditions were active in the Caribbean and Brazil, the vaccine’s pathway in the Río de la Plata was uniquely commercial, with trade precipitating the vaccine’s arrival and mercantile interests driving vaccination campaigns. Commercial, administrative, and religious elites centered these mercantile-medical ventures within a Catholic-Enlightenment ideology, through which they used the language of humanitarianism to justify profitable vaccine enterprises and the medical exploitation of enslaved people as commodified vaccine carriers. These elites’ autonomy persisted into the revolutionary period, where elites’ standardization of vaccination and distribution procedures, in which administrative actors and individual elites disseminated the vaccine, legitimated the revolution and contributed to processes of state formation.HistoryBachelors of Arts (BA
Cascade Aza-Prins Reactions to Access Novel Fused Tetracyclic Heterocycles
No full textHerein, we explore the scope and limitations of the BF3•Et2O-mediated alkyne aza-Prins reaction in tandem with an intramolecular Friedel-Crafts addition to synthesize tetracyclic molecules with both nitrogen and oxygen-containing heterocycles. We demonstrate its utility and versatility with the synthesis of numerous compounds containing a 7,8-dihydro-6H-chromeno[3,4-c]quinoline core. The use of aldehydes with varying electronic properties, ranging from both electron-donating to strongly electron-withdrawing, as well as varying substituents on the alkynyl substrates all afforded our desired target molecule in modest to good yields.ChemistryBachelors of Science (BS