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Constitutive JAK-STAT signaling drives the proliferation of most myeloproliferative neoplasms (MPN) and a subset of acute myeloid leukemia (AML), but persistence emerges with chronic exposure to JAK inhibitors. MPN and post-MPN AML are dependent on tyrosine phosphorylation of STATs, but the role of serine STAT1 phosphorylation remains unclear. We previously demonstrated that Mediator kinase inhibitor cortistatin A (CA) reduced proliferation of JAK2-mutant AML in vitro and in vivo and also suppressed CDK8-dependent phosphorylation of STAT1 at serine 727. Here we report that phosphorylation of STAT1 S727 promotes the proliferation of AML cells with JAK-STAT pathway activation. Inhibition of serine phosphorylation by CA promotes growth arrest and differentiation, inhibits colony formation in MPN patient samples and reduces allele burden in MPN mouse models. These results reveal that STAT1 pS727 regulates growth and differentiation in JAK-STAT activated neoplasms and suggest that Mediator kinase inhibition represents a therapeutic strategy to regulate JAK-STAT signaling.

Cortistatin A (CA) is a highly selective inhibitor of the Mediator kinases CDK8 and CDK19. Using CA, we now report a large-scale identification of Mediator kinase substrates in human cells (HCT116). We identified over 16,000 quantified phosphosites including 78 high-confidence Mediator kinase targets within 64 proteins, including DNA-binding transcription factors and proteins associated with chromatin, DNA repair, and RNA polymerase II. Although RNA-seq data correlated with Mediator kinase targets, the effects of CA on gene expression were limited and distinct from CDK8 or CDK19 knockdown. Quantitative proteome analyses, tracking around 7,000 proteins across six time points (0–24 hr), revealed that CA selectively affected pathways implicated in inflammation, growth, and metabolic regulation. Contrary to expectations, increased turnover of Mediator kinase targets was not generally observed. Collectively, these data support Mediator kinases as regulators of chromatin and RNA polymerase II activity and suggest their roles extend beyond transcription to metabolism and DNA repair.

Constitutive JAK-STAT signaling drives the proliferation of most myeloproliferative neoplasms (MPN) and a subset of acute myeloid leukemia (AML), but persistence emerges with chronic exposure to JAK inhibitors. MPN and post-MPN AML are dependent on tyrosine phosphorylation of STATs, but the role of serine STAT1 phosphorylation remains unclear. We previously demonstrated that Mediator kinase inhibitor cortistatin A (CA) reduced proliferation of JAK2-mutant AML in vitro and in vivo and also suppressed CDK8-dependent phosphorylation of STAT1 at serine 727. Here we report that phosphorylation of STAT1 S727 promotes the proliferation of AML cells with JAK-STAT pathway activation. Inhibition of serine phosphorylation by CA promotes growth arrest and differentiation, inhibits colony formation in MPN patient samples and reduces allele burden in MPN mouse models. These results reveal that STAT1 pS727 regulates growth and differentiation in JAK-STAT activated neoplasms and suggest that Mediator kinase inhibition represents a therapeutic strategy to regulate JAK-STAT signaling.

Abstract: The large GTPase dynamin is essential for clathrin‐dependent coated‐vesicle formation. Dynasore is a cell‐permeable small molecule that inhibits the GTPase activity of dynamin1, dynamin2 and Drp1, the mitochondrial dynamin. Dynasore was discovered in a screen of ∼16,000 compounds for inhibitors of the dynamin2 GTPase. Dynasore is a noncompetitive inhibitor of dynamin GTPase activity and blocks dynamin‐dependent endocytosis in cells, including neurons. It is fast acting (seconds) and its inhibitory effect in cells can be reversed by washout. Here we present a detailed synthesis protocol for dynasore, and describe a series of experiments used to analyze the inhibitory effects of dynasore on dynamin in vitro and to study the effects of dynasore on endocytosis in cells. [Copyright &y& Elsevier]

Self-esteem is essential for school-aged children's optimum health. High self-esteem is linked to increased school performance, improved health, and productive behavior. This study reports on the effects of a four-lesson self-esteem enhancement program for six groups of 5th and 6th grade children (N=98). The interactive lessons dealt with an overview of self-esteem, media influences, hiding emotions, and changes in self-esteem. Using a pre-test/ post-test design, Coopersmith's Self-Esteem Inventory (SEI) was used to measure self-esteem. The self-esteem subscales dealing with general and social areas were found to significantly increase over time (p<.05). Girls had more significant changes than boys in the general subscale score and the total self-esteem score. Mean scores showed that children who had friends had more significant changes than those who did not have friends. Children with lower socioeconomic status had lower scores at both the pre and post testing with significance in the general and social subscales. No significance was found related to racial group, family make-up, or the number of household chores or activities. This study supports the effectiveness of a self-esteem enhancement program for girls, those children with friends, and those in lower socioeconomic status. Future research is needed to understand what contributes to the self-esteem of children who report that they do not have friends. [ABSTRACT FROM AUTHOR]

Super-enhancers (SEs), which are composed of large clusters of enhancers densely loaded with the Mediator complex, transcription factors (TFs), and chromatin regulators, drive high expression of genes implicated in cell identity and disease, such as lineage-controlling TFs and oncogenes 1, 2. BRD4 and CDK7 are positive regulators of SE-mediated transcription3,4,5. In contrast, negative regulators of SE-associated genes have not been well described. Here we report that Mediator-associated kinases cyclin-dependent kinase 8 (CDK8) and CDK19 restrain increased activation of key SE-associated genes in acute myeloid leukaemia (AML) cells. We determined that the natural product cortistatin A (CA) selectively inhibited Mediator kinases, had antileukaemic activity in vitro and in vivo, and disproportionately induced upregulation of SE-associated genes in CA-sensitive AML cell lines but not in CA-insensitive cell lines. In AML cells, CA upregulated SE-associated genes with tumour suppressor and lineage-controlling functions, including the TFs CEBPA, IRF8, IRF1 and ETV6 6, 7, 8. The BRD4 inhibitor I-BET151 downregulated these SE-associated genes, yet also has antileukaemic activity. Individually increasing or decreasing expression of these TFs suppressed AML cell growth, providing evidence that leukaemia cells are sensitive to dosage of SE-associated genes. Our results demonstrate that Mediator kinases can negatively regulate SE-associated gene expression in specific cell types and can be pharmacologically targeted as a therapeutic approach to AML.

A prospective, longitudinal study was conducted to examine the natural evolution of levels of fatigue, as measured by the Visual Analogue Scale-F, among a group of 36 primiparous women during the first 6 weeks postpartum. The results revealed that this group of women experienced higher levels of morning fatigue across the 6 weeks than had previously been reported. Their morning fatigue peaked at 4 weeks and then slowly decreased. At the 6th week, the group mean for morning fatigue was 1.42 points (on a 100-point scale) lower than at the 1st week postpartum, suggesting women do not completely recover from the effects of pregnancy, childbirth, and transition to parenthood by 6 weeks postpartum. Maternal age and length of labor were found to be significantly related to the levels of fatigue and energy at various times during the 6 week postpartum period. [ABSTRACT FROM AUTHOR]

Cyclic AMP- (cAMP) and calcium-dependent agonists stimulate chloride secretion through the coordinated activation of distinct apical and basolateral membrane channels and ion transporters in mucosal epithelial cells. Defects in the regulation of Cl– transport across mucosal surfaces occur with cystic fibrosis and V. cholerae infection and can be life threatening. Here we report that secramine B, a small molecule that inhibits activation of the Rho GTPase Cdc42, reduced cAMP-stimulated chloride secretion in the human intestinal cell line T84. Secramine B interfered with a cAMP-gated and Ba2+-sensitive K+ channel, presumably KCNQ1/KCNE3. This channel is required to maintain the membrane potential that sustains chloride secretion. In contrast, secramine B did not affect the Ca2+-mediated chloride secretion pathway, which requires a separate K+ channel activity from that of cAMP. Pirl1, another small molecule structurally unrelated to secramine B that also inhibits Cdc42 activation in vitro, similarly inhibited cAMP-dependent but not Ca2+-dependent chloride secretion. These results suggest that Rho GTPases may be involved in the regulation of the chloride secretory response and identify secramine B an inhibitor of cAMP-dependent K+ conductance in intestinal epithelial cells., Chemistry and Chemical Biology

Background: Asthma is a major chronic disease affecting 8.6% of children in the United States., Objective: The purpose of this research was to assess the use of clinical simulation scenarios using augmented reality technology to evaluate learning outcomes for nurse practitioner students studying pediatric asthma management., Methods: A mixed-methods pilot study was conducted with 2 cohorts of graduate pediatric nurse practitioner students (N=21), with each cohort participating for 2 semesters., Results: Significant improvements in pediatric asthma test scores (P<.001) of student learning were found in both cohorts at posttest in both semesters. Student satisfaction with the augmented reality technology was found to be high. The focus group discussions revealed that the simulation was realistic and helpful for a flipped classroom approach., Conclusions: The study results suggest augmented reality simulation to be valuable in teaching pediatric asthma management content in graduate nursing education., (©Suhasini Kotcherlakota, Peggy Pelish, Katherine Hoffman, Kevin Kupzyk, Patrick Rejda. Originally published in JMIR Nursing Informatics (https://nursing.jmir.org), 02.12.2020.)

This scoping review examined research on transitions among emerging adults, 18- to 30-year-olds, to identify designs, populations, frameworks, transition types, and transition outcomes. A librarian conducted the search, yielding 2067 articles. Using predefined criteria, teams screened abstracts and reviewed articles, with 82% to 100% interrater agreement. Data from the final 160 articles were placed in evidence tables and summarized. Most frequently, the studies had exploratory-descriptive designs (69%), nondiagnosed samples (58%), no theoretical frameworks (58%), developmental transitions (34%), and health-related behavior outcomes (34%). This transition research is in an early stage of knowledge development and would benefit from further theory development.

Clear descriptors of faculty practice and scholarly activities are essential to precisely demonstrate that the faculty practice role meets the standards of academic advancement and to influence academic policy. A description of scholarly clinical activities (1) justifies the benefits of faculty practice by means other than fiscal, (2) provides data for research regarding faculty practice, and (3) provides data to support the nursing profession's political, social, and health care agendas. Guidelines for clinical scholarship are described in this article. A review of relevant literature demonstrates that these guidelines go beyond current models by describing 24 scholarly activities organized into 4 areas: quality, governance, leadership, and knowledge development. Three years of data describing the scholarly activities of a college of nursing faculty engaged in practice are presented to demonstrate the effectiveness of the indicators in achieving these goals. These data can provide valuable information for trend analysis and, through heightened awareness of opportunities, increase faculty clinical scholarship activities.

The purpose of this study was to test the effectiveness of the Tiredness Management Guide (TMG) as a self-care intervention for the management of postpartum fatigue from the second to the sixth week postpartum. An experimental repeated measures analysis of variance design was used with a sample of 68 primiparous mothers. Interaction effects between group membership and time in the hypothesized direction were found for fatigue. When the data were tested from the second through the fourth week, a significant interaction effect was found between group membership and time with the experimental group having lower morning fatigue. Results suggest that using the TMG may reduce levels of morning postpartum fatigue from the second through the fourth week postpartum., (Copyright 2003, Elsevier Science (USA). All rights reserved.)

Having recognized the need to update and modify their orientation program, a PICU nursing management team decided to develop a competency-based program. The goal was to facilitate transition of an orientee into a competent entry-level practitioner. This article outlines the procedure used to develop the initial phases: lists of competencies, competency evaluation worksheet and performance checklists. Later, various learning options and additional evaluation methods will be explored.

The effects of the first child on parent's marital happiness were studied using a sample of 185 subjects. Marital happiness scores were found to be lower in the subjects who were pregnant, those with 5-month-old children, and those with 24-month old children than the childless couples. This study suggests trends indicating that first children may influence marital happiness in a negative manner. These findings will assist nurses in the preparation of families for childbearing and the potential changes that may result.