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Streptococcus pyogenes (S. pyogenes) can thrive in its host during an infection, and, as a result, it must be able to respond to external stimuli and available carbon sources. The preclinical use of engineered pathogens capable of constitutive light production may provide real-time information on microbial-specific metabolic processes. In this study, we mapped the central metabolism of a luxABCDE-modified S. pyogenes Xen20 (Strep. Xen20) to its de novo synthesis of luciferase substrates as assessed by the rate of light production in response to different environmental triggers. Previous characterization predicted that the lux operon was under the myo-inositol iolE promotor. In this study, we revealed that supplementation with myo-inositol generated increased Strep. Xen20 luminescence. Surprisingly, when supplemented with infection-relevant carbon sources, such as glucose or glycine, light production was diminished. This was presumably due to the scavenging of pyruvate by L-lactate dehydrogenase (LDH). Inhibition of LDH by its inhibitor, oxamate, partially restored luminescent signal in the presence of glucose, presumably by allowing the resulting pyruvate to proceed to acetyl-coenzyme A (CoA). This phenomenon appeared specific to the lactic acid bacterial metabolism as glucose or glycine did not reduce signal in an analogous luxABCDE-modified Gram-positive pathogen, Staph. Xen29. The Strep. Xen20 cells produced light in a concentration-dependent manner, inversely related to the amount of glucose present. Taken together, our measures of microbial response could provide new information regarding the responsiveness of S. pyogenes metabolism to acute changes in its local environments and cellular health.
RESUMO
INTRODUCTION: Hands-on learning continues to serve as a positive mechanism for gaining interest and increasing recruitment in the health professions. This paper explores the Harrison School of Pharmacy's (HSOP) development and implementation of a week-long pharmacy camp designed to engage learners through active learning experiences to provide early exposure to the pharmacy profession. METHODS: The planning committee was formed in fall 2016, with the inaugural camp occurring summer 2017. A partnership with Auburn University Youth Programs allowed the committee to utilize existing university infrastructure and resources. Designed to expose campers to a variety of practice settings, the curriculum immersed participants in active learning experiences that allowed them to learn more about the clinical skills and knowledge needed for practice. To create diverse and learner-centric experiences, the planning committee recruited current second- and third-year student pharmacists to serve as counselors and peer instructors for all camp activities. RESULTS: Over two years, the camp hosted 40 campers representing nine states. Campers were predominantly female (65.3%), an average age of 16.8 years, and 16% were from diverse backgrounds. Camper feedback found overall satisfaction with the camp was high, with most indicating they attended camp to learn more about the pharmacy profession, specifically the school. As of spring 2019, 20% of total campers had applied and been accepted into the HSOP's Early Assurance Program. CONCLUSIONS: While early findings are good, the true value of the camp will be found over time as the committee explores if more students are choosing pharmacy as a career.