Generation of a CCL2 knockout using CRISPR/CAS9 and lipid mediated transfection in CT-26 murine colon carcinoma cells

CCL2 is an inflammatory mediator that is released by tumor cells to activate and direct immune cell species, especially macrophages, to inflammatory sites within the body. The goal of this project was to successfully generate knockout the CCL2 ligand gene using a CRISPR/Cas9 complex delivered via lipid mediated transfection. The sgRNA and Cas9 mRNA were introduced into the cells via lipid-mediated transfection. The cells were incubated for 4 days, before being analyzed using PCR and gel electrophoresis. We expected to see one band on the first gel and two bands on the second gel. Two bands appeared on the first gel and 2 bands appeared on the second gel. This indicates that the target sequence was amplified, but the transfection efficiency was too low for the enzyme to detect cleavage. We conclude that lipid mediated transfection is an efficient method for introducing foreign genetic material into CT-26 cells

Transcriptional repression by ApiAP2 factors is central to chronic toxoplasmosis

Tachyzoite to bradyzoite development in Toxoplasma is marked by major changes in gene expression resulting in a parasite that expresses a new repertoire of surface antigens hidden inside a modified parasitophorous vacuole called the tissue cyst. The factors that control this important life cycle transition are not well understood. Here we describe an important transcriptional repressor mechanism controlling bradyzoite differentiation that operates in the tachyzoite stage. The ApiAP2 factor, AP2IV-4, is a nuclear factor dynamically expressed in late S phase through mitosis/cytokinesis of the tachyzoite cell cycle. Remarkably, deletion of the AP2IV-4 locus resulted in the expression of a subset of bradyzoite-specific proteins in replicating tachyzoites that included tissue cyst wall components BPK1, MCP4, CST1 and the surface antigen SRS9. In the murine animal model, the mis-timing of bradyzoite antigens in tachyzoites lacking AP2IV-4 caused a potent inflammatory monocyte immune response that effectively eliminated this parasite and prevented tissue cyst formation in mouse brain tissue. Altogether, these results indicate that suppression of bradyzoite antigens by AP2IV-4 during acute infection is required for Toxoplasma to successfully establish a chronic infection in the immune-competent host

Investigating the cognitive mechanisms by which sleep supports emotion regulation and mental health

Sleep plays an important role in how we process and deal with our emotions on a daily basis. As emotion regulation difficulties are a key predictor of poorer mental health, understanding the mechanisms by which sleep supports emotion regulation and mental health is of the upmost importance to further our understanding of psychiatric vulnerability. This thesis aims to investigate the cognitive mechanisms by which sleep supports emotion regulation and mental health. Specifically, three components of emotion regulation are examined: cognitive emotion regulation (CER), emotional reactivity, and emotional inertia. The first empirical chapter (Chapter 2) investigates whether the benefits of adaptive CER strategies (to lower depression and anxiety) are contingent on high sleep quality. The second empirical chapter (Chapter 3) examines whether sleep deprivation (versus a night of sleep) influences the evolution of arousal responses during exposure to ambiguous threat, as well as the reciprocal influence of slow wave activity (SWA) on affect regulation. The third empirical chapter (Chapter 4) explores whether the benefits of adaptive CER strategy use (to lower emotional inertia) are contingent on high sleep quality. Our findings suggest that: 1) greater use of adaptive CER strategies and high sleep quality independently promote resilience to depression, 2) a night of sleep (versus sleep deprivation) promotes the regulation of affect in response to prolonged ambiguous threat; however, SWA is not associated with this regulation, and 3) greater use of adaptive CER strategies and high sleep quality independently reduce the persistence of negative emotions over time. In light of these findings, cognitive control is proposed as one critical mechanism underlying the association between sleep and emotion regulation. Altogether, this thesis provides important insights into the cognitive mechanisms by which sleep supports emotion regulation, and mental health, and points towards modifiable mechanisms that may buffer against psychiatric vulnerability

Executive function and novel word learning in monolingual and bilingual children

The proposed study aimed to compare the executive function ability of monolingual and bilingual preschoolers between 4 and 6 years. The preliminary data suggests that both groups differed across all the tasks except for the working memory subcomponent. Although both working memory tasks were visuospatial and assessed the same subcomponents, there were discrepancies in results due to the task setup. Researchers and clinicians must be cautious while selecting and setting the tasks as tasks employed could bring about performance differences

Positron scattering from molecular hydrogen

We present results for total and partial cross sections for positron scattering from H2. The total scattering and positronium formation cross sections are reported between 0.5 and 200 eV. Total quasielastic and inelastic scattering cross sections are rep

Comic license and extreme figurations in contemporary American storytelling

In this project I examine the significance and sophistication of comedy in contemporary American storytelling, in order to get beyond its frequent characterisation as either a superficial sweetener or a form of escapism. My thesis argues instead for comedy’s usefulness as a strategy for effecting multiple responses: intimacy, recognition, attachment, de-familiarisation, celebration and catharsis, all in the service of confronting the unbearable. Examining work by George Saunders, Miranda July, Donald Antrim, and the filmmaker, Jordan Peele, I argue that comic license allows for the forthright address of troubling issues; class in Saunders’s short fiction; sexuality in July’s first novel; national identity for Antrim and race in Peele’s film, Get Out. This license is effected in multiple ways: through the obliviousness of characterological traits such as naivety and pedantry for instance, as well as through the mechanisms of incongruity and relief. Given that the works I examine are challenging, ‘edgy’, both in terms of style and content, I suggest that the comedy is inflected by what we might loosely call extremity. While extremity is arguably foundational to comedy, in that the comic violation of conventional boundaries necessitates it to one degree or another; I also argue that the comic license enables the work’s ‘extreme figurations’ by ensuring the reader’s or the viewer’s consent. Comic pleasure thus keeps us close to what might otherwise be overwhelming. In addition, comedy’s tendency towards the material and the particular – the proverbial slip on the banana skin - helps to ground or embed the extremity of the work’s abstract or fantastical aspect

The Effect of Viewing Appearance-Based and Self-Compassion Social Media Content on Body Image

The present study investigated the impact of viewing appearance-focused body-positive content versus non-appearance focused self-compassion content on a person’s self-image. Previous studies have demonstrated that viewing body-positive content improves body self-esteem. Additionally, self-compassion content may be a protective factor against body dissatisfaction. Although body-positive content is associated with increased body self-esteem, some question remains whether the continued focus on appearance is an effective strategy. Therefore, the purpose of the current study is to examine whether non-appearance based content provides benefits above and beyond those provided by body positivity. It was hypothesized that those who viewed non-appearance based content would report greater positive effects on body self-esteem than those who view appearance focused content, as non-appearance focused content directs the viewer\u27s attention away from self-objectifying tendencies. Covariation effects of ideal internalization and trait appearance comparison characteristics were also investigated. Both experimental conditions demonstrated a significant increase in measures of body self-esteem, however they did not differ significantly from each other. Ideal internalization was found to be a significant covariate. Exploratory gender analyses indicated that while both types of positive content significantly increased body self-esteem in both men and women, appearance-neutral content may have a stronger effect for men. Results of this study provide further evidence for the effectiveness of including both appearance-based and appearance-neutral content in social media interventions for increasing positive body image. Additionally results suggest that gender differences should be continued to be investigated in future research as varying the type of content may enhance effectivity by gender

Using Robots to Promote Social and Emotional Learning

Claire Sullivan ’21Major: Elementary and Special Education Maya Young ’21Major: Elementary and Special Education Emma Hendricks ’21Major: Elementary and Special Education Faculty Mentor: Dr. Lin Zhang, Elementary and Special Education Technology based interventions, such as computer programs and virtual reality, generally yield positive results when used with students with autism spectrum disorders (ASD). It is speculated that these devices may be successful because of their novelty and because they do not require the student with ASD to be socially interactive with a person. This project builds on the existing body of research and uses Root robots to enhance the communication/collaboration skills of children with special needs, particularly those with autism, through social interactions