Integration of the Saline Process on Holistic Patient Care to Improve Student Understanding of Interprofessional Team Roles, Values, and Ethics

Description of the Problem: Healthcare practitioner students currently report feeling underprepared to provide holistic and spiritual care to their patients upon transitioning into practice, and there is currently little data on the efficacy of holistic care-focused interventions on interprofessional outcomes. The goal of this research was to assess the impact of an interprofessional training session on holistic care on student perceptions of interprofessional 1) roles/responsibilities and 2) values/ethics. The Innovation: A live, interactive interprofessional training session to address holistic patient care was implemented in fall of 2017. Students’ pre- and post-training perceptions of their confidence in study outcomes were assessed using a survey instrument. Critical Analysis: Significant positive changes were seen in students’ perceived ability to participate in team discussions and clarify misconceptions regarding their role in healthcare following the training. Students had high confidence in interacting ethically at pre-test and sustained that confidence. Next Steps: Live, interactive educational interventions with skills practice and group discussions can help to increase students’ awareness of team roles and responsibilities, as well as expand their understanding of the values and ethics within healthcare professions

RT-qPCR gene expression analysis in zebrafish : preanalytical precautions and use of expressed repetitive elements for normalization

Gene expression analysis is increasingly important in many fields of biological research. Understanding patterns of expressed genes is assumed to provide insight into complex regulatory networks and can lead to the identification of genes relevant to specific biological processes, including disease. Among different techniques, reverse transcription quantitative polymerase chain reaction (RT-qPCR) is currently regarded as the gold standard for targeted quantification of RNA gene expression, especially because of its high sensitivity, specificity, accuracy, and precision, and also because of its practical simplicity and processing speed. However, different critical factors can influence the outcome of RT-qPCR studies, including isolation of RNA, reverse transcription to cDNA, and data analysis. These factors need to be addressed in order to obtain biologically meaningful results. In this chapter, we describe how RT-qPCR can be used in a reliable way to successfully study differential gene expression in zebrafish. Hereby, we especially focus on how expressed repetitive elements can be employed as reference targets in zebrafish RT-qPCR studies and how they can further improve the quality of the data

Expressed repeat elements improve RT-qPCR normalization across a wide range of zebrafish gene expression studies

The selection and validation of stably expressed reference genes is a critical issue for proper RT-qPCR data normalization. In zebrafish expression studies, many commonly used reference genes are not generally applicable given their variability in expression levels under a variety of experimental conditions. Inappropriate use of these reference genes may lead to false interpretation of expression data and unreliable conclusions. In this study, we evaluated a novel normalization method in zebrafish using expressed repetitive elements (ERE) as reference targets, instead of specific protein coding mRNA targets. We assessed and compared the expression stability of a number of EREs to that of commonly used zebrafish reference genes in a diverse set of experimental conditions including a developmental time series, a set of different organs from adult fish and different treatments of zebrafish embryos including morpholino injections and administration of chemicals. Using geNorm and rank aggregation analysis we demonstrated that EREs have a higher overall expression stability compared to the commonly used reference genes. Moreover, we propose a limited set of ERE reference targets (loopern4, hatn10 and dna15ta1), that show stable expression throughout the wide range of experiments in this study, as strong candidates for inclusion as reference targets for qPCR normalization in future zebrafish expression studies. Our applied strategy to find and evaluate candidate expressed repeat elements for RT-qPCR data normalization has high potential to be used also for other species

Average expression stability of common reference genes and expressed repeat elements.

<p>Ranking of reference targets depending on their M-values calculated by geNorm. Reference targets with M-values below 0.5 and 0.2 are considered having a ‘high’ and ‘very high’ expression stability, respectively. EREs are indicated in black, commonly used reference mRNAs in grey.</p

Workflow to identify candidate expressed repeat elements.

<p>Workflow to identify candidate expressed repeat elements.</p

Reference target primer design and calculation of amplification efficiencies.

<p>(*)HUGO or repbase identifier.</p><p>Reference target primer design and calculation of amplification efficiencies.</p