Emotional regulation and suicidal ideation-Mediating roles of perceived social support and avoidant coping.

Introduction: Recent research has uncovered a wide prevalence variation of suicidal ideation in university students ranging from 9.7% to 58.3%. India has witnessed a 4.5% increase in suicide rates in the year 2021. The interplay between cognitive reappraisal of a stressful situation, suppression of emotional expression, and coping strategies for suicidal ideation of Indian University students is yet to be explored. We aim to determine whether suicidal ideation would differ across different types of family units, and to predict the extent to which perceived social support and avoidant coping could mediate the relation between emotion regulation processes and suicidal ideation. Methods: Two hundred randomly selected University students (Mean age = 19.9, SD = 1.43) participated. Kruskal-Wallis, Pearson's product-moment correlation, and GLM mediation model were computed. Results and discussion: Lifetime suicidal ideation significantly differed between those who stay alone and those who live in a nuclear family (p < 0.01), and also those who stay in a joint family (p < 0.05). Cognitive reappraisal predicted a reduction in suicidal ideation mediated by perceived social support (B = -0.06, p < 0.05) and avoidant coping (B = -0.07, p < 0.05). Whereas, expressive suppression predicted induced levels of suicidal ideation through perceived social support (B = 0.05, p < 0.05), and avoidant coping (B = 0.06, p < 0.05) as mediators. Conclusion: Though our sample size restricts the generalization, our findings implied the importance of regular psychological consultation regarding the efficacy of the said coping processes in dealing with suicidal ideation.

Insomnia among community members in Florida: Associations with demographics, health conditions, and social support.

Objectives: To identify associations between demographics, social determinants of health, health conditions, and reported history of insomnia. A cross-sectional study including 11,960 adult community members recruited through HealthStreet, a community outreach program at University of Florida. Methods: Health assessments were conducted via interviews. Participants reported their demographic background, level of social support, history of health conditions, and insomnia. Logistic regression was used to understand associations between risk factors and history of insomnia. Results: The prevalence of self-reported insomnia was 27.3%. Adults aged ≥ 65 years (OR = 1.16) and women (OR = 1.18) reported higher rates of insomnia than their counterparts. Black/African American individuals reported lower rates of insomnia (OR = 0.72) than White individuals. Individuals with food insecurity (OR = 1.53), a military history (OR = 1.30), lower social support (OR = 1.24), living alone (OR = 1.14), anxiety (OR = 2.33), cardiometabolic disease (OR = 1.58), and attention-deficit hyperactivity disorder (ADHD) (OR = 1.44) were significantly more likely to endorse insomnia compared with their counterparts. Depression (OR = 2.57) had the strongest association with insomnia. Conclusions: This study provides evidence regarding who is at greater risk for insomnia among a large community-based sample. Our findings highlight the importance of screening for insomnia, particularly among patients who experience food insecurity, are military veterans, have anxiety, depression, ADHD, or cardiometabolic disease, as well as those who live alone or have lower levels of social support. Future public health campaigns should provide education on insomnia symptoms, treatments, and evidenced-based sleep-promotion strategies.

Machine learning functional impairment classification with electronic health record data.

BACKGROUND: Poor functional status is a key marker of morbidity, yet is not routinely captured in clinical encounters. We developed and evaluated the accuracy of a machine learning algorithm that leveraged electronic health record (EHR) data to provide a scalable process for identification of functional impairment. METHODS: We identified a cohort of patients with an electronically captured screening measure of functional status (Older Americans Resources and Services ADL/IADL) between 2018 and 2020 (N = 6484). Patients were classified using unsupervised learning K means and t-distributed Stochastic Neighbor Embedding into normal function (NF), mild to moderate functional impairment (MFI), and severe functional impairment (SFI) states. Using 11 EHR clinical variable domains (832 variable input features), we trained an Extreme Gradient Boosting supervised machine learning algorithm to distinguish functional status states, and measured prediction accuracies. Data were randomly split into training (80%) and test (20%) sets. The SHapley Additive Explanations (SHAP) feature importance analysis was used to list the EHR features in rank order of their contribution to the outcome. RESULTS: Median age was 75.3 years, 62% female, 60% White. Patients were classified as 53% NF (n = 3453), 30% MFI (n = 1947), and 17% SFI (n = 1084). Summary of model performance for identifying functional status state (NF, MFI, SFI) was AUROC (area under the receiving operating characteristic curve) 0.92, 0.89, and 0.87, respectively. Age, falls, hospitalization, home health use, labs (e.g., albumin), comorbidities (e.g., dementia, heart failure, chronic kidney disease, chronic pain), and social determinants of health (e.g., alcohol use) were highly ranked features in predicting functional status states. CONCLUSION: A machine learning algorithm run on EHR clinical data has potential utility for differentiating functional status in the clinical setting. Through further validation and refinement, such algorithms can complement traditional screening methods and result in a population-based strategy for identifying patients with poor functional status who need additional health resources.

Provider Interaction With an Electronic Health Record Notification to Identify Eligible Patients for a Cluster Randomized Trial of Advance Care Planning in Primary Care: Secondary Analysis.

BACKGROUND: Advance care planning (ACP) improves patient-provider communication and aligns care to patient values, preferences, and goals. Within a multisite Meta-network Learning and Research Center ACP study, one health system deployed an electronic health record (EHR) notification and algorithm to alert providers about patients potentially appropriate for ACP and the clinical study. OBJECTIVE: The aim of the study is to describe the implementation and usage of an EHR notification for referring patients to an ACP study, evaluate the association of notifications with study referrals and engagement in ACP, and assess provider interactions with and perspectives on the notifications. METHODS: A secondary analysis assessed provider usage and their response to the notification (eg, acknowledge, dismiss, or engage patient in ACP conversation and refer patient to the clinical study). We evaluated all patients identified by the EHR algorithm during the Meta-network Learning and Research Center ACP study. Descriptive statistics compared patients referred to the study to those who were not referred to the study. Health care utilization, hospice referrals, and mortality as well as documentation and billing for ACP and related legal documents are reported. We evaluated associations between notifications with provider actions (ie, referral to study, ACP not documentation, and ACP billing). Provider free-text comments in the notifications were summarized qualitatively. Providers were surveyed on their satisfaction with the notification. RESULTS: Among the 2877 patients identified by the EHR algorithm over 20 months, 17,047 unique notifications were presented to 45 providers in 6 clinics, who then referred 290 (10%) patients. Providers had a median of 269 (IQR 65-552) total notifications, and patients had a median of 4 (IQR 2-8). Patients with more (over 5) notifications were less likely to be referred to the study than those with fewer notifications (57/1092, 5.2% vs 233/1785, 13.1%; P<.001). The most common free-text comment on the notification was lack of time. Providers who referred patients to the study were more likely to document ACP and submit ACP billing codes (P<.001). In the survey, 11 providers would recommend the notification (n=7, 64%); however, the notification impacted clinical workflow (n=9, 82%) and was difficult to navigate (n=6, 55%). CONCLUSIONS: An EHR notification can be implemented to remind providers to both perform ACP conversations and refer patients to a clinical study. There were diminishing returns after the fifth EHR notification where additional notifications did not lead to more trial referrals, ACP documentation, or ACP billing. Creation and optimization of EHR notifications for study referrals and ACP should consider the provider user, their workflow, and alert fatigue to improve implementation and adoption. TRIAL REGISTRATION: ClinicalTrials.gov NCT03577002; https://clinicaltrials.gov/ct2/show/NCT03577002.

Mistranslation of the genetic code by a new family of bacterial transfer RNAs.

The correct coupling of amino acids with transfer RNAs (tRNAs) is vital for translating genetic information into functional proteins. Errors during this process lead to mistranslation, where a codon is translated using the wrong amino acid. While unregulated and prolonged mistranslation is often toxic, growing evidence suggests that organisms, from bacteria to humans, can induce and use mistranslation as a mechanism to overcome unfavorable environmental conditions. Most known cases of mistranslation are caused by translation factors with poor substrate specificity or when substrate discrimination is sensitive to molecular changes such as mutations or posttranslational modifications. Here we report two novel families of tRNAs, encoded by bacteria from the Streptomyces and Kitasatospora genera, that adopted dual identities by integrating the anticodons AUU (for Asn) or AGU (for Thr) into the structure of a distinct proline tRNA. These tRNAs are typically encoded next to a full-length or truncated version of a distinct isoform of bacterial-type prolyl-tRNA synthetase. Using two protein reporters, we showed that these tRNAs translate asparagine and threonine codons with proline. Moreover, when expressed in Escherichia coli, the tRNAs cause varying growth defects due to global Asn-to-Pro and Thr-to-Pro mutations. Yet, proteome-wide substitutions of Asn with Pro induced by tRNA expression increased cell tolerance to the antibiotic carbenicillin, indicating that Pro mistranslation can be beneficial under certain conditions. Collectively, our results significantly expand the catalog of organisms known to possess dedicated mistranslation machinery and support the concept that mistranslation is a mechanism for cellular resiliency against environmental stress.

Using Design Thinking to Promote Goals of Care Conversations With Seriously Ill Patients.

BACKGROUND: Goals of care (GOC) conversations can improve serious illness outcomes such as pain and symptom management and patient satisfaction. PROBLEM: However, we recognized that very few Duke Health patients who died had a GOC conversation documented in the designated electronic health record (EHR) tab. Therefore, in 2020, we set a target that all Duke Health patients who died should have had a GOC conversation documented in a designated EHR tab in the last 6 months of life. INTERVENTION: In developing a strategy to promote GOC conversations, we used two interwoven approaches. The first was RE-AIM, a model for designing, reporting and evaluating health behavior research. The second was less of a model than a way of approaching problems, known as "design thinking." OUTCOMES: We employed both of these approaches in a system-wide effort that achieved a 50% prevalence of GOC conversations in the last 6 months of life. KEY MESSAGE: In combination, simple interventions can have a significant impact on behavior change in an academic health system. LESSONS LEARNED: We found that design thinking techniques offered a useful bridge between RE-AIM strategy and clinical.

RNA-controlled nucleocytoplasmic shuttling of mRNA decay factors regulates mRNA synthesis and a novel mRNA decay pathway.

mRNA level is controlled by factors that mediate both mRNA synthesis and decay, including the 5' to 3' exonuclease Xrn1. Here we show that nucleocytoplasmic shuttling of several yeast mRNA decay factors plays a key role in determining both mRNA synthesis and decay. Shuttling is regulated by RNA-controlled binding of the karyopherin Kap120 to two nuclear localization sequences (NLSs) in Xrn1, location of one of which is conserved from yeast to human. The decaying RNA binds and masks NLS1, establishing a link between mRNA decay and Xrn1 shuttling. Preventing Xrn1 import, either by deleting KAP120 or mutating the two Xrn1 NLSs, compromises transcription and, unexpectedly, also cytoplasmic decay, uncovering a cytoplasmic decay pathway that initiates in the nucleus. Most mRNAs are degraded by both pathways - the ratio between them represents a full spectrum. Importantly, Xrn1 shuttling is required for proper responses to environmental changes, e.g., fluctuating temperatures, involving proper changes in mRNA abundance and in cell proliferation rate.

Directed Evolution of Methanomethylophilus alvus Pyrrolysyl-tRNA Synthetase Generates a Hyperactive and Highly Selective Variant.

Pyrrolysyl-tRNA synthetase (PylRS) is frequently used for site-specific incorporation of noncanonical amino acids (ncAAs) into proteins. Recently, the active site of Methanomethylophilus alvus PylRS (MaPylRS) has been rationally engineered to expand its substrate compatibility, enabling the incorporation of difficult ncAAs. However, mutations beyond the active site that enhance the enzymatic properties of MaPylRS have not been reported. We utilized phage-assisted non-continuous evolution (PANCE) to evolve MaPylRS to efficiently incorporate N ε-Boc-l-lysine (BocK). Directed evolution yielded several mutations outside of the active site that greatly improve the activity of the enzyme. We combined the most effective mutations to generate a new PylRS variant (PylRSopt) that is highly active and selective towards several lysine and phenylalanine derivatives. The mutations in PylRSopt can be used to enhance previously engineered PylRS constructs such as MaPylRSN166S, and PylRSopt is compatible in applications requiring dual ncAA incorporation and substantially improves the yield of these target proteins.

Robust and annotation-free analysis of alternative splicing across diverse cell types in mice.

Although alternative splicing is a fundamental and pervasive aspect of gene expression in higher eukaryotes, it is often omitted from single-cell studies due to quantification challenges inherent to commonly used short-read sequencing technologies. Here, we undertake the analysis of alternative splicing across numerous diverse murine cell types from two large-scale single-cell datasets-the Tabula Muris and BRAIN Initiative Cell Census Network-while accounting for understudied technical artifacts and unannotated events. We find strong and general cell-type-specific alternative splicing, complementary to total gene expression but of similar discriminatory value, and identify a large volume of novel splicing events. We specifically highlight splicing variation across different cell types in primary motor cortex neurons, bone marrow B cells, and various epithelial cells, and we show that the implicated transcripts include many genes which do not display total expression differences. To elucidate the regulation of alternative splicing, we build a custom predictive model based on splicing factor activity, recovering several known interactions while generating new hypotheses, including potential regulatory roles for novel alternative splicing events in critical genes like Khdrbs3 and Rbfox1. We make our results available using public interactive browsers to spur further exploration by the community.

Cells are the basic building blocks of all living things. There are numerous types of cells, and each cell has its own machinery to fulfill a specialised role. Despite their different purposes, most cells contain the same instructions, stored as DNA, on how to assemble the proteins needed to perform their intended functions. Cell types often vary in the frequency that each gene is read, leading to different quantities of proteins produced. Moreover, a process known as alternative splicing enables cells to build multiple proteins from the same gene. It works by joining fragments of a gene's code in various combinations. The resulting RNA sequences are molecular templates that cells use to assemble proteins. Analysing these RNA sequences reveals which genes are switched on in different tissues of the body, and what proteins are being made. However, despite recent advancements, alternative splicing is rarely studied in single cells because of some sizeable technical challenges. Benegas, Fischer and Song developed a computational toolkit designed to handle the unique challenges of analysing alternative splicing events in single cells. The analysis pipeline, called scQuint, was tested on two large datasets that capture cell-to-cell differences in the brain and other tissues of mice. Nearly all the cell types studied exhibited clear differences in alternative splicing, such that cell types could be distinguished based on their splicing profiles. Intriguing patterns of splicing were highlighted in some immune cells and certain types of neurons. Across cell types, the genes with unique splicing patterns were often not the same as those with unique activity patterns, indicating that gene expression and alternative splicing are two complementary processes. New types of alternative splicing events were also identified. Benegas et al. also developed a statistical model to probe the roles of splicing regulators in different cell types. In summary, the scQuint toolkit overcomes critical technical challenges typically encountered when analysing alternative splicing in single cells. It also reveals new insights about mechanisms of alternative splicing. The results are open access, made available using public interactive browsers, which should spur on other researchers to interrogate how alternative splicing differs in single cells.

Association of an Advance Care Planning Video and Communication Intervention With Documentation of Advance Care Planning Among Older Adults: A Nonrandomized Controlled Trial.

Importance: COVID-19 has disproportionately killed older adults and racial and ethnic minority individuals, raising questions about the relevance of advance care planning (ACP) in this population. Video decision aids and communication skills training offer scalable delivery models. Objective: To assess whether ACP video decision aids and a clinician communication intervention improved the rate of ACP documentation during an evolving pandemic, with a focus on African American and Hispanic patients. Design, Setting, and Participants: The Advance Care Planning: Communicating With Outpatients for Vital Informed Decisions trial was a pre-post, open-cohort nonrandomized controlled trial that compared ACP documentation across the baseline pre-COVID-19 period (September 15, 2019, to March 14, 2020), the COVID-19 wave 1 period (March 15, 2020, to September 14, 2020), and an intervention period (December 15, 2020, to June 14, 2021) at a New York metropolitan area ambulatory network of 22 clinics. All patients 65 years or older who had at least 1 clinic or telehealth visit during any of the 3 study periods were included. Main Outcomes and Measures: The primary outcome was ACP documentation. Results: A total of 14 107 patients (mean [SD] age, 81.0 [8.4] years; 8856 [62.8%] female; and 2248 [15.9%] African American or Hispanic) interacted with clinicians during the pre-COVID-19 period; 12 806 (mean [SD] age, 81.2 [8.5] years; 8047 [62.8%] female; and 1992 [15.6%] African American or Hispanic), during wave 1; and 15 106 (mean [SD] 80.9 [8.3] years; 9543 [63.2%] female; and 2535 [16.8%] African American or Hispanic), during the intervention period. Clinicians documented ACP in 3587 patients (23.8%) during the intervention period compared with 2525 (17.9%) during the pre-COVID-19 period (rate difference [RD], 5.8%; 95% CI, 0.9%-7.9%; P = .01) and 1598 (12.5%) during wave 1 (RD, 11.3%; 95% CI, 6.3%-12.1%; P < .001). Advance care planning was documented in 447 African American patients (30.0%) during the intervention period compared with 233 (18.1%) during the pre-COVID-19 period (RD, 11.9%; 95% CI, 4.1%-15.9%; P < .001) and 130 (11.0%) during wave 1 (RD, 19.1%; 95% CI, 11.7%-21.2%; P < .001). Advance care planning was documented for 222 Hispanic patients (21.2%) during the intervention period compared with 127 (13.2%) during the pre-COVID-19 period (RD, 8.0%; 95% CI, 2.1%-10.9%; P = .004) and 82 (10.2%) during wave 1 (RD, 11.1%; 95% CI, 5.5%-14.5%; P < .001). Conclusions and Relevance: This intervention, implemented during the evolving COVID-19 pandemic, was associated with higher rates of ACP documentation, especially for African American and Hispanic patients. Trial Registration: ClinicalTrials.gov Identifier: NCT04660422.

Likelihood-based deconvolution of bulk gene expression data using single-cell references.

Direct comparison of bulk gene expression profiles is complicated by distinct cell type mixtures in each sample that obscure whether observed differences are actually caused by changes in the expression levels themselves or are simply a result of differing cell type compositions. Single-cell technology has made it possible to measure gene expression in individual cells, achieving higher resolution at the expense of increased noise. If carefully incorporated, such single-cell data can be used to deconvolve bulk samples to yield accurate estimates of the true cell type proportions, thus enabling one to disentangle the effects of differential expression and cell type mixtures. Here, we propose a generative model and a likelihood-based inference method that uses asymptotic statistical theory and a novel optimization procedure to perform deconvolution of bulk RNA-seq data to produce accurate cell type proportion estimates. We show the effectiveness of our method, called RNA-Sieve, across a diverse array of scenarios involving real data and discuss extensions made uniquely possible by our probabilistic framework, including a demonstration of well-calibrated confidence intervals.

Numerous Post-translational Modifications of RNA Polymerase II Subunit Rpb4/7 Link Transcription to Post-transcriptional Mechanisms.

Rpb4/7 binds RNA polymerase II (RNA Pol II) transcripts co-transcriptionally and accompanies them throughout their lives. By virtue of its capacity to interact with key regulators (e.g., RNA Pol II, eIF3, and Pat1) temporally and spatially, Rpb4/7 regulates the major stages of the mRNA life cycle. Here we show that Rpb4/7 can undergo more than 100 combinations of post-translational modifications (PTMs). Remarkably, the Rpb4/7 PTM repertoire changes as the mRNA/Rpb4/7 complex progresses from one stage to the next. These temporal PTMs regulate Rpb4 interactions with key regulators of gene expression that control transcriptional and post-transcriptional stages. Moreover, one mutant type specifically affects mRNA synthesis, whereas the other affects mRNA synthesis and decay; both types disrupt the balance between mRNA synthesis and decay ("mRNA buffering") and the cell's capacity to respond to the environment. We propose that temporal Rpb4/7 PTMs mediate the cross-talk among the various stages of the mRNA life cycle.

The Implementation of Communication Didactics for OB/GYN Residents on the Disclosure of Adverse Perioperative Events.

OBJECTIVE: Communication skills are key components of the patient-physician relationship, yet are not routinely taught during residency. Institutional data demonstrates 75% of residents regularly encounter difficult communication scenarios. This study's objective is to develop and pilot a communications didactic/skills training program for Obstetrics & Gynecology (OB/GYN) residents focused on the disclosure of adverse perioperative events. DESIGN: This was an observational, prospective cohort pilot study. OB/GYN residents completed a 4-hour interactive curriculum using VitalTalk methodology, certified facilitators, and simulated patients in 2019. Participants completed self-assessments of their skill levels at 3 time points: prior to training, immediately post-training, 3-month post-training. Wilcoxon signed rank tests were used to evaluate change in skill levels. SETTING: University-based program in North Carolina. PARTICIPANTS: Participants included all OB/GYN residents from postgraduate years 1-4. Out of 31 residents, 27 participated in the training, 24 completed the immediately post-training survey, and 23 completed the 3-month post-training survey. RESULTS: At baseline, most residents rated their global skill level in communication as novice (37.0%) or advanced beginner (33.3%). Immediately following the intervention, 41.7% of residents ranked their global skill as "competent" and 20.8% as "proficient." These changes were statistically significant (p < 0.001). Notable improvements were seen across multiple variables, including the handling of emotional reactions (p = 0.046). No significant changes were noted between the immediately post-training and 3-month time points, suggesting skill retention. Majority of trainees (78.3%) felt that refresher courses would be useful for skill maintenance. CONCLUSION: A simulation-based formalized communication curriculum is effective for improving OB/GYN resident competence and skill levels in the disclosure of adverse perioperative events. Specific to adverse surgical complications, this curriculum appears feasible for implementation by other training programs. Further work is needed to identify the most beneficial timing and modality of these workshops.

Naturally Occurring tRNAs With Non-canonical Structures.

Transfer RNA (tRNA) is the central molecule in genetically encoded protein synthesis. Most tRNA species were found to be very similar in structure: the well-known cloverleaf secondary structure and L-shaped tertiary structure. Furthermore, the length of the acceptor arm, T-arm, and anticodon arm were found to be closely conserved. Later research discovered naturally occurring, active tRNAs that did not fit the established 'canonical' tRNA structure. This review discusses the non-canonical structures of some well-characterized natural tRNA species and describes how these structures relate to their role in translation. Additionally, we highlight some newly discovered tRNAs in which the structure-function relationship is not yet fully understood.

Leaning In to Serious Illness Care Through Stories and Science.

This volume was planned prior to the COVID-19 pandemic as the North Carolina Institute of Medicine (NCIOM) completed a yearlong task force on serious illness. Beyond the task force report, we wanted to dedicate a special edition of the NCMJ to serious illness issues. We commissioned authors who could discuss the challenges, the current practices, and the extensive personal and professional skills needed to navigate these complicated medical diagnoses that often end in death. Little did we know how timely this would be in light of the current pandemic, and we can only speculate on how the world will look as this is published. Our pre-COVID planning reflected personal experiences we all face with the common denominator of serious illness impacting and shaping our lives. As guest editors, we considered how this NCMJ edition would address personal concerns for you, our reader, as well as ourselves. A physician, a social worker, and a nurse, we each have our stories and we want to invite you to lean in and bring both your head and your heart to this reading. We start by relating two very personal experiences that shaped not only life following loss, but also career choices, clinical practices, and scholarship. As you focus on this journal's content, we hope you will also reflect on the people you care for, as well as the issues we all inevitably face.

The yeast exoribonuclease Xrn1 and associated factors modulate RNA polymerase II processivity in 5' and 3' gene regions.

mRNA levels are determined by the balance between mRNA synthesis and decay. Protein factors that mediate both processes, including the 5'-3' exonuclease Xrn1, are responsible for a cross-talk between the two processes that buffers steady-state mRNA levels. However, the roles of these proteins in transcription remain elusive and controversial. Applying native elongating transcript sequencing (NET-seq) to yeast cells, we show that Xrn1 functions mainly as a transcriptional activator and that its disruption manifests as a reduction of RNA polymerase II (Pol II) occupancy downstream of transcription start sites. By combining our sequencing data and mathematical modeling of transcription, we found that Xrn1 modulates transcription initiation and elongation of its target genes. Furthermore, Pol II occupancy markedly increased near cleavage and polyadenylation sites in xrn1Δ cells, whereas its activity decreased, a characteristic feature of backtracked Pol II. We also provide indirect evidence that Xrn1 is involved in transcription termination downstream of polyadenylation sites. We noted that two additional decay factors, Dhh1 and Lsm1, seem to function similarly to Xrn1 in transcription, perhaps as a complex, and that the decay factors Ccr4 and Rpb4 also perturb transcription in other ways. Interestingly, the decay factors could differentiate between SAGA- and TFIID-dominated promoters. These two classes of genes responded differently to XRN1 deletion in mRNA synthesis and were differentially regulated by mRNA decay pathways, raising the possibility that one distinction between these two gene classes lies in the mechanisms that balance mRNA synthesis with mRNA decay.

Communication Matters: a Survey Study of Communication Didactics in Obstetrics/Gynecology Residency.

BACKGROUND: Delivery of excellent patient care hinges on effective communication. Improved communication between physicians, patients, and colleagues can facilitate shared decision-making and foster successful interprofessional teams. Despite the importance of this skill, little is understood about the status or acceptability of dedicated communication training during obstetrics and gynecology (OB/GYN) residency. OBJECTIVE: To explore the national landscape of dedicated communication didactics during OB/GYN training. METHODS: Residents and program directors (PDs) at ACGME-accredited programs were emailed anonymized surveys. Survey responses pertaining to communication didactics and trainee experiences were evaluated using descriptive statistics and chi-squared tests. RESULTS: Of 143 PDs, 45 responded (31.5%). Although the total number of residents receiving our survey is unattainable, our 215 resident respondents can be estimated to represent at least 4.4% of trainees. 98.1% of residents reported challenging clinical communication at least monthly, with many reporting this weekly (47.9%) and daily (30.0%). A majority of PDs (77.8%) and residents (67.0%) endorsed interest in communication training. 62.2% of programs reported formally teaching communication skills. Certain topics were infrequently taught yet cited by residents as particularly challenging-such as "diffusing conflict" and "angry patient or family members." PDs tended to significantly overestimate trainee competence in conducting difficult conversations with both patients (p = 0.0003) and interdisciplinary colleagues (p < 0.0001), as compared with resident self-assessments. CONCLUSIONS: Residents encounter frequent challenging communications interactions, and often feel inadequately equipped to navigate them. Dedicated didactics may provide a critical component to optimally educating of the next generation of trainees within OB/GYN and more broadly.

SDS-PAGE and Dot Blot Autoradiography: Tools for Quantifying Histidine Kinase Autophosphorylation.

Histidine kinases play a vital role in bacterial signal transduction. However, methods for studying the activity of histidine kinases in vitro are limited in comparison to those for investigating serine, threonine, and tyrosine kinases, largely due to the lability of the phosphoramidate (P-N) bond. Here, we describe two useful methods for quantifying histidine kinase autophosphorylation: SDS-PAGE autoradiography and dot blot autoradiography/scintillation counting.

NosP Modulates Cyclic-di-GMP Signaling in Legionella pneumophila.

Biofilms form when bacteria adhere to a surface and secrete an extracellular polymeric substance. Bacteria embedded within a biofilm benefit from increased resistance to antibiotics, host immune responses, and harsh environmental factors. Nitric oxide (NO) is a signaling molecule that can modulate communal behavior, including biofilm formation, in many bacteria. In many cases, NO-induced biofilm dispersal is accomplished through signal transduction pathways that ultimately lead to a decrease in intracellular cyclic-di-GMP levels. H-NOX (heme nitric oxide/oxygen binding domain) proteins are the best characterized bacterial NO sensors and have been implicated in NO-mediated cyclic-di-GMP signaling, but we have recently discovered a second family of NO-sensitive proteins in bacteria named NosP (NO sensing protein); to date, a clear link between NosP signaling and cyclic-di-GMP metabolism has not been established. Here we present evidence that NosP (Lpg0279) binds to NO and directly affects cyclic-di-GMP production from two-component signaling proteins Lpg0278 and Lpg0277 encoded within the NosP operon. Lpg0278 and Lpg0277 are a histidine kinase and cyclic-di-GMP synthase/phosphodiesterase, respectively, that have already been established as being important in regulating Legionella pneumophila cyclic-di-GMP levels; NosP is thus implicated in regulating cyclic-di-GMP in L. pneumophila.

Nitric Oxide Enters Quorum Sensing via the H-NOX Signaling Pathway in Vibrio parahaemolyticus.

Nitric oxide (NO) plays a major role in the regulation of mammalian biological functions. In recent years, NO has also been implicated in bacterial life cycles, including in the regulation of biofilm formation, and the metabolism of the bacterial second messenger signaling molecule cyclic-di-GMP. In a previous study, we reported the discovery of an NO-responsive quorum sensing (QS) circuit in Vibrio harveyi. Here, we characterize the homologous QS pathway in Vibrio parahaemolyticus. Spectroscopic analysis shows V. parahaemolyticus H-NOX is an NO sensory protein that binds NO in 5/6-coordinated mixed manner. Further, we demonstrate that through ligation to H-NOX, NO inhibits the autophosphorylation activity of an H-NOX-associated histidine kinase (HqsK; H-NOX-associated quorum sensing kinase) that transfers phosphate to the Hpt (histidine-containing phosphotransfer protein) protein LuxU. Indeed, among the three Hpt proteins encoded by V. parahaemolyticus, HqsK transfers phosphate only to the QS-associated phosphotransfer protein LuxU. Finally, we show that NO promotes transcription of the master quorum sensing regulatory gene opaR at low cell density.