Integrated lncRNA function upon genomic and epigenomic regulation.

Although some long noncoding (lnc)RNAs are known since the 1950s, the past 25 years have uncovered myriad lncRNAs with diverse sequences, structures, and functions. The advent of high-throughput and sensitive technologies has further uncovered the vast heterogeneity of lncRNA-interacting molecules and patterns of expressed lncRNAs. We propose a unifying functional theme for the expansive family of lncRNAs. At an elementary level, the genomic program of gene expression is elicited via canonical transcription and post-transcriptional mRNA assembly, turnover, and translation. Building upon this regulation, an epigenomic program refines the basic genomic control by modifying chromatin architecture as well as DNA and RNA chemistry. Superimposed over the genomic and epigenomic programs, lncRNAs create an additional regulatory dimension: by interacting with the proteins and nucleic acids that regulate gene expression in the nucleus and cytoplasm, lncRNAs help establish robust, nimble, and specific transcriptional and post-transcriptional control. We describe our present understanding of lncRNA-coordinated control of protein programs and cell fate and discuss challenges and opportunities as we embark on the next 25 years of lncRNA discovery.

Identification and functional analysis of senescent cells in the cardiovascular system using omics approaches.

Cardiovascular disease (CVD) is a leading cause of morbidity and mortality worldwide, and senescent cells have emerged as key contributors to its pathogenesis. Senescent cells exhibit cell cycle arrest and secrete a range of proinflammatory factors, termed the senescence-associated secretory phenotype (SASP), which promotes tissue dysfunction and exacerbates CVD progression. Omics technologies, specifically transcriptomics and proteomics, offer powerful tools to uncover and define the molecular signatures of senescent cells in cardiovascular tissue. By analyzing the comprehensive molecular profiles of senescent cells, omics approaches can identify specific genetic alterations, gene expression patterns, protein abundances, and metabolite levels associated with senescence in CVD. These omics-based discoveries provide insights into the mechanisms underlying senescence-induced cardiovascular damage, facilitating the development of novel diagnostic biomarkers and therapeutic targets. Furthermore, integration of multiple omics data sets enables a systems-level understanding of senescence in CVD, paving the way for precision medicine approaches to prevent or treat cardiovascular aging and its associated complications.

Reduction of lamin B receptor levels by miR-340-5p disrupts chromatin, promotes cell senescence and enhances senolysis.

A major stress response influenced by microRNAs (miRNAs) is senescence, a state of indefinite growth arrest triggered by sublethal cell damage. Here, through bioinformatic analysis and experimental validation, we identified miR-340-5p as a novel miRNA that foments cellular senescence. miR-340-5p was highly abundant in diverse senescence models, and miR-340-5p overexpression in proliferating cells rendered them senescent. Among the target mRNAs, miR-340-5p prominently reduced the levels of LBR mRNA, encoding lamin B receptor (LBR). Loss of LBR by ectopic overexpression of miR-340-5p derepressed heterochromatin in lamina-associated domains, promoting the expression of DNA repetitive elements characteristic of senescence. Importantly, overexpressing miR-340-5p enhanced cellular sensitivity to senolytic compounds, while antagonization of miR-340-5p reduced senescent cell markers and engendered resistance to senolytic-induced cell death. We propose that miR-340-5p can be exploited for removing senescent cells to restore tissue homeostasis and mitigate damage by senescent cells in pathologies of human aging.

AUF1 ligand circPCNX reduces cell proliferation by competing with p21 mRNA to increase p21 production.

Mammalian circRNAs can influence different cellular processes by interacting with proteins and other nucleic acids. Here, we used ribonucleoprotein immunoprecipitation (RIP) analysis to identify systematically the circRNAs associated with the cancer-related protein AUF1. Among the circRNAs interacting with AUF1 in HeLa (human cervical carcinoma) cells, we focused on hsa_circ_0032434 (circPCNX), an abundant target of AUF1. Overexpression of circPCNX specifically interfered with the binding of AUF1 to p21 (CDKN1A) mRNA, thereby promoting p21 mRNA stability and elevating the production of p21, a major inhibitor of cell proliferation. Conversely, silencing circPCNX increased AUF1 binding to p21 mRNA, reducing p21 production and promoting cell division. Importantly, eliminating the AUF1-binding region of circPCNX abrogated the rise in p21 levels and rescued proliferation. Therefore, we propose that the interaction of circPCNX with AUF1 selectively prevents AUF1 binding to p21 mRNA, leading to enhanced p21 mRNA stability and p21 protein production, thereby suppressing cell growth.

The Association Between Work as a Calling and Turnover Among Early Childhood Education Professionals.

Turnover of early childhood education (ECE) professionals negatively impacts program costs, staff morale, and relationships with children. We determined whether the presence of work as a calling was associated with less intention to leave the ECE field. From an online survey administered to 265 ECE professionals in Pennsylvania, a calling score based on the Calling and Vocation Questionnaire was used to create sample-defined tertiles of low (< 38), medium (38-44), and high (> 44) presence of calling. Those intending to leave the ECE field reported that, given the option, they would most likely "find a position or get training in a completely different field," or "stop work, stay home, or retire." Analysis was restricted to 194 respondents currently employed in ECE and under age 60, of whom 94.8% were female and 53.9% were non-Hispanic White. After adjusting for race/ethnicity and workplace stress, the prevalence (95% CI) of intention to leave decreased as calling increased, from low (28.6% [17.8%, 38.4%]) to medium (12.2% [4.3%, 20.1%]) to high (9.1% [1.5%, 16.6%]). The presence of call was associated with less intention to leave the ECE field. Identifying, building, and sustaining call among ECE professionals may decrease turnover.

The Association Between Early Childhood Education Professionals' Perception of Workplace Spirituality and Their Intention to Stay in Their Current Program.

Turnover in the US early childhood education (ECE) workforce is associated with worse outcomes for children. Greater workplace spirituality, or the perception of meaningful work, sense of community, and alignment with organizational values, is associated with reduced turnover. However, this association has not been examined in ECE professionals. We administered an online survey to 265 ECE professionals in Pennsylvania (US) in the spring of 2021. Respondents were asked about their intention to stay in their current program, if given the option to leave. Workplace spirituality was measured with a 21-item scale assessing the dimensions of meaningful work, sense of community, and alignment with organizational values. The survey was completed by 246 (92.8%), and data were analyzed for 232 respondents. Of these, 94.8% were female, 54.4% were non-Hispanic White, and 70.7% had a bachelor's or graduate degree. The prevalence of intention to stay was 33.2%. After adjusting for all covariates, including gender, age, race/ethnicity, education, job position, workplace stress, and economic hardships, the prevalence (95% confidence interval [CI]) of intention to stay increased across tertiles of workplace spirituality from low to medium to high: 16.4% (7.9%, 24.9%) to 38.6% (28.4%, 48.8%) to 43.7% (32.1%, 55.3%), respectively. ECE professionals who perceived greater workplace spirituality were more likely to report they intended to stay in their current program. Turnover in the ECE workforce could potentially be reduced through efforts to increase a sense of meaning and community at work and to align the values of ECE programs with those who work in them. Supplementary Information: The online version contains supplementary material available at 10.1007/s10643-023-01506-7.

Factors and Pathways Modulating Endothelial Cell Senescence in Vascular Aging.

Aging causes a progressive decline in the structure and function of organs. With advancing age, an accumulation of senescent endothelial cells (ECs) contributes to the risk of developing vascular dysfunction and cardiovascular diseases, including hypertension, diabetes, atherosclerosis, and neurodegeneration. Senescent ECs undergo phenotypic changes that alter the pattern of expressed proteins, as well as their morphologies and functions, and have been linked to vascular impairments, such as aortic stiffness, enhanced inflammation, and dysregulated vascular tone. Numerous molecules and pathways, including sirtuins, Klotho, RAAS, IGFBP, NRF2, and mTOR, have been implicated in promoting EC senescence. This review summarizes the molecular players and signaling pathways driving EC senescence and identifies targets with possible therapeutic value in age-related vascular diseases.

Diabetes distress and glycaemic control in young adults with type 1 diabetes: Associations by use of insulin pumps and continuous glucose monitors.

AIMS: We determined whether high diabetes distress (DD) in young adults with type 1 diabetes was associated with higher glycated haemoglobin (HbA1c ) levels and whether this association was similar among those who were and were not using diabetes devices (insulin pumps and/or continuous glucose monitors [CGMs]). METHODS: In 2017, an online survey was completed by 423 of 743 (57%) young adults (19-31 years) with type 1 diabetes receiving care at a specialty clinic in New York City. HbA1c level was the primary outcome measure, and high DD (Diabetes Distress Scale score ≥3) was the primary exposure. Associations were adjusted for sociodemographic covariates. RESULTS: Of the 419 respondents with complete DD data, 59% were female and 69% were non-Hispanic white. Both devices (pump and CGM) were used by 35%, either device by 42% and neither device by 24%. The mean (SD) HbA1c was 64 (19) mmol/mol (8.0 [1.7] %) and 24% had high DD. The adjusted mean (95% confidence interval) HbA1c was 10 (6, 14) mmol/mol (0.9 [0.5, 1.2] %) greater in those with high DD than in those without it. This HbA1c difference associated with high DD was similar regardless of device use: 9 (3, 15) mmol/mol (0.8 [0.3, 1.4] %) greater among those using both devices; and 9 (-0.9, 18) mmol/mol (0.8 [-0.1, 1.7] %) greater among those using neither device. CONCLUSIONS: High DD in young adults requires more attention because it is associated with higher HbA1c levels, even among those using insulin pumps and CGMs.

The interaction of adverse childhood experiences and gender as risk factors for depression and anxiety disorders in US adults: a cross-sectional study.

BACKGROUND: Exposure to adverse childhood experiences (ACEs) and being female are distinct risk factors for having a major depressive episode (MDE) or an anxiety disorder (AD) in adulthood, but it is unclear whether these two risk factors are synergistic. The purpose of this study was to determine whether exposure to ACEs and being female are more than additive (synergistic) in their association with MDE and AD in US adults. METHODS: We pooled cross-sectional survey data in the Midlife in the United States study from two nationally-representative cohorts of English-speaking US adults. Data from the first cohort were collected in 2004-2006 and from the second in 2011-2014. Data from both cohorts included the 12-month prevalence of MDE and AD (generalized anxiety disorder or panic disorder) assessed with the Composite International Diagnostic Interview Short Form, gender (here termed female and male), and the count of five categories of exposure to ACEs: physical, sexual, or emotional abuse; household alcohol or substance abuse; and parental separation or divorce. RESULTS: Of the 5834 survey respondents, 4344 (74.5%) with complete data on ACEs were included in the analysis. Mean (SD) age was 54.1 (13.8) years and 53.9% were female. The prevalences of MDE, AD, and exposure to 3-5 categories of ACEs were 13.7, 10.0, and 12.5%, respectively. After adjusting for covariates (age, race, and current and childhood socioeconomic disadvantage), for those with both risk factors (female and 3-5 ACEs) the prevalence of MDE was 26.9%. This was 10.2% (95% CI: 1.8, 18.5%) higher than the expected prevalence based on the additive associations of the two risk factors. The adjusted prevalence of AD among females with 3-5 ACEs was 21.9%, which was 11.4% (95% CI: 4.0, 18.9%) higher than the expected prevalence. CONCLUSIONS: For both MDE and AD, there was synergy between the two risk factors of exposure to ACEs and being female. Identification and treatment of MDE and AD may benefit from understanding the mechanisms involved in the synergistic interaction of gender with ACEs.

RNA stability protein ILF3 mediates cytokine-induced angiogenesis.

Interleukin enhancer-binding factor 3 (ILF3), an RNA-binding protein, is best known for its role in innate immunity by participation in cellular antiviral responses. A role for ILF3 in angiogenesis is unreported. ILF3 expression in CD31+ capillaries of hypoxic cardiac tissue was detected by immunohistochemistry. Proangiogenic stimuli induce ILF3 mRNA and protein expression in cultured human coronary artery endothelial cells (hCAECs). Angiogenic indices, including proliferation, migration, and tube formation, are all significantly reduced in hCAECs when ILF3 is knocked down using small interfering RNA (siRNA), but are significantly increased when ILF3 is overexpressed using adenovirus. Protein and mRNA abundance of several angiogenic factors including CXCL1, VEGF, and IL-8 are decreased when ILF3 is knocked down by siRNA. These factors are increased when ILF3 is overexpressed by adenovirus. ILF3 is phosphorylated and translocates from the nucleus to the cytoplasm in response to angiogenic stimuli. Proangiogenic transcripts containing adenine and uridine-rich elements were bound to ILF3 through RNA immunoprecipitation. ILF3 stabilizes proangiogenic transcripts including VEGF, CXCL1, and IL-8 in hCAECs. Together these data suggest that in endothelial cells, the RNA stability protein, ILF3, plays a novel and central role in angiogenesis. Our working hypothesis is that ILF3 promotes angiogenesis through cytokine-inducible mRNA stabilization of proangiogenic transcripts.-Vrakas, C. N., Herman, A. B., Ray, M., Kelemen, S. E., Scalia, R., Autieri, M. V. RNA stability protein ILF3 mediates cytokine-induced angiogenesis.

Regulation of Stress Granule Formation by Inflammation, Vascular Injury, and Atherosclerosis.

OBJECTIVE: Stress granules (SGs) are dynamic cytoplasmic aggregates containing mRNA, RNA-binding proteins, and translation factors that form in response to cellular stress. SGs have been shown to contribute to the pathogenesis of several human diseases, but their role in vascular diseases is unknown. This study shows that SGs accumulate in vascular smooth muscle cells (VSMCs) and macrophages during atherosclerosis. Approach and Results: Immunohistochemical analysis of atherosclerotic plaques from LDLR-/- mice revealed an increase in the stress granule-specific markers Ras-G3BP1 (GTPase-activating protein SH3 domain-binding protein) and PABP (poly-A-binding protein) in intimal macrophages and smooth muscle cells that correlated with disease progression. In vitro, PABP+ and G3BP1+ SGs were rapidly induced in VSMC and bone marrow-derived macrophages in response to atherosclerotic stimuli, including oxidized low-density lipoprotein and mediators of mitochondrial or oxidative stress. We observed an increase in eIF2α (eukaryotic translation initiation factor 2-alpha) phosphorylation, a requisite for stress granule formation, in cells exposed to these stimuli. Interestingly, SG formation, PABP expression, and eIF2α phosphorylation in VSMCs is reversed by treatment with the anti-inflammatory cytokine interleukin-19. Microtubule inhibitors reduced stress granule accumulation in VSMC, suggesting cytoskeletal regulation of stress granule formation. SG formation in VSMCs was also observed in other vascular disease pathologies, including vascular restenosis. Reduction of SG component G3BP1 by siRNA significantly altered expression profiles of inflammatory, apoptotic, and proliferative genes. CONCLUSIONS: These results indicate that SG formation is a common feature of the vascular response to injury and disease, and that modification of inflammation reduces stress granule formation in VSMC.

The association between dispositional mindfulness and glycemic control in type 1 diabetes during early adulthood: Differences by age and adverse childhood experiences.

BACKGROUND: The study objective was to determine whether higher levels of dispositional mindfulness were associated with lower HbA1c levels among young adults with type 1 diabetes (T1D) and whether this association differed by age or exposure to adverse childhood experiences (ACEs). METHODS: An online cross-sectional survey, called T1 Flourish, was completed in 2017 by 423 of 743 (56.9%) young adults (19-31 years) with T1D receiving outpatient care at a diabetes specialty clinic in New York City. HbA1c levels were abstracted from medical records. Respondents were categorized by age, high and low dispositional mindfulness (median split on Cognitive and Affective Mindfulness Scale-Revised), and exposure to any of 10 ACEs. RESULTS: Respondents had a mean (SD) HbA1c of 64 (18) mmol/mol [8.0 (1.7)%]; 59.3% were female and 69.4% were non-Hispanic white. The covariate-adjusted association between dispositional mindfulness and HbA1c differed by age group and ACEs. Among 27- to 31-year-olds, those with high mindfulness had HbA1c levels that were 8 mmol/mol [0.7%] lower (95% confidence interval, 2-13 mmol/mol [0.2-1.2%]) than those with low mindfulness, and this association tended to be stronger in those with ≥1 ACEs. Weaker, non-significant associations in the same direction occurred in 23- to 26-year-olds. Among 19- to 22-year-olds, those with high mindfulness and no ACEs tended to have higher HbA1c levels. CONCLUSIONS: In young adults with T1D, higher mindfulness was significantly associated with lower HbA1c only among 27- to 31-year-olds. In early adulthood, the impact of mindfulness-based interventions on glycemic control may vary by age and childhood trauma history.

Drowsy Driving, Sleep Duration, and Chronotype in Adolescents.

OBJECTIVE: To determine whether self-reported drowsy driving was associated with an evening chronotype, a biologically-based difference in circadian sleep-wake timing, and shorter school-night sleep duration in a sample of high school drivers. STUDY DESIGN: Cross-sectional observational data were obtained from an online survey in spring 2015 of 431 drivers, age 15.5-18.7 years, attending Fairfax County (Virginia) Public schools. Drowsy driving was defined as having ever "driven a car or motor vehicle while feeling drowsy" in the last year. School-night sleep duration was calculated from school-night bedtime and wake time. Those with scores in the lower and upper tertiles of the Morningness-Eveningness Scale for Children were designated as having an evening or morning chronotype, respectively. RESULTS: Among survey respondents, 63.1% drove at least several times a week and 47.6% reported drowsy driving. The covariate-adjusted prevalence of drowsy driving was 13.9% (95% CI 3.0%-24.9%) higher in students who slept <7 hours on school-nights than in those who slept 8 or more hours. Compared with those with a morning chronotype, the adjusted prevalence of drowsy driving was 15.2% (95% CI 4.5%-25.9%) higher among those with an evening chronotype. CONCLUSION: Among adolescent drivers, both an evening chronotype and shorter school-night sleep duration were associated with more frequent reports of drowsy driving. Interventions to improve the timing and duration of nighttime sleep in adolescents may reduce the occurrence of drowsy driving.

Genetic Deletion of IL-19 (Interleukin-19) Exacerbates Atherogenesis in Il19-/-×Ldlr-/- Double Knockout Mice by Dysregulation of mRNA Stability Protein HuR (Human Antigen R).

OBJECTIVE: To test the hypothesis that loss of IL-19 (interleukin-19) exacerbates atherosclerosis. APPROACH AND RESULTS: Il19-/- mice were crossed into Ldlr-/- (low-density lipoprotein receptor knock out) mice. Double knockout (dKO) mice had increased plaque burden in aortic arch and root compared with Ldlr-/- controls after 14 weeks of high-fat diet (HFD). dKO mice injected with 10 ng/g per day rmIL-19 had significantly less plaque compared with controls. qRT-PCR and Western blot analysis revealed dKO mice had increased systemic and intraplaque polarization of T cells and macrophages to proinflammatory Th1 and M1 phenotypes, and also significantly increased TNF (tumor necrosis factor)-α expression in spleen and aortic arch compared with Ldlr-/- controls. Bone marrow transplantation suggests that immune cells participate in IL-19 protection. Bone marrow-derived macrophages and vascular smooth muscle cells isolated from dKO mice had a significantly greater expression of inflammatory cytokine mRNA and protein compared with controls. Spleen and aortic arch from dKO mice had significantly increased expression of the mRNA stability protein HuR (human antigen R). Bone marrow-derived macrophage and vascular smooth muscle cell isolated from dKO mice also had greater HuR abundance. HuR stabilizes proinflammatory transcripts by binding AU-rich elements in the 3' untranslated region. Cytokine and HuR mRNA stability were increased in dKO bone marrow-derived macrophage and vascular smooth muscle cell, which was rescued by addition of IL-19 to these cells. IL-19-induced expression of miR133a, which targets and reduced HuR abundance; miR133a levels were lower in dKO mice compared with controls. CONCLUSIONS: These data indicate that IL-19 is an atheroprotective cytokine which decreases the abundance of HuR, leading to reduced inflammatory mRNA stability.

The association of fatigue with dispositional mindfulness: relationships by levels of depressive symptoms, sleep quality, childhood adversity, and chronic medical conditions.

Although mindfulness-based interventions may be effective in addressing the common symptom of fatigue, no population-based studies have examined the relationship between mindfulness and fatigue. We determined whether higher levels of dispositional mindfulness were associated with lower levels of fatigue. Cross-sectional data were obtained through the Pennsylvania Head Start Staff Wellness Survey, a 2012 web-based survey in which 2199 of 3375 (65%) eligible staff participated. The analytic sample was restricted to the 2083 female respondents with complete data on dispositional mindfulness (Cognitive and Affective Mindfulness Scale-Revised) and fatigue (Fatigue Severity Scale). We determined the mean covariate-adjusted fatigue scores in each quartile of dispositional mindfulness. This relationship was examined in the overall sample and within subgroups defined by levels of four variables: depressive symptoms, poor sleep quality, childhood adversity, and chronic medical conditions. The sample was 86% non-Hispanic White, and 61% had a bachelor's or more advanced degree. The mean (SD) Fatigue Severity Scale score was 3.3 (1.3). The adjusted mean fatigue score decreased significantly and in a graded manner across higher quartiles of mindfulness, with the adjusted fatigue score 1.4 points lower (95% confidence interval: -1.5, -1.2) among those in the highest quartile of dispositional mindfulness compared to the lowest. This significant graded relationship was present within each subgroup examined, and there was not a statistically significant interaction between dispositional mindfulness and any subgroup variable. Future trials of mindfulness-based interventions should consider assessing the outcome of fatigue in both clinical and non-clinical populations.

Induction of MiR133a expression by IL-19 targets LDLRAP1 and reduces oxLDL uptake in VSMC.

The transformation of vascular smooth muscle cells [VSMC] into foam cells leading to increased plaque size and decreased stability is a key, yet understudied step in atherogenesis. We reported that Interleukin-19 (IL-19), a novel, anti-inflammatory cytokine, attenuates atherosclerosis by anti-inflammatory effects on VSMC. In this work we report that IL-19 induces expression of miR133a, a muscle-specific miRNA, in VSMC. Although previously unreported, we report that miR133a can target and reduce mRNA abundance, mRNA stability, and protein expression of Low Density Lipoprotein Receptor Adaptor Protein 1, (LDLRAP1), an adaptor protein which functions to internalize the LDL receptor. Mutations in this gene lead to LDL receptor malfunction and cause the Autosomal Recessive Hypercholesterolemia (ARH) disorder in humans. Herein we show that IL-19 reduces lipid accumulation in VSMC, and LDLRAP1 expression and oxLDL uptake in a miR133a-dependent mechanism. We show that LDLRAP1 is expressed in plaque and neointimal VSMC of mouse and human injured arteries. Transfection of miR133a and LDLRAP1 siRNA into VSMC reduces their proliferation and uptake of oxLDL. miR133a is significantly increased in plasma from hyperlipidemic compared with normolipidemic patients. Expression of miR133a in IL-19 stimulated VSMC represents a previously unrecognized link between vascular lipid metabolism and inflammation, and may represent a therapeutic opportunity to combat vascular inflammatory diseases.

Inflammation-regulated mRNA stability and the progression of vascular inflammatory diseases.

Cardiovascular disease remains a major medical and socioeconomic burden in developed and developing societies, and will increase with an aging and increasingly sedentary society. Vascular disease and atherosclerotic vascular syndromes are essentially inflammatory disorders, and transcriptional and post-transcriptional processes play essential roles in the ability of resident vascular and inflammatory cells to adapt to environmental stimuli. The regulation of mRNA translocation, stability, and translation are key processes of post-transcriptional regulation that permit these cells to rapidly respond to inflammatory stimuli. For the most part, these processes are controlled by elements in the 3'-UTR of labile, proinflammatory transcripts. Since proinflammatory transcripts almost exclusively contain AU-rich elements (AREs), this represents a tightly regulated and specific mechanism for initiation and maintenance of the proinflammatory phenotype. RNA-binding proteins (RBPs) recognize cis elements in 3'-UTR, and regulate each of these processes, but there is little literature exploring the concept that RBPs themselves can be directly regulated by inflammatory stimuli. Conceptually, inflammation-responsive RBPs represent an attractive target of rational therapies to combat vascular inflammatory syndromes. Herein we briefly describe the cellular and molecular etiology of atherosclerosis, and summarize our current understanding of RBPs and their specific roles in regulation of inflammatory mRNA stability. We also detail RBPs as targets of current anti-inflammatory modalities and how this may translate into better treatment for vascular inflammatory diseases.

The Bacterial iprA Gene Is Conserved across Enterobacteriaceae, Is Involved in Oxidative Stress Resistance, and Influences Gene Expression in Salmonella enterica Serovar Typhimurium.

UNLABELLED: The iprA gene (formerly known as yaiV or STM0374) is located in a two-gene operon in the Salmonella enterica serovar Typhimurium genome and is associated with altered expression during spaceflight and rotating-wall-vessel culture conditions that increase virulence. However, iprA is uncharacterized in the literature. In this report, we present the first targeted characterization of this gene, which revealed that iprA is highly conserved across Enterobacteriaceae We found that S Typhimurium, Escherichia coli, and Enterobacter cloacae ΔiprA mutant strains display a multi-log-fold increase in oxidative stress resistance that is complemented using a plasmid-borne wild-type (WT) copy of the S Typhimurium iprA gene. This observation was also associated with increased catalase activity, increased S Typhimurium survival in macrophages, and partial dependence on the katE gene and full dependence on the rpoS gene. Our results indicate that IprA protein activity is sensitive to deletion of the N- and C-terminal 10 amino acids, while a region that includes amino acids 56 to 80 is dispensable for activity. RNA sequencing (RNA-Seq) analysis revealed several genes altered in expression in the S Typhimurium ΔiprA mutant strain compared to the WT, including those involved in fimbria formation, spvABCD-mediated virulence, ethanolamine utilization, the phosphotransferase system (PTS) transport, and flagellin phase switching from FlgB to FliC (likely a stochastic event) and several genes of hypothetical or putative function. IMPORTANCE: Overall, this work reveals that the conserved iprA gene measurably influences bacterial biology and highlights the pool of currently uncharacterized genes that are conserved across bacterial genomes. These genes represent potentially useful targets for bacterial engineering, vaccine design, and other possible applications.

The Association of Childhood Parental Connection With Adult Flourishing and Depressive Symptoms.

OBJECTIVES: To determine whether a common measure of childhood emotional neglect, scored instead as a continuous measure of increasing parental connection, is associated with adult flourishing and depressive symptoms, and to compare the magnitude of these 2 associations. METHODS: We pooled cross-sectional survey data from the Midlife in the United States study, collected from 2 national cohorts (2004-2006 and 2011-2014) of English-speaking, US adults, aged 25 to 74 years. Using the 5-item emotional neglect subscale of the Childhood Trauma Questionnaire, a score of increasing childhood parental connection was created by not reverse-scoring responses. The adult outcomes were standardized scores of flourishing, from Ryff's Psychological Well-Being Scale, and depressive symptoms, from the Center for Epidemiologic Studies Depression Scale. RESULTS: Data were available for 2079 of 2118 participants (98.2%). The mean (SD) age was 53.1 (12.6) years and 54.6% were female. After adjusting for covariates (age, gender, race and ethnicity, marital status, chronic disease, socioeconomic disadvantage), the adult flourishing score was 0.74 (95% confidence interval 0.63-0.86) SD units higher in those in the highest quartile of childhood parental connection compared with the lowest, whereas the depressive symptoms score was lower by a similar magnitude (-0.65 [95% confidence interval -0.77 to -0.54] SD units). CONCLUSIONS: When emotional neglect is reframed as parental connection, it has associations with adult flourishing and depressive symptoms that are of similar magnitude but opposite direction. Clinicians and researchers should consider the more positive and aspirational frame of parental connection and its potential contribution to life course flourishing.

Trauma-Informed Undergraduate Medical Education: A Pathway to Flourishing with Adversity by Enhancing Psychological Safety.

We describe the Life Experiences Curriculum (LEC), which attempts to integrate medical student well-being with trauma-informed medical education. The long-term goal of LEC is to help medical students flourish with adversity and trauma, where flourishing refers to having a sense of purpose that arises from awareness of one's strengths and limitations, shaped by life experiences. The short-term goal of LEC is to develop students' relational capacities, such as acceptance and awareness of self and others, while building and maintaining students' psychological safety. We describe the conceptual rationale for these goals and the curriculum's development, implementation, evaluation, and limitations. The curriculum extends over four years and involves a preclinical seminar and students' individual and group reflection sessions with LEC faculty. The seminar addresses the coexistence of trauma and flourishing across life experiences, as well as how safety in relationships is impaired by traumatic experiences and must be restored for healing and growth. The physician faculty have no role in student evaluation and co-lead all LEC activities. LEC is intended to provide students with new language for understanding the process of trauma and flourishing in both individuals and systems and to build and sustain students' relational capacities. There are ongoing efforts to re-imagine self-care as communal-care in which care and support are given and received in a community of students and faculty. Such a model may help build the relational capacities needed to deliver trauma-informed care and also promote flourishing with adversity in healers and in those seeking to be healed.