Development of novel hybridization assays and immunoassays based on bioluminescence.

The ability of labeled DNA or RNA probes to bind with high affinity and specificity to complementary nucleic acid sequences forms the basis of nucleic acid hybridization assays. Immunoassays, on the other hand, exploit the strong and specific interaction between antigens and antibodies. One aspect of this dissertation deals with enhancement of the sensitivity of bioluminescence hybridization assays based on the photoprotein aequorin. This is achieved by introducing, enzymatically, multiple aequorin labels per DNA hybrid. The bound aequorin is determined by its characteristic bioluminescence. An 8-fold improvement in sensitivity is observed with the amplified assay as compared to an assay without the amplification step. Another aspect of this dissertation involves the development of two novel microtiter well-based DNA hybridization assays in which an expressible cDNA fragment encoding firefly luciferase serves as a reporter molecule. The reporter molecule undergoes in vitro expression through coupled transcription/translation or through sequential transcription/translation reactions. The bioluminescence activity of generated firefly luciferase was measured. The expression yield was improved with the separate transcription/translation protocol. In vitro expression provides signal amplification in hybridization assays because each cDNA label generates several enzyme molecules in solution. A third aspect of this dissertation combines the firefly luciferase cDNA and a cDNA encoding Renilla luciferase as labels for the development of a microtiter well-based expression hybridization assay that allows simultaneous determination of two target DNA sequences. The cDNA labels were expressed in vitro simultaneously and independently in the same transcription/translation reaction mixture. The activities of synthesized firefly and Renilla luciferases were co-determined in a single sample based on the requirements of their characteristic bioluminescent reactions. In another aspect of this dissertation, the concept of in vitro expression as an analytical tool was further expanded in a novel microtiter well immunoassay which uses T7 RNA polymerase itself as the label. The enzyme label was determined by using the bound T7 RNA polymerase for in vitro expression of a firefly luciferase-coding DNA fragment. The activity of synthesized luciferase was measured. This is the first time that T7 RNA polymerase serves as the reporter molecule in hybridization assays or immunoassays.Dept. of Chemistry and Biochemistry. Paper copy at Leddy Library: Theses & Major Papers - Basement, West Bldg. / Call Number: Thesis2000 .L35. Source: Dissertation Abstracts International, Volume: 61-09, Section: B, page: 4700. Adviser: Theodore K. Christopoulos. Thesis (Ph.D.)--University of Windsor (Canada), 2000

Using Infographics to Go Public with SoTL

There has been a call to amplify the scholarship of teaching and learning (SoTL) and expand its reach by engaging with audiences outside the academy. In this paper, we share our journey in crossing disciplinary boundaries and creating a SoTL-informed infographic for public consumption. As the field of SoTL continues to evolve, infographics hold tremendous potential to communicate SoTL to various stakeholders, including educators, students, administrators, policymakers, and the public. We outline best practices in infographic development and the potential of infographics as a tool for taking SoTL public, emphasizing their visual appeal and effectiveness in conveying complex information. We conclude by discussing the implications of using infographics to advance SoTL communication. The efforts of our group serve as a valuable example of how infographics can be used to bring SoTL knowledge out of academia and into the public domain

Public participation in healthcare students' education: An umbrella review

Abstract Background An often‐hidden element in healthcare students' education is the pedagogy of public involvement, yet public participation can result in deep learning for students with positive impacts on the public who participate. Objective This article aimed to synthesize published literature reviews that described the impact of public participation in healthcare students' education. Search Strategy We searched MEDLINE, EMBASE, ERIC, PsychINFO, CINAHL, PubMed, JBI Database of Systematic Reviews and Implementation Reports, the Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effects and the PROSPERO register for literature reviews on public participation in healthcare students' education. Inclusion Criteria Reviews published in the last 10 years were included if they described patient or public participation in healthcare students' education and reported the impacts on students, the public, curricula or healthcare systems. Data Extraction and Synthesis Data were extracted using a predesigned data extraction form and narratively synthesized. Main Results Twenty reviews met our inclusion criteria reporting on outcomes related to students, the public, curriculum and future professional practice. Discussion and Conclusion Our findings raise awareness of the benefits and challenges of public participation in healthcare students' education and may inform future research exploring how public participation can best be utilized in higher education. Patient or Public Contribution This review was inspired by conversations with public healthcare consumers who saw value in public participation in healthcare students' education. Studies included involved public participants, providing a deeper understanding of the impacts of public participation in healthcare students' education

Circadian cortisol profiles, anxiety and depressive symptomatology, and body mass index in a clinical population of obese children

Obesity is highly co-morbid with anxiety and/or depression in children, conditions that may further worsen the metabolic and cardiovascular risks for obese individuals. Dysregulation of the hypothalamic-pituitary-adrenal axis is involved in the pathophysiology of anxiety disorders, depression, and obesity, and diverse cortisol concentrations may be found in obese children, depending on their degree of psychological distress. The aim of this study was to examine cortisol profiles among obese children with or without symptoms of anxiety and depression. A group of 128 children (53% females; mean age +/- SD: 11.2 +/- 2.2 years) derived from a pediatric obesity clinic were studied. Anxiety and depressive symptomatology were assessed with appropriate instruments. Morning serum and five diurnal salivary cortisol concentrations were measured. Obese children were 3.1/2.3 times more likely to report state and trait anxiety, respectively, and 3.6 times more likely to report depressive symptoms than children of the same age group, from a contemporary Greek sample. Trait anxiety and noon salivary cortisol concentrations were significantly positively correlated (p = 0.002). Overall, salivary cortisol concentrations were increased in children with anxiety or depression symptomatology compared to obese children without any affective morbidity (p = 0.02) and to those with anxiety and depression co-morbidity (p = 0.02). In conclusion, in obese children, emotional distress expressed by symptoms of anxiety and/or depression is associated with circadian cortisol profiles reflecting a potential pathway for further morbidity. Longitudinal studies may reveal a role of cortisol in linking obesity, anxiety, and depression to the development of further psychological and physical morbidity