Past, present and future of respiratory research: A survey of Canadian health care professionals.

BACKGROUND: The Canadian Respiratory Health Professionals (CRHP) is the multidisciplinary health care professional group of the Canadian Lung Association. Although the CRHP has a growing number of highly qualified researchers, the landscape of their research in Canada has not been described. OBJECTIVES: To describe the level of respiratory research engagement; identify barriers and facilitators to research engagement; describe the experience and interest in developing research skills; and identify priority areas of future respiratory research among health care professionals. METHODS: An online survey of CRHP members was used to collect demographic information; barriers and facilitators to conducting research; future directions in respiratory research; and research funding and mentorship. Experience with and interest in 'upskilling' research skills were also evaluated. RESULTS: A total of 119 surveys were completed (22% response rate), of which 69 (58%) respondents were engaged in respiratory research. Reasons for not being involved in respiratory research were lack of mentorship, support and funding. The top research areas were chronic obstructive pulmonary disease (74%) and asthma (41%). The top facilitators for research engagement were amount of funding (29%) and mentorship (28%). Respondents in research positions rated their experience in research skills as high; those in nonresearch positions as low. However, both groups expressed interest in improving their research skills. CONCLUSIONS: Areas of development, such as research skills, greater funding opportunities and mentorship to increase the research capacity of health care professionals in respiratory health were identified. Health professional researchers have an important role in the national respiratory research strategy to increase interdisciplinary engagement and build collaborative teams.

Lactone-bound structures of cyclohexanone monooxygenase provide insight into the stereochemistry of catalysis.

The Baeyer-Villiger monooxygenases (BVMOs) are microbial enzymes that catalyze the synthetically useful Baeyer-Villiger oxidation reaction. The available BVMO crystal structures all lack a substrate or product bound in a position that would determine the substrate specificity and stereospecificity of the enzyme. Here, we report two crystal structures of cyclohexanone monooxygenase (CHMO) with its product, ε-caprolactone, bound: the CHMO(Tight) and CHMO(Loose) structures. The CHMO(Tight) structure represents the enzyme state in which substrate acceptance and stereospecificity is determined, providing a foundation for engineering BVMOs with altered substrate spectra and/or stereospecificity. The CHMO(Loose) structure is the first structure where the product is solvent accessible. This structure represents the enzyme state upon binding and release of the substrate and product. In addition, the role of the invariant Arg329 in chaperoning the substrate/product during the catalytic cycle is highlighted. Overall, these data provide a structural framework for the engineering of BVMOs with altered substrate spectra and/or stereospecificity.

The substrate-bound crystal structure of a Baeyer-Villiger monooxygenase exhibits a Criegee-like conformation.

The Baeyer-Villiger monooxygenases (BVMOs) are a family of bacterial flavoproteins that catalyze the synthetically useful Baeyer-Villiger oxidation reaction. This involves the conversion of ketones into esters or cyclic ketones into lactones by introducing an oxygen atom adjacent to the carbonyl group. The BVMOs offer exquisite regio- and enantiospecificity while acting on a wide range of substrates. They use only NADPH and oxygen as cosubstrates, and produce only NADP(+) and water as byproducts, making them environmentally attractive for industrial purposes. Here, we report the first crystal structure of a BVMO, cyclohexanone monooxygenase (CHMO) from Rhodococcus sp. HI-31 in complex with its substrate, cyclohexanone, as well as NADP(+) and FAD, to 2.4 Å resolution. This structure shows a drastic rotation of the NADP(+) cofactor in comparison to previously reported NADP(+)-bound structures, as the nicotinamide moiety is no longer positioned above the flavin ring. Instead, the substrate, cyclohexanone, is found at this location, in an appropriate position for the formation of the Criegee intermediate. The rotation of NADP(+) permits the substrate to gain access to the reactive flavin peroxyanion intermediate while preventing it from diffusing out of the active site. The structure thus reveals the conformation of the enzyme during the key catalytic step. CHMO is proposed to undergo a series of conformational changes to gradually move the substrate from the solvent, via binding in a solvent excluded pocket that dictates the enzyme's chemospecificity, to a location above the flavin-peroxide adduct where catalysis occurs.

Teaching professionalism through virtual means.

OBJECTIVES: Virtual patients are used across a variety of clinical disciplines for both teaching and assessment, but are they an appropriate environment in which to develop professional skills? This study aimed to evaluate students' perceived effectiveness of an online interactive virtual patient developed to augment a personal professional development curriculum, and to identify factors that would maximise the associated educational benefits. METHODS: Student focus group discussions were conducted to explore students' views on the usefulness and acceptability of the virtual patient as an educational tool to teach professionalism, and to identify factors for improvement. A thematic content analysis was used to capture content and synthesise the range of opinions expressed. RESULTS: Overall there was a positive response to the virtual patient. The students recognised the need to teach and assess professionalism throughout their curriculum, and viewed the virtual patient as a potentially engaging and valuable addition to their curriculum. We identified factors for improvement to guide the development of future virtual patients. CONCLUSION: It is possible to improve approaches to teaching and learning professionalism by exploring students' views on innovative teaching developments designed to augment personal professional development curricula.