Testing SSA4::ADE3 Reporters for MCS Screening

Gene expression is essential to life and occurs through the processes of transcription of mRNA in the nucleus, export of transcripts to the cytoplasm through the nuclear pore complex (NPC), and translation of the mRNA into protein in the cytosol. The budding yeast S. cerevisiae is a eukaryotic model system used to explore the regulation of mRNA export. Transcripts are able to exit the nucleus through interaction with Mex67 which binds mRNA via adaptor proteins and allows crossing through NPCs. However, during heat shock (42°C) known adaptor proteins are rendered dysfunctional thus halting general mRNA export. Under these conditions, specific transcripts are able to exit the nucleus to enable the cell to response to stress. For example, SSA4 encodes a chaperone, which helps denatured proteins refold following heat shock is able to selectively export in response to stress. However, the mechanism for this selective export is not understood. I hypothesize that an unknown adaptor protein recruits Mex67 to allow for its selective export from the nucleus. In order to test this hypothesis, I have cloned a pair of vectors to be used as phenotypic reporters for SSA4 export. These reporters include the ADE3 ORF and SSA4 regulatory sequence (promoter, 5’ and 3’ UTRs). We anticipated that these reporters would yield red yeast cells if selectively exported during heat shock conditions. Analysis of the first series of reporter plasmids indicated that the plasmids weren’t fully cloned as expected, and subsequent experiments have been aimed at generating the proper clones. Future analysis will test whether these reporters successfully recapitulate selective SSA expression and export and use of these reporters for genetic screening to identify proteins involved in selective mRNA export

Unraveling the Role of Glycolysis-Derived Pyruvate in Modulation of TIM-3 Expression in Cancer

T cell exhaustion is characterized by reduced effector function, impaired proliferation, altered cytokine production, and compromised immune responses, particularly in chronic infections and cancer. Immune checkpoint molecules binding to exhaustion markers on T cells, including PD-1 and TIM-3, resulting in metabolic energy depletion, ultimately reducing immune-mediated clearance of tumor cells. Cellular protein expression is often resultant of chromatin accessibility to transcription factors (TFs), where accessible chromatin binding regions allow for TFs to bind and modulate protein expression, dictating cellular behaviors. Through prior flow cytometry analyses, the Chang Lab has found an association between elevated pyruvate or hypoxia levels and enhanced expression of the exhaustion marker TIM-3 in CD8+ T cells. We hypothesized that pyruvate accumulation in CD8+ T cells results in a signaling cascade that ultimately results in elevated TIM-3 expression and is correlated with cellular exhaustion. In this project, we found that pyruvate-induced TIM-3 expression is sensitive to metabolic inhibitors, further compounding the influence of metabolism in dictating T cell exhaustion phenotypes. Furthermore, we sought to better understand the signaling network that associates pyruvate accumulation, TIM-3 expression, and T cell exhaustion. Utilizing in vitro and molecular biology approaches, we have unraveled the complexity of this signaling pathway; identifying the importance of histone modification and JunB, JunD, and Bach2 TFs as critical regulators of pyruvate-induced TIM-3 expression. Collectively, these results provide significant insight into the role of pyruvate in modulating CD8+ T cell exhaustion

Fostering a Trauma Informed Mindset in the Criminal Justice System: An Evaluation of the Take Care Delaware Program

Fostering a Trauma Informed Mindset in the Criminal Justice System Lauren Ledford, Mary Jo Oliver M.S., Meredith Jordan, Chloe Salyer, Adam Walker, Andrea D. Clements Ph.D. The presence of early childhood adversity and trauma is disproportionately high in individuals involved with the criminal justice system compared to the non-criminally involved population. Childhood adversity and trauma is consistently linked to many adverse lifelong outcomes such as increased mortality, increased risk of mental illness, and increased risk of substance use disorder. In order to respond to this increased risk, empathy and community-wide interventions have emerged as effective means of responding and resisting re-traumatization. Community and empathy-based training of law enforcement officials may assist in understanding trauma and combatting its effects as they are often some of the first people to come in contact with individuals experiencing the effects of childhood adversity and trauma. This study sought to evaluate the effectiveness of Trauma-Informed Care (TIC) training for police officers in their ability to have knowledge of, believe to be relevant, understand, feel confident implementing, and apply TIC principles. We hypothesized knowledge, relevance, understanding, implementation confidence, and application of TIC would all increase from pre-assessment to post-assessment. As part of an initiative to increase TIC awareness and implementation, 64 police officers participated in a one-day training that began with a pre-survey and ended with a post-survey. Both surveys contained measures concerning TIC knowledge, relevance, understanding, implementation confidence, and application. The post survey also included an additional measure regarding training quality satisfaction. The sample consisted primarily of White (N = 44), middle-aged males (M = 42.27, SD = 8.588). There was a significant difference between TIC knowledge t(56) = -4.593, p\u3c.001, d = 1.096, TIC relevance t(56) = -2.488, p = .016, d = .912, TIC understanding t(56) = -8.611, p \u3c.001, d = 2.932, and implementation confidence t(56)= -5.942, p \u3c.001, d = 1.326 from the pre to post assessment. However, there was not a significant difference between TIC application t(56) = -1.826, p = .073, d = .585 from pre to post assessment. The consistently significant increase in TIC scores indicate that a TIC training is an effective means of improving officer perception of trauma and willingness to implement TIC practices