Strengthening Parent - Physician Communication: A Mixed Methods Study on Attuned Communication Training for Pediatric Residents.

Problem: High-quality communication improves patient satisfaction and clinical outcomes, yet formal communication training in residency is often minimal. Many studies on empathic communication show mixed results and are often hindered and skewed by brief study lengths, insufficiently and ambiguously defined concepts, and limited methods for objective measurements. Intervention: The FAN Curriculum is a unique communication curriculum, based on the conceptual frameworks of patient-centered communication, reflective practice, mindfulness, and attunement using the Facilitating Attuned Interactions (FAN) model. The first part of the FAN Curriculum was delivered as a 3-hour interactive workshop involving didactics, group discussion, and role play with pediatric residents. Residents then completed weekly self-reflections, a follow-up one-hour training to reinforce concepts, and five monthly mentor sessions, all emphasizing reflective practice. Context: This longitudinal, mixed-methods study examined the effects of the FAN Curriculum on residents' empathy levels and ability to communicate with parents in the clinical setting. The study was conducted at two urban, academic, medium-sized pediatric residency programs in Chicago between October 2016 and November 2017. First- and second-year pediatric residents whose continuity clinic site was located at their home institution participated. Residents received training in the use of the FAN Communication Tool using a delayed-start crossover study design. Impact: At five time points, residents and parents completed instruments validated for measuring physician empathy and mindfulness. Post-study interviews were conducted for one institution's residents and mentors and were evaluated using open and focused coding. Participants (n = 23) demonstrated a high degree of use of the FAN Communication Tool six months post-training and a significant rise in self-reported comfort with four of five FAN core processes. One parent-completed survey (Consultation and Relational Empathy, CARE) showed a statistically significant rise of 3.26% in resident relational empathy and collaboration after training (p = 0.02). In qualitative analysis of interviews, residents and mentors found the FAN Communication Tool beneficial, making clinic visits more efficient and collaborative. Both groups noted improvement in the residents' relationship-building skills; residents were able to use enhanced communication skills to better approach challenging encounters and work through parent concerns. Lessons Learned: Family-centered communication training can improve physician-perceived empathy and mindfulness. Effective communication for pediatric residents incorporates an empathic approach, and introduction to this formal curriculum supported their growth in connecting and engaging with children and parents. The FAN Curriculum may provide a useful method for improving resident communication skills with a positive impact on pediatricians' collaboration with patients and families.

Comparative phenotypic analysis of the major fungal pathogens Candida parapsilosis and Candida albicans.

Candida parapsilosis and Candida albicans are human fungal pathogens that belong to the CTG clade in the Saccharomycotina. In contrast to C. albicans, relatively little is known about the virulence properties of C. parapsilosis, a pathogen particularly associated with infections of premature neonates. We describe here the construction of C. parapsilosis strains carrying double allele deletions of 100 transcription factors, protein kinases and species-specific genes. Two independent deletions were constructed for each target gene. Growth in >40 conditions was tested, including carbon source, temperature, and the presence of antifungal drugs. The phenotypes were compared to C. albicans strains with deletions of orthologous transcription factors. We found that many phenotypes are shared between the two species, such as the role of Upc2 as a regulator of azole resistance, and of CAP1 in the oxidative stress response. Others are unique to one species. For example, Cph2 plays a role in the hypoxic response in C. parapsilosis but not in C. albicans. We found extensive divergence between the biofilm regulators of the two species. We identified seven transcription factors and one protein kinase that are required for biofilm development in C. parapsilosis. Only three (Efg1, Bcr1 and Ace2) have similar effects on C. albicans biofilms, whereas Cph2, Czf1, Gzf3 and Ume6 have major roles in C. parapsilosis only. Two transcription factors (Brg1 and Tec1) with well-characterized roles in biofilm formation in C. albicans do not have the same function in C. parapsilosis. We also compared the transcription profile of C. parapsilosis and C. albicans biofilms. Our analysis suggests the processes shared between the two species are predominantly metabolic, and that Cph2 and Bcr1 are major biofilm regulators in C. parapsilosis.

Mechanisms of Candida biofilm drug resistance.

Candida commonly adheres to implanted medical devices, growing as a resilient biofilm capable of withstanding extraordinarily high antifungal concentrations. As currently available antifungals have minimal activity against biofilms, new drugs to treat these recalcitrant infections are urgently needed. Recent investigations have begun to shed light on the mechanisms behind the profound resistance associated with the biofilm mode of growth. This resistance appears to be multifactorial, involving both mechanisms similar to conventional, planktonic antifungal resistance, such as increased efflux pump activity, as well as mechanisms specific to the biofilm lifestyle. A unique biofilm property is the production of an extracellular matrix. Two components of this material, ß-glucan and extracellular DNA, promote biofilm resistance to multiple antifungals. Biofilm formation also engages several stress response pathways that impair the activity of azole drugs. Resistance within a biofilm is often heterogeneous, with the development of a subpopulation of resistant persister cells. In this article we review the molecular mechanisms underlying Candida biofilm antifungal resistance and their relative contributions during various growth phases.

Preparation of Candida albicans Biofilms for Transmission Electron Microscopy.

Transmission Electron Microscopy is a form of microscopy that allows for imaging of distinct portions of an individual cell. For Candida albicans biofilms, it is often used to visualize the cell walls of fixed samples of yeast and hyphae. This protocol describes how to grow, harvest, and fix Candida albicans biofilms in preparation for Transmission Electron Microscopy.

Preparation of Candida albicans Biofilms Using an in vivo Rat Central Venous Catheter Model.

In vivo biofilms grown on medical devices are necessary to understand the interactions of the fungal biofilm and the host environment in which it is most commonly found. This protocol describes a way to grow Candida albicans biofilms on the interior lumen of central venous catheters surgically implanted into rats, which mimics quite well the clinical cases of biofilms found on human central venous catheters. These infected catheters can then be studied via a multitude of different experiments, including cell counting by plating, imaging the catheters under light or electron microscopy, or comparing the relative content of in vivo biofilms to in vitro biofilms and planktonic cultures. These biofilms also provide enough high quality RNA for transcriptional profiling.

A Candida biofilm-induced pathway for matrix glucan delivery: implications for drug resistance.

Extracellular polysaccharides are key constituents of the biofilm matrix of many microorganisms. One critical carbohydrate component of Candida albicans biofilms, ß-1,3 glucan, has been linked to biofilm protection from antifungal agents. In this study, we identify three glucan modification enzymes that function to deliver glucan from the cell to the extracellular matrix. These enzymes include two predicted glucan transferases and an exo-glucanase, encoded by BGL2, PHR1, and XOG1, respectively. We show that the enzymes are crucial for both delivery of ß-1,3 glucan to the biofilm matrix and for accumulation of mature matrix biomass. The enzymes do not appear to impact cell wall glucan content of biofilm cells, nor are they necessary for filamentation or biofilm formation. We demonstrate that mutants lacking these genes exhibit enhanced susceptibility to the commonly used antifungal, fluconazole, during biofilm growth only. Transcriptional analysis and biofilm phenotypes of strains with multiple mutations suggest that these enzymes act in a complementary fashion to distribute matrix downstream of the primary ß-1,3 glucan synthase encoded by FKS1. Furthermore, our observations suggest that this matrix delivery pathway works independently from the C. albicans ZAP1 matrix formation regulatory pathway. These glucan modification enzymes appear to play a biofilm-specific role in mediating the delivery and organization of mature biofilm matrix. We propose that the discovery of inhibitors for these enzymes would provide promising anti-biofilm therapeutics.

Portrait of Candida albicans adherence regulators.

Cell-substrate adherence is a fundamental property of microorganisms that enables them to exist in biofilms. Our study focuses on adherence of the fungal pathogen Candida albicans to one substrate, silicone, that is relevant to device-associated infection. We conducted a mutant screen with a quantitative flow-cell assay to identify thirty transcription factors that are required for adherence. We then combined nanoString gene expression profiling with functional analysis to elucidate relationships among these transcription factors, with two major goals: to extend our understanding of transcription factors previously known to govern adherence or biofilm formation, and to gain insight into the many transcription factors we identified that were relatively uncharacterized, particularly in the context of adherence or cell surface biogenesis. With regard to the first goal, we have discovered a role for biofilm regulator Bcr1 in adherence, and found that biofilm regulator Ace2 is a major functional target of chromatin remodeling factor Snf5. In addition, Bcr1 and Ace2 share several target genes, pointing to a new connection between them. With regard to the second goal, our findings reveal existence of a large regulatory network that connects eleven adherence regulators, the zinc-response regulator Zap1, and approximately one quarter of the predicted cell surface protein genes in this organism. This limited yet sensitive glimpse of mutant gene expression changes had thus defined one of the broadest cell surface regulatory networks in C. albicans.

Comparative analysis of Candida biofilm quantitation assays.

Candida grows on devices producing treatment resistant biofilms. A key tool for the study of biofilms includes an accurate assessment of viable cell growth. This study systematically tested seven techniques, among which the XTT assay provided the most reproducible, accurate, and efficient method for the quantitative estimation of C. albicans biofilms.