Determinants of Specificity for the Trypanosoma brucei A to I tRNA Editing Deaminase

Trypanosomatids are responsible for causing illness and death among millions of humans and animals worldwide. Two main genera of these unicellular flagellated parasites are Trypanosoma, which cause African sleeping sickness and Chagas disease, and Leishmania, which cause Leishmaniasis. The World Health Organization estimates that approximately 400 million people across the globe are at risk of infections caused by these protozoa. In addition to their pathogenic characteristics, trypanosomatids also represent some of the earliest mitochondria-containing eukaryotes. Trypanosomatids exhibit many significant and interesting biological processes, such as an extreme degree of RNA editing (as with U insertion and deletion in mRNAs) and complete tRNA import into the mitochondria, which is important for mitochondrial protein synthesis. In translation, although there are 61 amino acid codons, there is only a small subset of tRNAs to decode them. As a result, some flexibility is needed to allow for a single tRNA to decode multiple codons—as proposed by Francis Crick with the Wobble hypothesis in the 1960s (1). One nucleotide capable of increasing pairing flexibility during decoding is the nucleotide inosine, which in the third position of the tRNA anticodon, allows recognition of up to three different nucleotides in an mRNA codon: uridine, cytosine, and adenosine. The editing event that leads to inosine formation in tRNAs, is catalyzed by a heterodimeric enzyme known as ADAT2/3 (adenosine deaminases acting on tRNA). Remarkably, inosine is not encoded in DNA and is only found in RNAs as a result of an enzyme-mediated reaction, which is essential for viability. The goal of this project was to analyze the effects various mutations at the anticodon stem loop of tRNAThr have on inosine formation at the first (wobble) position of the anticodon of tRNAs in Trypanosoma brucei. We hypothesized that mutations at these positions of the tRNA will decrease deamination efficiency and substrate recognition by ADAT2/3. Through in vitro methods, which include protein-RNA binding assays (band-shift assays) and enzyme kinetics the effect of mutations on ADAT2/3 function was evaluated. These results could shed light on how ADAT2/3 binds tRNA and catalyzes the conversion of adenosine to inosine. In addition, these observations can also lead to future design of therapeutics against parasites of such major medical importance (2). 1. Crick, FH. (1966). Codon—anticodon pairing: the wobble hypothesis. J Mol Biol. 19, 548-555. 2. Gerber, A. P., & Keller, W. (1999). An adenosine deaminase that generates inosine at the wobble position of tRNAs. Science. 286, 1146-49.Dean's Undergraduate Research FundCollege of Arts and Sciences Research ScholarshipAmerican Heart AssociationNational Science FoundationNo embarg

Implementation of a Multi-Pronged Approach to Improve Education in Quality Improvement (QI) and to Increase Resident Involvement in QI Work

Objective: To evaluate a multi-tiered approach to incorporating QI and PS training and participation in a pediatrics residency program. Methods: We developed a multi-tiered approach to QI/PS training during the 2013-14 year, and expanded upon it during subsequent years. We tracked resident participation in Departmental QI Grand Rounds and ACGME and Department survey results. The initial intervention involved didactic and small group sessions/to facilitate project development. We also instituted Departmental QI Grand Rounds. In the following year (2014-15), we increased the number of didactics and started a practice improvement project within the resident group practice where residents were provided individualized practice data. In 2016-17, we added an adapted Team-Based Learning session for PGY-1’s and all residents were required to complete on-line Institute for Healthcare Improvement modules. Results: In 2013/14, 26 residents presented 2 oral and 12 poster presentations at QI Grand Rounds. This increased to 36, 2 & 14 in 2014/15, 43, 3 & 17 in 2015/16, and 26, 2 & 13 in 2016/17. In 2012/13, 50% of residents responded affirmatively to the ACGME survey question \u27participated in QI\u27; (19% affirmed they were \u27provided data about practice habits\u27). This increased to 86(29) in 2013/14; 81(48) in 2014/15, 83(42) in 2015/16, and 78(48) in 2016/17. Conclusions: We have implemented a multi-pronged QI curriculum that includes all resident levels. Results demonstrate increased resident involvement. Key to buy-in is the ability to participate in high-interest projects within self-designated tracts. Future directions include further study of the curriculum and of the system/patient safety outcomes

Setting Foundations for Pediatric Fellowship Core Curriculum Training

Purpose and background: The American Board of Pediatrics (ABP) and the Accreditation Council for Graduate Medical Education (ACGME) require pediatric subspecialty fellows to learn a core curriculum of scholarly topics including biostatistics, research methodology, teaching, quality improvement and leadership. Traditionally, each fellowship would develop such curricula for their own fellows –leading to variable experiences for fellows in different disciplines. Nationally, many fellowships are working to provide a common core curriculum which is available across disciplines with an aim towards improving training and transitioning to academic careers.1,2,3 To address this, we have developed a core curriculum which aims to provide all pediatric subspecialty fellows appropriate, expert, and timely education on these and other topics for their professional development. Learner Objectives: By completion of the Pediatric fellows core curriculum, pediatric fellows will be able to: Recognize the requirements set out by the ABP and ACGME for fellowship training. Recognize principles of scholarly activity in research and quality improvement. Describe principles and skills for teaching learners in different settings

Quiet Please: The effects of sleep quality and quantity as a result of a QI project to minimize nighttime sleep interruptions

Quiet Please! The effects on sleep quality and quantity as a result of QI project to minimize nighttime interruptions INTRODUCTION: Sleep is a significant component of health that is often overlooked in hospitalized patients. We introduced a QI project in 2016 to minimize nighttime interruptions in clinically stable patients by performing “passive vitals” which eliminated temperature and blood pressure readings at 4am and allowed patients to sleep longer overnight. Despite this intervention, the effects on sleep quality and quantity are unknown. OBJECTIVE: The objective was to compare total sleep time, sleep interruptions, and sleep quality between patients eligible for passive vitals and those ineligible through survey data collected from July to November 2019. METHODS: Participants eligible and non-eligible for passive vitals self-reported the number of hours of sleep, number of nighttime awakenings, awakenings attributed to hospital staff, and elaborated on reasons for poor sleep. The survey also allowed previously hospitalized patients to compare the current stay to those in the past by assessing their quality of sleep and how tired they felt upon waking using a 5-point scale. Data was analyzed using Chi-square test. RESULTS: Forty-eight eligible and fifty-six non-eligible patients were surveyed. The total hours of sleep between groups was not statistically significant (p=0.11). There were no significant differences in the number of total sleep interruptions (p=0.95) and those by hospital staff (p =0.55). For patients who were previously hospitalized, there were no significant differences in sleep quality rating (p=0.10) and how tired they felt the next morning (p=0.78). Both groups cited similar reasons for poor sleep including care-related and illness symptom-related disruptions most commonly, followed by environmental-related complaints. DISCUSSION: The total hours of sleep, number of sleep interruptions, sleep quality, and rating of tiredness did not differ among the eligible and non-eligible groups. Many factors contributed to poor overall sleep which cannot be mitigated by passive vital signs in eligible patients at 4am. Other factors will need to be addressed

RAM$mart Financial Wellness for Health Profession Students

College students today are graduating with more debt than ever before. As such, it is critical that our graduates are equipped with the right skills and frame of mind to manage their finances. This includes understanding their income, taxes, and expenses; managing debt; budgeting; saving; and retirement planning. Money management can be intimidating for anyone, regardless of background or education. While online resources are made available to our students, these resources are often overlooked, and do not necessarily set out to engage students or address the issues they may face in a convincing or compelling way. Most of the financial resources easily accessible on the web and on VCU’s sites are intended for undergraduate students. However, graduate students and in particular students in the health sciences professional programs (medicine, dentistry, pharmacy) can accrue a tremendous amount of debt, but are not exposed to as many mandatory or voluntary budgeting and financial resources. The RAM$mart Financial Wellness Program is a set of online modules that will allow students the opportunity to learn the basic principles of money and debt management. We are focusing this phase on professional students in the Schools of Medicine, Pharmacy, and Dentistry, with the intent to engage the whole VCU student population in the future. The RAM$mart Financial Wellness Program will include modules on a variety of money management topics designed to get the attention of students and to provide them with practical steps they can take at the beginning of, and throughout, their graduate professional education to minimize debt accumulation. There will also be modules designed to inform students of various loan payback options upon their degree completion, and of available community and online financial resources. Through attention-grabbing and entertaining modules, including such things as humorous video snippets and interactive questions, students’ awareness of budgeting and financial responsibility will increase significantly

Improving Early Antibiotic Administration for Treatment of Sepsis at Children’s Hospital of Richmond at VCU: 2012-2019

Background: The Surviving Sepsis Campaign recommends initiating IV antibiotic administration within one hour of recognition of severe sepsis. Several studies have shown that prompt blood culture collection, administration of broad-spectrum antibiotics, and fluid resuscitation following recognition improves child survival. Objective: Our goal was to evaluate effectiveness of sepsis initiatives and institutional changes in the timing of early antibiotic administration at Children’s Hospital of Richmond at VCU. Methods: We formed a Pediatric Sepsis Committee with representatives from each unit in 2013. In 2016, the committee began tracking time from the order of a first stat dose IV antibiotic to administration as a marker of early treatment and reviewed data on a monthly basis with run charts for overall and unit-specific data. Other interventions included improved availability of antibiotics in automated dispensing machines, sepsis screening and alert systems, sepsis huddles, and auto-generated pages to charge nurses upon order of stat IV antibiotics. We included percent of stat antibiotics administered in less than one hour from order across all pediatric units since 2012. Results: Across all units, the centerline of first dose stat antibiotics delivered within one hour improved from a baseline of 34% in 2012 to 76% in 2019. The NICU and PICU centerlines improved by 53% and 48%, respectively since 2012. The Pediatric ED improved from 66% in 2012 to 84% in 2016. The Acute Care Pediatrics (ACP) Unit centerline improved from 24% in 2012 to 50% in 2017. Conclusion: Time from order to stat antibiotic administration has improved in all units receiving quality improvement initiatives. These improvements have been made possible by widespread emphasis on the dangers of untreated sepsis, multidisciplinary collaboration between nursing and physician staff, structural pharmacy changes and electronic alerts. Further studies are needed to determine impact on patient outcomes

Evidence that dopamine acts via Kisspeptin to Hold GnRH pulse frequency in check in Anestrous Ewes

Recent work has implicated stimulatory kisspeptin neurons in the arcuate nucleus (ARC) as important for seasonal changes in reproductive function in sheep, but earlier studies support a role for inhibitory A15 dopaminergic (DA) neurons in the suppression of GnRH (and LH) pulse frequency in the nonbreeding (anestrous) season. Because A15 neurons project to the ARC, we performed three experiments to test the hypothesis that A15 neurons act via ARC kisspeptin neurons to inhibit LH in anestrus: 1) we used dual immunocytochemistry to determine whether these ARC neurons contain D2 dopamine receptor (D2-R), the receptor responsible for inhibition of LH in anestrus; 2) wetested the ability of local administration of sulpiride, a D2-R antagonist, into theARCto increase LH secretion in anestrus; and 3) we determined whether an antagonist to the kisspeptin receptor could block the increase in LH secretion induced by sulpiride in anestrus. In experiment 1, 40% of this ARC neuronal subpopulation contained D2-R in breeding season ewes, but this increased to approximately 80% in anestrus. In experiment 2, local microinjection of the two highest doses (10 and 50 nmol) of sulpiride into the ARC significantly increased LH pulse frequency to levels 3 times that seen with vehicle injections. Finally, intracerebroventricular infusion of a kisspeptin receptor antagonist completely blocked the increase in LH pulse frequency induced by systemic administration of sulpiride to anestrous ewes. These results support the hypothesis that DA acts to inhibit GnRH (and LH) secretion in anestrus by suppressing the activity of ARC kisspeptin neurons.We thank Heather Bungard and Jennifer Lydon (West Virginia University Food Animal Research Facility) for the care of animals and Paul Harton for his technical assistance in sectioning tissue. We also thank Dr. Al Parlow and the National Hormone and Peptide Program (Torrance, CA) for the reagents used to measure LH and prolactin.http://endo.endojournals.org/am201