Method Development for the Assignment of NMR Peaks and the Determination of Quaternary Protein Structure using Reductive Methylation

The work described herein is centered on reductive methylation, a chemical modification reaction, which allows for the introduction of methyl groups at the primary amines of the lysine side chain and at the N-terminus. The resulting methyl groups are effective probes for the study of protein structure and dynamics using nuclear magnetic resonance spectroscopy (NMR). Described herein is work conducted on the development of tools for the study of protein structure using reductive methylation, highlighting options for addressing limitations of the labeling method encountered by its practical use. Chapter 1 serves as a brief introduction to protein structure and the modification of protein structure, both biological and chemical. This chapter sets the stage for the remaining chapters by establishing the importance of structural studies and how the reductive methylation reaction has been useful to this end. Chapter 2 details work done towards the development of a method to make assignments of NMR peaks of a reductively methylated protein. The method described focuses on addressing the issue of degenerate labeling associated with the reaction, which results in similar degrees of methylation at the reaction sites. The method presented involves the introduction of solid chromatographic media into the reaction and using substoichiometric concentrations of the modifying reagents in order to introduce more variation in the labeling of the reaction sites. The overall objective of this project is to investigate if the introduction of the media and the change in reagent concentration would help to overcome the degenerate labeling and enable more definitive peak assignments. In chapter 3, methods were developed by modifying the sample environments to achieve more resolved and intense NMR peaks of reductively methylated protein. The improvement of resolution and intensity aids peak fitting for accurate peak area calculations, which are used in the determination of percent labeling at each dimethylamine peak in the NMR spectra. Chapter 4 details a study where the overall objective is to investigate the effects of reductive methylation on the determination of protein concentration by the Bradford and Bicinchoninic acid (BCA) assays. The development of a novel use of reductive methylation coupled with paramagnetic relaxation enhancement (PRE) NMR spectroscopy is detailed in Chapter 5 for the determination of the quaternary structural complex of a cysteine protease, papain, and a cysteine protease inhibitor. Chapter 6 will discuss the overall conclusions of this dissertation research and future directions of the work presented

A Colourful Approach To Reducing Examination Anxiety

This paper examines the use of visual aids in examinations and provides interesting and innovative ideas for educators in higher education institutions. It shows a clear link between the use of colour and technology and the reduction of examination anxiety. Traditionally examinations tended to be in written format only, questions were multiple choice, short answer, essay or case study based. This paper showcases an innovative approach to examinations where videos and photos are use to supplement traditional methods. A sound theoretical foundation is provided to support the use of the innovative methods utilized. The paper not only provides information for educators on how to approach this idea but more importantly documents student feedback on the successful implementation of the concept

Enhancing Learning Outcomes Through Application Driven Activities In Marketing

This paper introduces an activity used in class to allow students to apply previously acquired information to a hands-on task. As the authors have previously shown active learning is a way to effectively facilitate and improve students’ learning outcomes. As a result to improve learning outcomes we have overtime developed a series of learning activities reinforcing marketing theories. In this paper, we provide details of a marketing communications exercise which in addition to being a vehicle for active learning also allowed students to apply the results of their previous research thereby effectively aligning all learning and assessment tasks. The activity required students to take a previously completed environmental analysis of a foreign country and using that information develop a print advertisement for an Australian branded product unavailable in that country. Details are provided on the in-class activity including guidelines for conducting the practice-orientated exercise and information on the assessment tasks leading up to this activity. The paper provides academics with an example of how to apply concepts to real life examples in the classroom and give students hands on experience and further shows how learning can be scaffolded through assessment. We found that students actively engaged in the experience and improved their learning outcomes through application of their previous research and the concepts discussed in class

Putting Bling In The Classroom

This paper showcases the innovative use of a case study in action.  It initially introduces a case study on the bottled water ‘bling’ and its phenomenal success in recent times.  From a marketing educator’s point of view it provides an example of how to bring reality to the class room and gives students hands on consumer experience through active learning.  Details are provided on the in-class activity consisting of the case study, questions and guidelines for conducting the practice-orientated exercise.  The paper concludes with student feedback on the activity, which highlights the level of student engagement and motivation to learning and class participation.  It clearly shows that immersing students in the experience through active learning improves learning outcomes through concretisation of the concepts discussed in class. It also provides educators with recommendations for replicating this exercise

Innovating Through Structured Curriculum Development

Structured and innovative curriculum development can have a profound impact on learning outcomes.  This paper provides interesting and innovative ideas for educators in higher education institutions to allow them to achieve improved learning outcomes.  This paper showcases how engaging students throughout the course and supporting them through consistent, incremental course development leads to innovative curriculum development.  The improved course structure and students’ increased engagement achieve maximum learning outcomes.  Various research studies support the use of innovative methods though not much is said about consistency of assessments and course structure in curriculum development with the aim to reduce exam anxiety and ultimately enhance learning outcomes.  In our unit of study, seminar presentations and poster sessions in conference style were used to engage students and introduce them to a different way of learning other than reports.  The paper not only provides information for educators on how to design engaging course structures but more significantly supports the outcomes through the analysis of student feedback based on objective teaching evaluation conducted before exams

Insights and evidence gaps in girl-centered programming: A systematic review

Increased attention to the needs of adolescent girls has led to a growing number of programs in low- and middle-income countries. Questions remain, however, about what aspects of program design are most effective. This hinders efforts to effectively allocate resources, scale up programs, and replicate results across settings. This review looks at how the number of program components, involvement of supporting actors who influence the lives of girls, supplemental “booster” activities, intervention exposure level, and community saturation level influenced outcomes for girls. While findings suggest the importance of multicomponent programs and longer program exposure, each area requires further rigorous research to determine whether and under what circumstances they amplify impact

Three-Dimensional Paper-Based Microfluidic Device for Assays of Protein and Glucose in Urine

The first step in curing a disease is being able to detect the disease effectively. Paper-based microfluidic devices are biodegradable and can make diagnosing diseases cost-effective and easy in almost all environments. We created a three-dimesnional (3D) paper device using wax printing fabrication technique and basic principles of origami. This design allows for a versatile fabrication technique over previously reported patterning of SU-8 photoresist on chromatography paper by employing a readily available wax printer. The design also utilizes multiple colorimetric assays that can accommodate one or more analytes including urine, blood, and saliva. In this case to demonstrate the functionality of the 3D paper-based microfluidic system, a urinalysis of protein and glucose assays is conducted. The amounts of glucose and protein introduced to the device are found to be proportional to the color change of each assay. This color change was quantified by use of Adobe Photoshop. Urine samples from participants with no pre-existing health conditions and one person with diabetes were collected and compared against synthetic urine samples with predetermined glucose and protein levels. Utilizing this method, we were able to confirm that both protein and glucose levels were in fact within healthy ranges for healthy participants. For the participant with diabetes, glucose was found to be above the healthy range while the protein level was in the healthy range

Higher quality quit-date goal setting enhances quit attempts among quitline callers

INTRODUCTION At tobacco quitlines, coaching and cessation medications are commonly structured around setting a date for making a quit attempt. However, limited literature evaluating this practice suggests that callers do not routinely set quit-date goals. High quality goal setting may increase the frequency of caller quit attempts. In this study, we examine the quality of quit-date goal setting and its association with in-program quit attempts and the timing of callers' first quit attempt. METHODS Using call recordings, we scored the quality of quit-date goal setting among 90 callers enrolled at Arizona Smokers' Helpline between August and December 2017. The primary exposure was quality of quit-date goal setting assessed using the Lorencatto et al. rating scale. Coding reliability was assessed using Cohen's kappa. Multivariable logistic regression was used to examine the association between quality of goal setting and in-program quit attempts (>24 h tobacco free). RESULTS The mean quality goal setting score was 3.1 (range: -3 to 7). Sixty-nine callers (77%) set a quit date and 39 (43%) made a quit attempt. Compared to callers who experienced low-quality goal setting, the adjusted odds of in-program quitting for high quality goal setting was AOR=3.98 (95% CI: 1.55-10.20) and for making a quit attempt within two weeks OR=6.23 (95% CI: 1.52-25.49). CONCLUSIONS Quit-date goal setting is an important element of quitline services and callers benefit from high quality quit-date goal setting. Quitlines should establish quality improvement measures to ensure that coaches are trained to provide high quality quit-date goal setting opportunities to all callers.Arizona Department of Health Services [ADHS11-007339, ADHS16-106672, ADHS13-026130:5]; NIH-NCI Cancer Center [P30 CA023074]Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Electronic cigarette use and tobacco cessation in a state-based quitline

Introduction. Evidence is mixed on e-cigarette's effectiveness as a tobacco cessation aid. Research suggests that e-cigarette users face greater barriers to quitting tobacco. Aim. To examine the association between e-cigarette use and tobacco cessation outcomes among quitline callers. Methods. We examined 2,204 callers who enrolled and completed 7-month follow-up surveys between April 2014 and January 2017. We examined the association between any e-cigarette use and tobacco cessation. We also evaluated these relationships by e-cigarette use patterns between enrollment and 7-month follow-up: sustained, adopted, discontinued, and non-use. We used multivariable logistic regression to control for caller characteristics, tobacco history, and program utilization. Results. Overall, 18% of callers reported using e-cigarettes at enrollment, follow-up, or both. Compared to non-users, e-cigarette users were more likely to be younger, non-Hispanic, and report a mental health condition. The adjusted odds of tobacco cessation were not statistically different for callers who used e-cigarettes compared to those who did not (adjusted odds ratios = 1.02, 95% confidence interval 0.79-1.32). Results were similar when examining cessation by patterns of e-cigarette use. Conclusions. E-cigarette use was not associated with tobacco cessation. This suggests that e-cigarette use may neither facilitate nor deter tobacco cessation among quitline callers. Future research should continue exploring how e-cigarette use affects quitting.6 month embargo; published online: 20 March 2019This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Human Fear Conditioning Conducted in Full Immersion 3-Dimensional Virtual Reality

Fear conditioning is a widely used paradigm in non-human animal research to investigate the neural mechanisms underlying fear and anxiety. A major challenge in conducting conditioning studies in humans is the ability to strongly manipulate or simulate the environmental contexts that are associated with conditioned emotional behaviors. In this regard, virtual reality (VR) technology is a promising tool. Yet, adapting this technology to meet experimental constraints requires special accommodations. Here we address the methodological issues involved when conducting fear conditioning in a fully immersive 6-sided VR environment and present fear conditioning data