Barriers to Reproductive Healthcare for Women With Opioid Use Disorder.

The health impact of opioid use disorder on women and infant health alongside persistent rates of unintended pregnancy calls for better targeted reproductive healthcare for all women, especially those receiving treatment for opioid treatment disorder and decreasing barriers to care. This cross-sectional mixed-methods study explored the reproductive intentions and contraceptive practices of women (N = 50) in medication-assisted treatment of opioid use disorder with a focus on knowledge and use of long-acting reversible contraception (LARC), specifically intrauterine devices and implants. Eighty-four percent of the 50 women interviewed had experienced at least 1 unintended pregnancy, and 30% were using contraceptive methods with high failure rates. Although approximately 75% of women indicated awareness of both forms of LARCs, only 6% reported current or past use of either device. Qualitative results found the greatest barriers to the uptake of LARC were women's expressed fears of complications and inaccurate information from family, friends, and acquaintances. Increasing awareness of the benefits of LARC as ideal contraception for women who may desire a future pregnancy is important for improving reproductive health for women receiving treatment of opioid use disorder.

PAK6 targets to cell-cell adhesions through its N-terminus in a Cdc42-dependent manner to drive epithelial colony escape.

The six serine/threonine kinases in the p21-activated kinase (PAK) family are important regulators of cell adhesion, motility and survival. PAK6, which is overexpressed in prostate cancer, was recently reported to localize to cell-cell adhesions and to drive epithelial cell colony escape. Here we report that PAK6 targeting to cell-cell adhesions occurs through its N-terminus, requiring both its Cdc42/Rac interactive binding (CRIB) domain and an adjacent polybasic region for maximal targeting efficiency. We find PAK6 localization to cell-cell adhesions is Cdc42-dependent, as Cdc42 knockdown inhibits PAK6 targeting to cell-cell adhesions. We further find the ability of PAK6 to drive epithelial cell colony escape requires kinase activity and is disrupted by mutations that perturb PAK6 cell-cell adhesion targeting. Finally, we demonstrate that all type II PAKs (PAK4, PAK5 and PAK6) target to cell-cell adhesions, albeit to differing extents, but PAK1 (a type I PAK) does not. Notably, the ability of a PAK isoform to drive epithelial colony escape correlates with its targeting to cell-cell adhesions. We conclude that PAKs have a broader role in the regulation of cell-cell adhesions than previously appreciated.

Safety of Rolled and Folded Cotton Blankets Warmed in 130°F and 200°F Cabinets.

PURPOSE: In 2014, ECRI recommended that blanket warming cabinets be set at a maximum temperature of 130°F because of safety concerns with warmed rolled and folded blankets. We could find no research to support this recommendation. The purpose of this study was to measure skin temperatures and thermal comfort in healthy volunteers before and after application of folded and rolled dry cotton blankets warmed in 130°F or 200°F cabinets. DESIGN: Randomized, descriptive, and comparative study. METHODS: Participants (n = 20) received two blankets (one rolled and one folded) from warming cabinets set at 130°F or 200°F. Folded blankets were applied to the back and rolled to the neck. Skin temperatures and thermal comfort were obtained at fixed time intervals. FINDINGS: Skin temperatures from blankets in the 200°F cabinet were greater than those in the 130°F cabinet. No skin temperatures reached temperature and/or duration thresholds for dermal injury. CONCLUSIONS: This study provides supportive evidence that warming cabinets may be set at a maximum of 200°F without compromising patient safety.

Signaling, Regulation, and Specificity of the Type II p21-activated Kinases.

The p21-activated kinases (PAKs) are a family of six serine/threonine kinases that act as key effectors of RHO family GTPases in mammalian cells. PAKs are subdivided into two groups: type I PAKs (PAK1, PAK2, and PAK3) and type II PAKs (PAK4, PAK5, and PAK6). Although these groups are involved in common signaling pathways, recent work indicates that the two groups have distinct modes of regulation and have both unique and common substrates. Here, we review recent insights into the molecular level details that govern regulation of type II PAK signaling. We also consider mechanisms by which signal transduction is regulated at the level of substrate specificity. Finally, we discuss the implications of these studies for clinical targeting of these kinases.

Selective inhibition of BET bromodomains.

Epigenetic proteins are intently pursued targets in ligand discovery. So far, successful efforts have been limited to chromatin modifying enzymes, or so-called epigenetic 'writers' and 'erasers'. Potent inhibitors of histone binding modules have not yet been described. Here we report a cell-permeable small molecule (JQ1) that binds competitively to acetyl-lysine recognition motifs, or bromodomains. High potency and specificity towards a subset of human bromodomains is explained by co-crystal structures with bromodomain and extra-terminal (BET) family member BRD4, revealing excellent shape complementarity with the acetyl-lysine binding cavity. Recurrent translocation of BRD4 is observed in a genetically-defined, incurable subtype of human squamous carcinoma. Competitive binding by JQ1 displaces the BRD4 fusion oncoprotein from chromatin, prompting squamous differentiation and specific antiproliferative effects in BRD4-dependent cell lines and patient-derived xenograft models. These data establish proof-of-concept for targeting protein-protein interactions of epigenetic 'readers', and provide a versatile chemical scaffold for the development of chemical probes more broadly throughout the bromodomain family.

Soluble IL7Rα potentiates IL-7 bioactivity and promotes autoimmunity.

Human soluble interleukin-7 receptor (sIL7R)α circulates in high molar excess compared with IL-7, but its biology remains unclear. We demonstrate that sIL7Rα has moderate affinity for IL-7 but does not bind thymic stromal lymphopoietin. Functionally, sIL7Rα competes with cell-associated IL-7 receptor to diminish excessive IL-7 consumption and, thus, enhances the bioactivity of IL-7 when the cytokine is limited, as it is presumed to be in vivo. IL-7 signaling in the presence of sIL7Rα also diminishes expression of CD95 and suppressor of cytokine signaling 1, both regulatory molecules. Murine models confirm diminished consumption of IL-7 in the presence of sIL7Rα and also demonstrate a potentiating effect of sIL7Rα on IL-7-mediated homeostatic expansion and experimental autoimmune encephalomyelitis exacerbation. In multiple sclerosis and several other autoimmune diseases, IL7R genotype influences susceptibility. We measured increased sIL7Rα levels, as well as increased IL-7 levels, in multiple sclerosis patients with the predisposing IL7R genotype, consistent with diminished IL-7 consumption in vivo. This work demonstrates that sIL7Rα potentiates IL-7 bioactivity and provides a basis to explain the increased risk of autoimmunity observed in individuals with genotype-induced elevations of sIL7Rα.

Integrin cytoplasmic tail interactions.

Integrins are heterodimeric cell surface adhesion receptors essential for multicellular life. They connect cells to the extracellular environment and transduce chemical and mechanical signals to and from the cell. Intracellular proteins that bind the integrin cytoplasmic tail regulate integrin engagement of extracellular ligands as well as integrin localization and trafficking. Cytoplasmic integrin-binding proteins also function downstream of integrins, mediating links to the cytoskeleton and to signaling cascades that impact cell motility, growth, and survival. Here, we review key integrin-interacting proteins and their roles in regulating integrin activity, localization, and signaling.

Zasp regulates integrin activation.

Integrins are heterodimeric adhesion receptors that link the extracellular matrix (ECM) to the cytoskeleton. Binding of the scaffold protein, talin, to the cytoplasmic tail of ß-integrin causes a conformational change of the extracellular domains of the integrin heterodimer, thus allowing high-affinity binding of ECM ligands. This essential process is called integrin activation. Here we report that the Z-band alternatively spliced PDZ-motif-containing protein (Zasp) cooperates with talin to activate α5ß1 integrins in mammalian tissue culture and αPS2ßPS integrins in Drosophila. Zasp is a PDZ-LIM-domain-containing protein mutated in human cardiomyopathies previously thought to function primarily in assembly and maintenance of the muscle contractile machinery. Notably, Zasp is the first protein shown to co-activate α5ß1 integrins with talin and appears to do so in a manner distinct from known αIIbß3 integrin co-activators.

Culturally sensitive emergency care for sexual and gender minority youth: A quality improvement initiative.

BACKGROUND: Despite evidence of the impact of provider implicit bias and overt discrimination experienced by sexual and gender minority youth (SGMY), evidence surrounding sexual and gender minority cultural sensitivity training for pediatric emergency health professionals is limited. No targeted training existed to improve the clinical preparedness of healthcare professionals serving SGMY by increasing providers' knowledge and attitudinal awareness in a pediatric emergency department at a large, urban pediatric hospital in the Southeastern United States. METHODS: The Institute for Healthcare Improvement's [15] Model for Improvement informed the project and was completed in four Plan-Do-Study-Act cycles. A cross-sectional, pre-test post-test design was used to gather demographic data, administer the LGBT-DOCSS questionnaire, and collect participant feedback on the training session. The LGBT-DOCSS results were analyzed using an independent samples t-test. INTERVENTIONS: Evidence-based pedagogical strategies were utilized for a 60-minute staff training session. Staff (n = 25) had six opportunities to attend one of the training sessions over a period of 4 months. RESULTS: Self-selection and voluntary participation contributed to recruiting participants who demonstrated high baseline LGBT-DOCSS scores, particularly on the subscales that measure knowledge and attitudinal awareness. After the sessions, participants showed an increase in LGBT-DOCSS scores with a statistically significant increase in the clinical preparedness subscale. CONCLUSIONS: This project was the first at the institution to focus on culturally sensitive emergency care for sexual and gender minority youth. The content was well received by staff, who demonstrated increased clinical preparedness after the training. Implementing the training as a required component of new nurse orientation and onboarding is the next step in creating a safety culture for SGMY in the PED setting.

Chemical genetic strategy identifies histone deacetylase 1 (HDAC1) and HDAC2 as therapeutic targets in sickle cell disease.

The worldwide burden of sickle cell disease is enormous, with over 200,000 infants born with the disease each year in Africa alone. Induction of fetal hemoglobin is a validated strategy to improve symptoms and complications of this disease. The development of targeted therapies has been limited by the absence of discrete druggable targets. We developed a unique bead-based strategy for the identification of inducers of fetal hemoglobin transcripts in primary human erythroid cells. A small-molecule screen of bioactive compounds identified remarkable class-associated activity among histone deacetylase (HDAC) inhibitors. Using a chemical genetic strategy combining focused libraries of biased chemical probes and reverse genetics by RNA interference, we have identified HDAC1 and HDAC2 as molecular targets mediating fetal hemoglobin induction. Our findings suggest the potential of isoform-selective inhibitors of HDAC1 and HDAC2 for the treatment of sickle cell disease.

Thermal comfort and safety of cotton blankets warmed at 130°F and 200°F.

BACKGROUND: In 2009, the ECRI Institute recommended warming cotton blankets in cabinets set at 130°F or less. However, there is limited research to support the use of this cabinet temperature. PURPOSE: To measure skin temperatures and thermal comfort in healthy volunteers before and after application of blankets warmed in cabinets set at 130 and 200°F, respectively, and to determine the time-dependent cooling of cotton blankets after removal from warming cabinets set at the two temperatures. DESIGN: Prospective, comparative, descriptive. METHODS: Participants (n = 20) received one or two blankets warmed in 130 or 200°F cabinets. First, skin temperatures were measured, and thermal comfort reports were obtained at fixed timed intervals. Second, blanket temperatures (n = 10) were measured at fixed intervals after removal from the cabinets. FINDING: No skin temperatures approached levels reported in the literature that cause epidermal damage. Thermal comfort reports supported using blankets from the 200°F cabinet, and blankets lost heat quickly over time. CONCLUSIONS: We recommend warming cotton blankets in cabinets set at 200°F or less to improve thermal comfort without compromising patient safety.

Defining the role of syndecan-4 in mechanotransduction using surface-modification approaches.

The ability of cells to respond to external mechanical stimulation is a complex and robust process involving a diversity of molecular interactions. Although mechanotransduction has been heavily studied, many questions remain regarding the link between physical stimulation and biochemical response. Of significant interest has been the contribution of the transmembrane proteins involved, and integrins in particular, because of their connectivity to both the extracellular matrix and the cytoskeleton. Here, we demonstrate the existence of a mechanically based initiation molecule, syndecan-4. We first demonstrate the ability of syndecan-4 molecules to support cell attachment and spreading without the direct extracellular binding of integrins. We also examine the distribution of focal adhesion-associated proteins through controlling surface interactions of beads with molecular specificity in binding to living cells. Furthermore, after adhering cells to elastomeric membranes via syndecan-4-specific attachments we mechanically strained the cells via our mechanical stimulation and polymer surface chemical modification approach. We found ERK phosphorylation similar to that shown for mechanotransductive response for integrin-based cell attachments through our elastomeric membrane-based approach and optical magnetic twisting cytometry for syndecan-4. Finally, through the use of cytoskeletal disruption agents, this mechanical signaling was shown to be actin cytoskeleton dependent. We believe that these results will be of interest to a wide range of fields, including mechanotransduction, syndecan biology, and cell-material interactions.

Unplanned perioperative hypothermia and agreement between oral, temporal artery, and bladder temperatures in adult major surgery patients.

Accurate body core temperature measurement is essential in perioperative areas to quickly recognize and address abnormal temperatures. The purposes of this prospective, descriptive study were to accurately identify unplanned perioperative hypothermia (UPH) in 64 elective major surgery patients; to describe factors that increased the risk of UPH; to describe active/passive warming measures; to describe thermal comfort in patients with and without UPH; and to compare oral, temporal artery, and bladder temperatures. Based on bladder temperatures, 52% of the patients had UPH in the operating room (OR) and 42% on postanesthesia care unit (PACU) admission. The temporal artery thermometer did not detect any hypothermia. Descriptive data and Bland-Altman plots showed lack of agreement between the temporal artery thermometer readings and those of the oral and bladder thermometers. The patient's thermal comfort report did not accurately reflect hypothermia. Factors found to increase the risk of UPH included older age, BMI lower than 30, and OR ambient temperature lower than 68°F. All but one patient had active warming in the OR; active warming was infrequently used in the PACU. Based on our findings and findings in previous studies, we do not recommend using the temporal artery thermometer in perioperative areas. To prevent UPH, we recommend aggressive use of convective and conductive warming measures in perioperative areas and increasing OR ambient temperatures.

Molecular basis for integrin adhesion receptor binding to p21-activated kinase 4 (PAK4).

Integrin adhesion receptors provide links between extracellular ligands and cytoplasmic signaling. Multiple kinases have been found to directly engage with integrin ß tails, but the molecular basis for these interactions remain unknown. Here, we assess the interaction between the kinase domain of p21-activated kinase 4 (PAK4) and the cytoplasmic tail of integrin ß5. We determine three crystal structures of PAK4-ß5 integrin complexes and identify the PAK-binding site. This is a region in the membrane-proximal half of the ß5 tail and confirmed by site-directed mutagenesis. The ß5 tail engages the kinase substrate-binding groove and positions the non-phosphorylatable integrin residue Glu767 at the phosphoacceptor site. Consistent with this, integrin ß5 is poorly phosphorylated by PAK4, and in keeping with its ability to occlude the substrate-binding site, weakly inhibits kinase activity. These findings demonstrate the molecular basis for ß5 integrin-PAK4 interactions but suggest modifications in understanding the potential cellular role of this interaction.

Transgenic Drosophila lines for LexA-dependent gene and growth regulation.

Conditional expression of short hairpin RNAs with binary genetic systems is an indispensable tool for studying gene function. Addressing mechanisms underlying cell-cell communication in vivo benefits from simultaneous use of 2 independent gene expression systems. To complement the abundance of existing Gal4/UAS-based resources in Drosophila, we and others have developed LexA/LexAop-based genetic tools. Here, we describe experimental and pedagogical advances that promote the efficient conversion of Drosophila Gal4 lines to LexA lines, and the generation of LexAop-short hairpin RNA lines to suppress gene function. We developed a CRISPR/Cas9-based knock-in system to replace Gal4 coding sequences with LexA, and a LexAop-based short hairpin RNA expression vector to achieve short hairpin RNA-mediated gene silencing. We demonstrate the use of these approaches to achieve targeted genetic loss-of-function in multiple tissues. We also detail our development of secondary school curricula that enable students to create transgenic flies, thereby magnifying the production of well-characterized LexA/LexAop lines for the scientific community. The genetic tools and teaching methods presented here provide LexA/LexAop resources that complement existing resources to study intercellular communication coordinating metazoan physiology and development.

A novel HDAC inhibitor with a hydroxy-pyrimidine scaffold.

Histone deacetylases (HDACs) are enzymes involved in many important biological functions. They have been linked to a variety of cancers, psychiatric disorders, and other diseases. Since small molecules can serve as probes to study the relevant biological roles of HDACs, novel scaffolds are necessary to develop more efficient, selective drug candidates. Screening libraries of molecules may yield structurally diverse probes that bind these enzymes and modulate their functions in cells. Here we report a small molecule with a novel hydroxy-pyrimidine scaffold that inhibits multiple HDAC enzymes and modulates acetylation levels in cells. Analogs were synthesized in an effort to evaluate structure-activity relationships.

TRB3 is stimulated in diabetic kidneys, regulated by the ER stress marker CHOP, and is a suppressor of podocyte MCP-1.

The prevalence of diabetic nephropathy continues to rise, highlighting the importance of investigating and discovering novel treatment strategies. TRB3 is a kinase-like molecule that modifies cellular survival and metabolism and interferes with signal transduction pathways. Herein, we report that TRB3 expression is increased in the kidneys of type 1 and type 2 diabetic mice. TRB3 is expressed in conditionally immortalized podocytes; however, it is not stimulated by elevated glucose. The diabetic milieu is associated with increased oxidative stress and circulating free fatty acids (FFA). We show that reactive oxygen species (ROS) such as H(2)O(2) and superoxide anion (via the xanthine/xanthine oxidase reaction) as well as the FFA palmitate augment TRB3 expression in podocytes. C/EBP homologous protein (CHOP) is a transcription factor that is associated with the endoplasmic reticulum stress response. CHOP expression increases in diabetic mouse kidneys and in podocytes treated with ROS and FFA. In podocytes, transfection of CHOP increases TRB3 expression, and ROS augment recruitment of CHOP to the proximal TRB3 promoter. MCP-1/CCL2 is a chemokine that contributes to the inflammatory injury associated with diabetic nephropathy. In these studies, we demonstrate that TRB3 can inhibit basal and stimulated podocyte production of MCP-1. In summary, enhanced ROS and/or FFA associated with the diabetic milieu induce podocyte CHOP and TRB3 expression. Because TRB3 inhibits MCP-1, manipulation of TRB3 expression could provide a novel therapeutic approach in diabetic kidney disease.

The Relationship Between Critical Care Work Environment and Professional Quality of Life.

BACKGROUND: Professional quality of life is the quality a person feels in relation to work. For critical care nurses, it is composed of compassion satisfaction and compassion fatigue. Professional quality of life is affected by work environment. The American Association of Critical-Care Nurses (AACN) has identified 6 standards for a healthy work environment. OBJECTIVE: To explore which of the AACN healthy work environment standards have the strongest impact on professional quality of life in critical care nurses. METHODS: In an exploratory, cross-sectional survey of nurses working in 4 adult critical care units of a single health care facility, professional quality of life was assessed using the Professional Quality of Life Scale (ProQOL), and work environment was evaluated using the AACN Healthy Work Environment Assessment Tool. RESULTS: Participants reported compassion satisfaction and burnout levels as average and secondary traumatic stress levels as high. The composite average for all 6 AACN healthy work environment standards was good. A multiple regression analysis revealed true collaboration, effective decision-making, and authentic leadership as significant predictors of compassion satisfaction. Authentic leadership was the only predictor of burnout. Appropriate staffing, meaningful recognition, and authentic leadership were predictors of secondary traumatic stress. CONCLUSION: Authentic leadership is the strongest predictor of compassion satisfaction, burnout, and secondary traumatic stress. Therefore, improving leadership should be a priority in intensive care units seeking to improve nurses' professional quality of life.

Barriers and Facilitators of Breastfeeding Reported by Postpartum Women in Methadone Maintenance Therapy.

INTRODUCTION: This study utilized a cross-sectional qualitative and quantitative interview-based survey to capture the infant feeding practices and barriers to exclusive breastfeeding for women in methadone maintenance therapy. Participants were recruited from an opioid dependence treatment center in an urban setting in the Southeastern United States. MATERIALS AND METHODS: A convenience sample of women in treatment (n = 30) were interviewed using an adapted instrument designed to capture decisions and intentions to formula feed or breastfeed; support from friends and family; hospital experience; support from healthcare personnel; and maternal knowledge of breastfeeding while taking methadone. RESULTS: The majority of women in the sample initiated breastfeeding, but only 10% continued for >1 month. Challenges related to infant hospital stay posed a significant barrier. Two-thirds of infants remained hospitalized after the mother was discharged. Out of the 24 women who initiated breastfeeding, 11 reported that they discontinued because of issues related to infant's neonatal intensive care unit (NICU) stay. Eleven women reported that their healthcare providers did not discuss breastfeeding with them. Women who were encouraged to breastfeed by healthcare staff were more likely to breastfeed for longer durations. CONCLUSIONS: Women in treatment for opioid dependence both desire and attempt to establish breastfeeding, but encounter significant challenges, including long NICU stays and lack of support and education, that compromise their success. These findings should inform the development of future programs or interventions geared toward increasing breastfeeding initiation, support, and duration among women who give birth to babies while in treatment for opioid addiction.