Constitutive androstane receptor (CAR, NR1I3) belongs to the nuclear receptor family of transcription factors and acts as a chemical sensor of drugs and endogenous compounds. The ligand-binding preferences of CAR are diverse, and more importantly, there are significant species differences in ligand specificity. Here, we show that while certain residues are critical for the basal activity of mouse CAR (mCAR) and/or affect the binding of all tested ligands, mutation of some ligand-binding pocket (LBP) residues (e.g., F171 and Y336) paradoxically decreased the activity of a specific ligand while increasing that of others. Comparisons to previously reported human CAR (hCAR) residues indicated that the function of key CAR residues (e.g., N175, L253) is dramatically different between species. The docking results provide some mechanistic rationale for the ability of 17α-ethinyl-3,17β-estradiol (EE2) to both activate mCAR and repress hCAR.

Ligand specificity of constitutive androstane receptor as probed by induced-fit docking and mutagenesis.

Objective: To assess student learning and perceived task value of physical pharmacy wet laboratory sessions conducted at two separate campuses within a school of pharmacy. Methods: Anonymous archival course evaluation and assessment data from 130 students enrolled in a pharmaceutics course were used. The evaluation surveyed students about the task value of five wet laboratory sessions, and assessment data were of preand post lab knowledge. Campuses differed in lecture delivery (in-person vs. video-conferencing), but labs were the same. Results: Most students felt that the quality and organization of the lab sessions were good, that the lab sessions allowed them to better understand pharmaceutics concepts, and that the time spent in lab was worthwhile. Most students also indicated that they preferred wet labs over virtual labs or no labs. The proportion of students achieving mastery on knowledge assessments increased significantly from prelab to postlab. No meaningful differences were found between locations. Conclusion: Wet laboratory exercises are a useful supplement for learning physical pharmacy concepts.

Assessment of Students’ Learning and Perceptions of Task Value of a Physical Pharmacy Laboratory Course

In the world of higher education, one of the struggles instructors face in the classroom is engaging students in the material. A second discussion in higher education pedagogy is how to weigh conte...

Utilizing Saccharomyces in the classroom: a versatile organism for teaching and learning

Objectives. Nuclear receptors (NRs) are ligand-activated transcription factors that regulate development and cellular differentiation, endocrine functions, and metabolism and transport of nutrients and drugs. Understanding of NR action is therefore essential for students of pharmacy and biology. Currently, there are no convenient laboratory exercises that clearly demonstrate the components and mechanisms of NR action. Design. Employing the yeast two-hybrid method, we developed a simple hands-on exercise that demonstrated ligand binding, dose responses, and consequences of ligand binding that are critical for NR function. Assessment. The exercise uses inexpensive, non-radioactive reagents and standard laboratory equipment. The results are directly visible, easy to interpret, and reproducible. The whole exercise can be conducted in a single day. In a real life test, a majority of the students who participated reported that their understanding of NR action had increased. Conclusions. This simple exercise can...

Teaching the Basics of Nuclear Receptor Action: A Simple Laboratory Exercise Using the Yeast Two-Hybrid Method

Objectives. Nuclear receptors (NRs) are ligand-activated transcription factors that regulate development and cellular differentiation, endocrine functions, and metabolism and transport of nutrients and drugs. Understanding of NR action is therefore essential for students of pharmacy and biology. Currently, there are no convenient laboratory exercises that clearly demonstrate the components and mechanisms of NR action. Design. Employing the yeast two-hybrid method, we developed a simple hands-on exercise that demonstrated ligand binding, dose responses, and consequences of ligand binding that are critical for NR function. Assessment. The exercise uses inexpensive, non-radioactive reagents and standard laboratory equipment. The results are directly visible, easy to interpret, and reproducible. The whole exercise can be conducted in a single day. In a real life test, a majority of the students who participated reported that their understanding of NR action had increased. Conclusions. This simple exercise can be applied to other NRs and modified to demonstrate pharmacological concepts including competitive antagonism, partial agonism, and structure-activity relationship.

http://archive.ajpe.org/aj6902/aj690226/aj690226.pdf

Silvia Petersen

Papers

Teaching the Basics of Nuclear Receptor Action: A Simple Laboratory Exercise Using the Yeast Two-Hybrid Method

INSTRUCTIONAL DESIGN AND ASSESSMENT Teaching the Basics of Nuclear Receptor Action: A Simple Laboratory Exercise Using the Yeast Two-Hybrid Method