Engagement Across Professions: A Mixed Methods Study of Debriefing After Interprofessional Team Training.

INTRODUCTION: Simulation is an ideal tool for interprofessional (IP) team training. Debriefing after simulation is key to IP learning, although engagement and participation may be adversely influenced by cultural and hierarchical barriers. This mixed-methods study explored factors influencing learner engagement and participation in IP debriefing and the experience of "silent but apparently engaged" participants. METHODS: Semistructured profession-specific focus groups were conducted with participants from a weekly IP pediatric simulation program. Focus groups were recorded, transcribed, and thematically analyzed. Eligible participants were assigned to "silent" or "verbal" groups according to observed behavior and received a questionnaire. Participants' self-rated engagement scores were compared using a t test. RESULTS: Thirty-six of 81 eligible participants were included, 13 completed a questionnaire, and 23 (8 physicians, 10 nursing staff, 4 pharmacists, 1 respiratory therapist) participated in 13 focus groups. Twenty-two subthemes were grouped into 6 themes: psychological safety, realism, distractors, stress, group characteristics, and facilitator behavior, with differences in perspective according to profession. Of the 36 respondents, 18 were "silent" and 18 "verbal." Self-rated engagement scores differed between groups (3.65 vs. 4.17, P = 0.06); however, "silent" participants described themselves as engaged. CONCLUSIONS: Themes identified that influenced learner engagement in debriefing included aspects of prebriefing and the simulation. Some aligned with general simulation best practices, such as psychological safety, prebriefing, and facilitator behavior. Findings unique to IP simulation included importance of realism to nonphysician professions, protecting time for training, group composition, and direct probing by cofacilitators to decrease physician bias and emphasize IP contributions. Silent participants reported engagement.

Spiritual care in Australian general practice nursing: An interpretive descriptive study.

Spiritual care as an aspect of holistic or person-centered care has been well documented. Studies on spirituality and spiritual care in nursing have taken place in various countries and contexts. Studies about spiritual care from the nurse perspective in the primary healthcare setting of Australian General Practice are not evident. Using an interpretive description study design, data about views on spirituality, spiritual care experiences, and descriptions about any spiritual care provided were collected from eight practice nurses. Four themes were drawn from the findings: the personal importance of spirituality and spiritual care to nurses and patients, spiritual care as an aspect of person-centered care, spiritual care practices, and barriers and enablers to addressing spiritual needs. Participants identified that practice nurses require improved education about spirituality to adequately address spiritual needs as expressed by patients, as well as the tools to be able to provide spiritual care. They also identified that documentation templates used in general practice should include prompts to address spiritual and/or religious needs.

Factors associated with interprofessional engagement in debriefing following pediatric simulation-based team training.

Simulation offers a high fidelity modality to deliver and study team-based interprofessional education. Debriefing the following simulated scenarios is a critical component of this training. Little data exist to inform best practices to optimize interprofessional engagement during debriefing. This pilot study analyzed interprofessional debriefing events following 20 pediatric simulation-based team trainings to identify associations between modifiable factors and learner engagement. Reviewers observed a total of 236 learners, using a previously published tool to assess learner engagement. Data related to the scenario, debriefing, learners, and facilitators were collected. Spearman's correlation was used to analyze the association between factors of interest and average learner engagement scores for each debriefing event. Mean engagement did not differ between physicians and nurses, but was lower for other professionals. Average learner engagement was inversely related to learner group size, but not to the proportion of learners in each profession. Oral participation differed significantly between professions for both learners and co-facilitators, with physicians speaking more in both groups. Students of all professions had lower engagement and spoke less frequently. This study identifies several modifiable factors, including total group size, learner level, and facilitator behavior that were associated with interprofessional engagement during debriefing following simulation-based team training.

Single-cell cloning enables the selection of more productive Drosophila melanogaster S2 cells for recombinant protein expression.

The generation of monoclonal cell lines is an important early process development step for recombinant protein production. Although single-cell cloning is an established method in mammalian cell lines, straightforward protocols are not yet available for insect cells. We describe a new method for the generation of monoclonal insect cells without using fetal bovine serum and/or feeder cells pretreated by irradiation or exposure to mitomycin. Highly productive clones of Drosophila melanogaster S2 cells were prepared in a two-step procedure, comprising the establishment of a polyclonal population and subsequent single cell isolation by limiting dilution. Necessary growth factors were provided by co-cultivation of single transformants with untransfected feeder cells, which were later removed by antibiotic selection. Enhanced expression of EGFP and two target peptides was confirmed by flow cytometry and dot/western blotting. Highly productive clones were stable, showed a uniform expression profile and typically a sixfold to tenfold increase in cell-specific productivity.

A high-throughput expression screening platform to optimize the production of antimicrobial peptides.

BACKGROUND: Antimicrobial peptides (AMPs) are promising candidates for the development of novel antibiotics, but it is difficult to produce sufficient quantities for preclinical and clinical studies due to their toxicity towards microbial expression hosts. To avoid laborious trial-and-error testing for the identification of suitable expression constructs, we have developed a small-scale expression screening platform based on a combinatorial plasmid library. RESULTS: The combinatorial library is based on the Golden Gate cloning system. In each reaction, six donor plasmids (each containing one component: a promoter, fusion partner 1, fusion partner 2, protease cleavage site, gene of interest, or transcriptional terminator) were combined with one acceptor plasmid to yield the final expression construct. As a proof of concept, screening was carried out in Escherichia coli and Pichia pastoris to study the expression of three different model AMPs with challenging characteristics, such as host toxicity or multiple disulfide bonds. The corresponding genes were successfully cloned in 27 E. coli and 18 P. pastoris expression plasmids, each in a one-step Golden Gate reaction. After transformation, small-scale expression screening in microtiter plates was followed by AMP quantification using a His6 tag-specific ELISA. Depending on the plasmid features and the expression host, the protein yields differed by more than an order of magnitude. This allowed the identification of high producers suitable for larger-scale protein expression. CONCLUSIONS: The optimization of recombinant protein production is best achieved from first principles by initially optimizing the genetic construct. The unrestricted combination of multiple plasmid features yields a comprehensive library of expression strains that can be screened for optimal productivity. The availability of such a platform could benefit all laboratories working in the field of recombinant protein expression.

The potential of the Galleria mellonella innate immune system is maximized by the co-presentation of diverse antimicrobial peptides.

Antimicrobial peptides (AMPs) are ubiquitous components of the insect innate immune system. The model insect Galleria mellonella has at least 18 AMPs, some of which are still uncharacterized in terms of antimicrobial activity. To determine why G. mellonella secretes a repertoire of distinct AMPs following an immune challenge, we selected three different AMPs: cecropin A (CecA), gallerimycin and cobatoxin. We found that cobatoxin was active against Micrococcus luteus at a minimum inhibitory concentration (MIC) of 120 µm, but at 60 µm when co-presented with 4 µm CecA. In contrast, the MIC of gallerimycin presented alone was 60 µm and the co-presentation of CecA did not affect this value. Cobatoxin and gallerimycin were both inactive against Escherichia coli at physiological concentrations, however gallerimycin could potentiate the sublethal dose of CecA (0.25 µm) at a concentration of 30 µm resulting in 100% lethality. The ability of gallerimycin to potentiate the CecA was investigated by flow cytometry, revealing that 30 µm gallerimycin sensitized E. coli cells by inducing membrane depolarization, which intensified the otherwise negligible effects of 0.25 µm CecA. We therefore conclude that G. mellonella maximizes the potential of its innate immune response by the co-presentation of different AMPs that become more effective at lower concentrations when presented simultaneously.

A soluble fragment of the tumor antigen BCL2-associated athanogene 6 (BAG-6) is essential and sufficient for inhibition of NKp30 receptor-dependent cytotoxicity of natural killer cells.

Immunosurveillance of tumor cells depends on NKp30, a major activating receptor of human natural killer (NK) cells. The human BCL2-associated athanogene 6 (BAG-6, also known as BAT3; 1126 amino acids) is a cellular ligand of NKp30. To date, little is known about the molecular details of this receptor ligand system. Within the current study, we have located the binding site of NKp30 to a sequence stretch of 250 amino acids in the C-terminal region of BAG-6 (BAG-6(686-936)). BAG-6(686-936) forms a noncovalent dimer of 57-59 kDa, which is sufficient for high affinity interaction with NKp30 (KD < 100 nM). As our most important finding, BAG-6(686-936) inhibits NKp30-dependent signaling, interferon-γ release, and degranulation of NK cells in the presence of malignantly transformed target cells. Based on these data, we show for the first time that BAG-6(686-936) comprises a subdomain of BAG-6, which is sufficient for receptor docking and inhibition of NKp30-dependent NK cell cytotoxicity as part of a tumor immune escape mechanism. These molecular insights provide an access point to restore tumor immunosurveillance by NK cells and to increase the efficacy of cellular therapies.

Organ identity and environmental conditions determine the effectiveness of nonhost resistance in the interaction between Arabidopsis thaliana and Magnaporthe oryzae.

Mechanisms leading to nonhost resistance of plants against nonadapted pathogens are thought to have great potential for the future management of agriculturally important diseases. In this article, we report an investigation of nonhost resistance motivated by the advantages of studying an interaction between two model organisms, namely Arabidopsis thaliana and Magnaporthe oryzae. During the course of our studies, however, we discovered an unexpected plasticity in the responses of Arabidopsis against this ostensibly nonhost pathogen. Thus, we elucidated that certain experimental conditions, such as the growth of plants under long days at constantly high humidity and the use of high inoculum concentrations of M. oryzae conidia, forced the interaction in leaves of some Arabidopsis ecotypes towards increased compatibility. However, sporulation was never observed. Furthermore, we observed that roots were generally susceptible to M. oryzae, whereas leaves, stems and hypocotyls were not infected. It must be concluded, therefore, that Arabidopsis roots lack an effective defence repertoire against M. oryzae, whereas its leaves possess such nonhost defence mechanisms. In summary, our findings point to organ-specific determinants and environmental conditions influencing the effectiveness of nonhost resistance in plants.