Conditional gene expression systems that enable inducible and reversible transcriptional control are essential research tools and have broad applications in biomedicine and biotechnology. The reverse tetracycline transcriptional activator is a canonical system for engineered gene expression control that enables graded and gratuitous modulation of target gene transcription in eukaryotes from yeast to human cell lines and transgenic animals. However, the system has a tendency to activate transcription even in the absence of tetracycline and this leaky target gene expression impedes its use. Here, we identify single amino acid substitutions that greatly enhance the dynamic range of the system by reducing leaky transcription to undetectable levels while retaining high expression capacity in the presence of inducer. Furthermore, we show that these improved DNA binding domains can be fused to repression domains to create synthetic transcriptional repressors. The function of these transcriptional repressors is dependent on the location of their recruitment and their mechanisms of action.

Indiana University-Purdue University Indianapolis (IUPUI) / Chronic pain is a health problem that impacts at least 10% of the world population. Social support has beneficial effects for those who have chronic pain. Social support can be emotional, where a supporter is present, validating, and assuring; or instrumental, where a supporter provides services to address an imbalance in needs. Despite the benefits of social support, relatively little attention has been given to willingness to provide support to people with pain during transition to adulthood. The current study explored gender differences and the role of attitudes towards emotional expression in young adults’ willingness to provide emotional and instrumental support to a virtual human peer with pain. Participants (N=234, mean age=20) viewed videos/vignettes of a male or female peer with pain and responded to questions about willingness to provide support and attitudes towards emotional expression. Female participants were more willing to provide support than male participants, F(1,218) = 27.37, p < 0.01, and participants were more willing to provide emotional support compared to instrumental support, F(1,218) = 6.72, p = 0.01. Attitudes towards emotional expression mediated the relationship between participant gender and emotional and instrumental support (total indirect effect = 0.05; 95% CI [.01-.12]; total indirect effect = 0.05; 95% CI [.01-.12]). There was not an interaction between participant and peer gender for emotional, P = 0.26, or instrumental, P = 0.47, support. The results suggest that women are more willing to provide emotional and instrumental support and that fewer negative attitudes about emotional expression help explain the relationship. The findings also suggest that the gender of the peer does not impact willingness to provide support.

This project examines the changing shape of femininity and masculinity for Marines from World War I to the Korean War, focusing on the ways that the body serves as a canvas for demonstrating the negotiation of gender roles and the Marine Corps image. Gender has been a constant issue for the military. However, few historical studies consider the ways that the Marine Corps’ status as a particularly elite, masculine institution impacted the desired image of femininity for its female recruits and how this image changed over time. The hyper-masculine nature of the military influenced the relationship between masculinity and femininity for both servicemen and women. My project looks at these changes in masculinity and femininity by placing gender identity within the context of the hyper-masculine military environment. R.W. Connell’s Masculinities, Anthony Rotundo’s American Manhood, and Aaron Belkin’s Bring Me Men assist in putting gender identity in the military into a more complex and nuanced context, especially focusing on masculinity’s centrality to the American military institution. Belkin, in particular, argues that military masculinity has never been entirely devoid of feminine elements. Aspects of femininity have long been a part of military life, from domestic responsibilities often associated with women to close same sex companionship between soldiers. While generally considered less masculine when taken as separate behaviors, they did not seem problematic in a military context. This leads to the conclusion that the incorporation of women into the military was not a radical introduction of femininity into a solely masculine environment, but rather a more complicated shift in the relationship between gender and occupation. This project’s conclusions support this kind of closer relationship between masculinity and femininity in the military context. Francine D’Amico and Laurie Weinstein’s Gender Camouflage, Melissa Ming Foynes, Jillian C. Shipherd, and Ellen F. Harrington’s “Race and Gender Discrimination in the Marines,” Melissa S. Herbert’s Camouflage Isn’t Only for Combat, Heather J. Höpfl’s “Becoming a (Virile) Member: Women and the Military Body,” Leisa D. Meyer’s Creating GI Jane, and Sara L. Zeigler and Gregory G. Gunderson’s Moving Beyond GI Jane address this shift in gender relations and the resulting tension between military men and women throughout the twentieth and twenty-first centuries I investigate changes in military gender identity by looking at legislation and regulations controlling gender and sexuality in the military, media depictions of Marines, and the ways that gendered military identity plays out on the body, especially through physical fitness, uniforms, and bodily maintenance. The Marine Corps documented their ideas of normative masculine and feminine Marine bodies through pictures, propaganda, and newsletters. Examination of these different characteristics of the ideal body allow for comparison through time of the ways that Marines presented themselves to society, as well as the methods the Corps utilized to encourage images advantageous to its purposes. Such comparisons show changes in the perception of gender identity through time, as well as new norms of appearance and behavior that developed. This evidence illustrates the complicated and often contradictory relationship between masculinity and femininity that all Marines, male and female, negotiate. This project illustrates the significance of these frequently gendered representations of Marine bodies through time. They show the negotiation of gender within the Corps and how assumptions of gender roles shifted from one war to the next. Understanding these changes helps explain the tensions and conflicts which developed between male and female Marines during different periods, as well as creating a framework for investigating these tensions into the contemporary era. The primary sources used for this project focus on the appearance of Marines, male and female, and include national legislation related to Marines and military regulations enforcing conformity in dress and appearance. Memoirs of Marines, publications intended for Marine readers, as well as publications depicting Marines aid in gaining a better idea of the function of gender for Marines, especially in relation to their interactions between male and female Marines. These documents show the changes occurring in expectations about femininity and masculinity in the Marine Corps over time. Public publications, such as general interest magazines, women’s magazines, and newspapers, showed public ideas of Marines’ gender and their relationship to civilian American gender ideals. This project explores the changing shape of normative Marine Corps bodies and the impact of ideas of masculinity and femininity in their deployment as methods of supporting the services’ goals. / A Dissertation submitted to the Department of History in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Spring Semester 2019. / February 8, 2019. / Gender, Korean War, Marine Corps, US Military, World War I, World War II / Includes bibliographical references. / Suzanne Sinke, Professor Directing Dissertation; Dennis Moore, University Representative; Kurt Piehler, Committee Member; Charles Upchurch, Committee Member; Jennifer Koslow, Committee Member.

The thesis investigates how body movements influence a musical performance, with the aim to reach a more expressive performance by an increased awareness of gestures. In the study, three versions of the same clarinet piece were recorded on video; one with me, one with my clarinet teacher and one with a fellow clarinet student. The study addresses the following research questions: - How do body movements correspond to musical intentions? - How are my gestures formed and influenced by my teachers’ gestures? - In what ways can a raised awareness of gestures affect my musical performance? The videos were coded and analysed using open coding. As a reference, each clarinettist notated their intended phrasing in the score. This was marked as phrases (slurs) and Goal Points, and was also annotated in ELAN. With the intention to answer the first research question, the body movements were compared with the performers intended phrasing. In order to answer the second research question, coded sequences from each performance were compared with each other to find similarities and differences, using both quantitative and qualitative methods. Finally, I recorded a second performance of the same piece to investigate whether awareness of gestures affected my performance. Results align with previous research and indicate that body gestures are unique for each performer and connected to musical intentions. Results also indicate a resemblance in movement patterns between my teacher’s performance and my own, suggesting that gestures can be transferred from teacher to student.

The pallid sturgeon (Scaphirhynchus albus) is a federally endangered species, endemic to the Mississippi River drainage, stretching from the upper Missouri River in Montana to the Mississippi River, and continuing to the Gulf of Mexico. They are largely sympatric throughout this range with a close congener, the shovelnose sturgeon (S. platorhynchus), although speciation may have occurred when they were isolated in different refugia. In this study, we examined gene expression differences among pallid and shovelnose sturgeon families in response to heat stress. Gene expression can be considered a phenotype, and therefore, variability in expression can have an adaptive role in species. Additionally, we compared our results to a previous expression study that utilized RNA-Seq. We developed viable primer pairs for five genes in order to conduct RT-qPCR assays. There were significant differences in heat stress response between pallid and shovelnose sturgeon, potentially indicative of different evolved stress response pathways. Our species results contrasted with results from the previous study, indicating that further research is needed to improve the robustness of the results. Additionally, we found that offspring of hatchery and wild pallid sturgeon demonstrate different responses to heat stress, and potentially general stress that can occur in a hatchery environment. Overall, this study lays the groundwork for future research that can incorporate a larger suite of families to improve the robustness necessary to make actionable management recommendations.

Submitted in partial fulfillment for the requirements of the Degree of Doctor of Philosophy in the Department of Afrikaans in the Faculty of Arts at the University of Zululand, South Africa, 1999. / Engl: = The aim of this dissertation was to give an overview of work done on the grammatical expression of time in Afrikaans, and to show that the Cognitive Rhetoric, as a relatively new field of study, offers analytical tools that can account for problems being encountered with time expression which cannot be successfully dealt with through traditional methods of analysis and description. A descriptive framework was developed based on the cognitive approach of mental spaces, which has as basic principle the view that the semantic structure of a language is a reflection of a person's conscious experience of a physical, social and linguistic world, which allows for time to be metaphorically viewed as space. Thus time can be viewed as a container in which past, present and future are locked up in still smaller containers. These containers function as mental spaces, which are partial renderings of logical, coherent situations or potential realities in which time moves as a mover or a manipulator. An important tenet of the analytical framework which has been developed, is that a mental space for the present tense is regarded as man's primary field of experience, and as the cognitive point of departure from where he metaphorically projects to a past tense mental space or a future tense mental space. Within a particular mental space there is contained a sequence of events which are marked for the same tense; when a shift in time occurs, a new mental space is opened for the new time frame. The mental spaces approach has been utilized to show how time is used as a background-organizing mechanism for the analysis and description of tense in everyday discourse as well as a fictional narrative text. Afrik: = Die doelwit van hierdie dissertasie was om deur middel van bronverkenning 'n oorsig te bied van werk wat in Afrikaans gedoen is oor tydsvergestalting, en om aan te dui dat die Kognitiewe Retoriek, as opkomende studiegebied, ontledingsmeganismes bied vir probleme met tydsmarkering wat nie deur tradisionele metodes van ontleding en beskrywing opgelos kan word nie. 'n Ontledingsraamwerk gebaseer op die kognitiewe dinkruimbenadering is ontwikkel wat voorsiening maak vir die beginsel dat die semantiese struktuur van 'n taal 'n mens se bewussyn van 'n fisiese, sosiale en taalkundige wereld reflekteer, sodat tyd metafories as ruimte herbedink word. Hiervolgens word tyd as ln houer gesien waarbinne tydskonsepte soos hede, verlede en toekoms as kleiner houers opgesluit le*. Hierdie houers funksioneer as dinkruime, wat gedeeltelike weergawes is van logiese, samehangende situasies of potensiele werklikhede waarbinne tyd as 'n beweger en *n manipuleerder optree. *n Basiese vertrekpunt van die ontledingsraamwerk wat ontwikkel is, behels dat 'n dinkruim vir die teenwoordigetyd as die prim ere belewingsruim van die mens beskou word, en as die kognitiewe vertrekpunt vanwaar hy metafories projekteer na *n verledetyddinkruim of 'n toekomstigetyddinkruim. Binne 'n bepaalde dinkruim is daar 'n sekwensie van gebeurerame wat vir dieselfde tyd gemerk is; sodra *n tydskuif plaasvind, word 'n nuwe dinkruim oopgemaak vir die nuwe tydraam. Die dinkruimbenadering is benut om aan te toon hoe tyd as agtergrond-organiserende meganisme gebruik word om tyd in alledaagse diskoers asook in narratiewe tekste te ontleed. / University of Zululand and RGN.

Dans les cancers, la transcription des gènes peut être altérée par des mécanismes génétiques ou épigénétiques. En 2011, mon laboratoire a montré que la progression des cancers de la vessie pouvait être liée à un mécanisme épigénétique appelé MRES (« Mutiple Regional Epigenetic Silencing »). Les tumeurs possédant ce phénotype présentent une inactivation simultanée de gènes voisins dans 7 régions du génome. A l’aide d’une nouvelle approche bio-informatique : « SegCorr », nous avons identifié plus de 400 régions du génome dont l’expression des gènes est corrélée indépendamment du nombre de copie du gène. Ces régions se répartissent en 7 groupes et sont associées à 6 phénotypes de cancer de la vessie. De plus, l’extinction de l’expression des gènes d’une faible proportion de régions est associée à la méthylation de l’ADN et/ou une perte sur les histones de de marques d’activation de la transcription : H3K9ac et H3K4me3 ou des gains de marques de répression de la transcription : H3K27me3 et H3K9me3. Grâce au nouvel algorithme « Musette », J’ai montré que le phénotype MRES n’était probablement pas dû à des altérations génétiques. Enfin, pour comprendre à quel stade de la progression tumorale du cancer de la vessie le phénotype MRES pourrait apparaître, j’ai montré que les tumeurs de cancer de vessie induites chez les souris par ingestion d’un carcinogène (N-butyl-N-(4-hydroxybutyl)nitrosamine) pouvait être un bon modèle d’étude. / In cancers, gene transcription can be altered by genetic or epigenetic mechanisms. In 2011, my laboratory showed that the progression of bladder cancer could be linked to an epigenetic mechanism called MRES ("Mutiple Regional Epigenetic Silencing"). Tumors with this phenotype exhibit simultaneous inactivation of neighboring genes in 7 regions of the genome. Using a new bioinformatics approach: "SegCorr", we have identified more than 400 genomic regions in which gene expression is correlated. These regions fall into 7 groups and are associated with 6 phenotypes of bladder cancer. In addition, the extinction of gene expression from a small proportion of regions is associated with DNA methylation and / or loss of histone marks associated with active transcription: H3K9ac and H3K4me3 or gains of histone marks associated with transcription repression: H3K27me3 and H3K9me3. Using a new algorithm “Musette”, I have shown that the MRES phenotype is probably not due to genetic alterations. Finally, to understand at which stage of tumor progression of bladder cancer the MRES phenotype might appear, I have shown that bladder cancer tumors induced in mice by ingestion of a carcinogen (N-butyl-N-(4-hydroxybutyl) nitrosamine) could be a good model.

Although the subject of pottery would appear to the layman to be one of complicated formulas and difficult technical requirements, it is possible to reduce the on- tire porcedure to a series of simple, logical processes. To do this is the purpose for which this thesis has been undertaken. It seems advisable to reduce the entire subject of elementary pottery to fifteen lessons, for two reasons. (1) The instructor who must teach the subject in one se- mester of eighteen weeks will be enabled by this plan to cover the subject logically and completely if she will adhere to the divisions which have been made. This will give ample time for outside work, tests, etc. (2) The craftsman who wishes to express himself in one more medium, or the home-maker who merely desires the diversion of self-expression, will find the directions, given in this logical sequence, so clear and untechnical that he cannot fail, with sufficient practice, to achieve definite results.

Fertility is the most important factor controlling mammalian reproduction. Bull fertility, ability of the sperm to fertilize and activate the egg, and support embryonic development is crucial for early development. Similarly, the hormonal environment of the embryo also plays a critical role in successful embryonic development. We know that molecular health of the sperm as well as progesterone enhancing the development of the embryo is important for early development and implantation. The gaps in the knowledge base are 1) how early mammalian embryo development is paternally affected is not fully clear and 2) how progesterone improves the survival of the transferred embryo in the uterus still remains elusive. The central hypothesis was that low expression of MHC1 and Magel2 would cause a dysfunction in early embryo development and that progesterone will increase the survivability of the embryo via its associations with TNF-alpha and PGF2alpha. The objectives of this study were to 1) determine the expression of mhc1 and magel2 in single in vivo and in vitro blastocysts derived from low fertility and high fertility sires as well as to determine main pathways by which protein products of these genes regulate early development, and 2) identify the role of progesterone in regulating TNF-alpha and PGF2alpha. To accomplish these goals, we performed real time reverse transcriptase reaction and bioinformatics approaches, and ELISA and commercially available radioimmunoassay kits, respectively. The results of the experiments showed that 1) expression of mhcI transcripts were greater in high fertility in vivo embryos compared to their low fertility in vivo counterparts. Magel2 results showed an increase (P ≤ 0.05) in expression levels of high fertility in vivo embryos compared to their high fertility in vitro counterparts. Low fertility in vivo embryos had higher expression than high fertility in vitro embryos as well.

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