Membrane-Permeant, Environment-Sensitive Dyes Generate Biosensors within Living Cells.

Dyes with environment-sensitive fluorescence have proven useful to study the spatiotemporal dynamics of protein activity in living cells. When attached to proteins, their fluorescence can reflect protein conformational changes, post-translational modifications, or protein interactions. However, the utility of such dye-protein conjugates has been limited because it is difficult to load them into cells. They usually must be introduced using techniques that perturb cell physiology, limit throughput, or generate fluorescent vesicles (e.g., electroporation, microinjection, or membrane transduction peptides). Here we circumvent these problems by modifying a proven, environment-sensitive biosensor fluorophore so that it can pass through cell membranes without staining intracellular compartments and can be attached to proteins within living cells using unnatural amino acid (UAA) mutagenesis. Reactive groups were incorporated for attachment to UAAs or small molecules (mero166, azide; mero167, alkyne; mero76, carboxylic acid). These dyes are bright and fluoresce at long wavelengths (reaching ε = 100 000 M-1 cm-1, ϕ = 0.24, with excitation 565 nm and emission 594 nm). The utility of mero166 was demonstrated by in-cell labeling of a UAA to generate a biosensor for the small GTPase Cdc42. In addition, conjugation of mero166 to a small molecule produced a membrane-permeable probe that reported the localization of the DNA methyltransferase G9a in cells. This approach provides a strategy to access biosensors for many targets and to more practically harness the varied environmental sensitivities of synthetic dyes.

Building Authentic Connection in the Patient-Physician Relationship.

INTRODUCTION/OBJECTIVES: Delivering optimal patient care is impacted by a physician's ability to build trusting relationships with patients. Identifying techniques for rapport building is important for promoting patient-physician collaboration and improved patient outcomes. This study sought to characterize the approaches highly skilled primary care physicians (PCPs) use to effectively connect with diverse patients. METHODS: Using an inductive thematic analysis approach, we analyzed semi-structured interview transcripts with 10 PCPs identified by leadership and/or colleagues for having exceptional patient communication skills. PCPs practiced in 3 diverse clinic settings: (1) academic medical center, (2) Veterans Affairs clinic, and (3) safety-net community clinic. RESULTS AND CONCLUSIONS: The thematic analysis yielded 5 themes that enable physicians to establish connections with patients: Respect for the Patient, Engaged Curiosity, Focused Listening, Mutual Participation, and Self-Awareness. Underlying all of these themes was a quality of authenticity, or a state of symmetry between one's internal experience and external words and actions. Adopting these communication techniques while allowing for adaptability in order to remain authentic in one's interactions with patients may facilitate improved connection and trust with patients. Encouraging physician authenticity in the patient-physician relationship supports a shift toward relationship-centered care. Additional medical education training is needed to facilitate authentic connection between physicians and patients.

Leadership development for orthopaedic trauma surgeons in Latin America: opportunities for and barriers to skill acquisition.

INTRODUCTION: There is growing interest in leadership courses for physicians. Few opportunities are available in global regions with limited resources. This study describes orthopaedic trauma surgeons' desired leadership skill acquisition, opportunities, and barriers to course participation in Latin America. METHODS: Latin American orthopaedic trauma surgeons from the Asociación de Cirujanos Traumatólogos de las Americas (ACTUAR) network were surveyed. This survey solicited and gauged the surgeons' level of interest in leadership topics and their relative importance utilizing a 5-point Likert-scale. Additionally, comparisons were calculated between middle-income countries (MICs) and high-income countries (HICs) to ascertain if needs were different between groups. The survey included demographic information, nationality, level of training, years in practice, leadership position, needs assessment, and perceived barriers for leadership educational opportunities. RESULTS: One hundred forty-four orthopaedic surgeons completed the survey, representing 18 countries across Latin America; 15 MICs and 3 HICs. Participants had more than 20 years in practice (49%) and held leadership positions (81%) in hospital settings (62%), national orthopaedic societies (45%), and/or clinical settings (40%). Sixty-three percent had never attended a leadership course due to lack of opportunities/invitations (69%), difficulty missing work (24%), and costs (21%). Ninety-seven percent expressed interest in attending a leadership course. No difference in needs was determined between respondents from MICs and HICs. Professional Ethics, Crisis Management/Organizational Change Management, and High Performing Team-Building were identified as the most important leadership topics. CONCLUSION: Orthopaedic surgeons in Latin America demonstrate an interest in acquiring additional leadership skills but have few opportunities. Identifying interests, knowledge gaps, and core competencies can guide the development of such opportunities.