ELF4 is critical for induction of type I interferon and the host antiviral response.

Induction of type I interferon is a central event of innate immunity, essential for host defense. Here we report that the transcription factor ELF4 is induced by type I interferon and upregulates interferon expression in a feed-forward loop. ELF4 deficiency leads to reduced interferon production, resulting in enhanced susceptibility to West Nile virus encephalitis in mice. After viral infection, ELF4 is recruited by STING, interacts with and is activated by the MAVS-TBK1 complex, and translocates into the nucleus to bind interferon promoters. Cooperative binding with ELF4 increases the binding affinity of interferon regulatory factors IRF3 and IRF7, which is mediated by EICE elements. Thus, in addition to identifying a regulator of innate immune signaling, we uncovered a role for EICE elements in interferon transactivation.

In-hospital end-of-life care: an appreciative analysis of bereaved family feedback.

Experience feedback data is increasingly recognised as being helpful in improving healthcare services, and in meeting patient and family needs. This end-of-life care project, based on the principles of appreciative inquiry, sought to learn from the experiences of bereaved people whose relative had died in an acute hospital setting. Informal feedback, offered during a routine telephone call, was thematically analysed and interpreted in an appreciative manner. Confirmatory representations of caring practices and behaviours were identified, categorised and disseminated in a way that enabled staff to come to know and understand end-of-life care at its best, rather than as a set of problematised events. The findings served as a benchmark for individuals and teams to assess and progress their practice reflectively. The authors conclude that staff receptiveness to informal bereaved family feedback may be enhanced by focusing on the positive qualities of end-of-life care within existing practices.

IRF3 Signaling within the Mouse Stroma Influences Sepsis Pathogenesis.

IFN regulatory factor 3 (IRF3) is a transcription factor that is activated by multiple pattern-recognition receptors. We demonstrated previously that IRF3 plays a detrimental role in a severe mouse model of sepsis, induced by cecal ligation and puncture. In this study, we found that IRF3-knockout (KO) mice were greatly protected from sepsis in a clinically relevant version of the cecal ligation and puncture model incorporating crystalloid fluids and antibiotics, exhibiting improved survival, reduced disease score, lower levels of serum cytokines, and improved phagocytic function relative to wild-type (WT) mice. Computational modeling revealed that the overall complexity of the systemic inflammatory/immune network was similar in IRF3-KO versus WT septic mice, although the tempo of connectivity differed. Furthermore, the mediators driving the network differed: TNF-α, IL-1ß, and IL-6 predominated in WT mice, whereas MCP-1 and IL-6 predominated in IRF3-KO mice. Network analysis also suggested differential IL-6-related inflammatory programs in WT versus IRF3-KO mice. We created bone marrow chimeras to test the role of IRF3 within leukocytes versus stroma. Surprisingly, chimeras with IRF3-KO bone marrow showed little protection from sepsis, whereas chimeras with IRF3-KO stroma showed a substantial degree of protection. We found that WT and IRF3-KO macrophages had a similar capacity to produce IL-6 and phagocytose bacteria in vitro. Adoptive transfer experiments demonstrated that the genotype of the host environment affected the capacity of monocytes to produce IL-6 during sepsis. Thus, IRF3 acts principally within the stromal compartment to exacerbate sepsis pathogenesis via differential impacts on IL-6-related inflammatory programs.

Family experiences of in-hospital end-of-life care for adults: A systematic review of qualitative evidence.

AIM: To systematically identify, appraise, aggregate and synthesise qualitative evidence on family members' experiences of end-of-life care (EoLC) in acute hospitals. METHODS: A systematic review and qualitative evidence synthesis based on the Joanna Briggs Institute methodology. Primary research, published 2014 onwards was identified using a sequential strategy of electronic and hand searches. Six databases (CINAHL, Medline, Embase, EMCare, PsycINFO, BNI) were systematically searched. Studies that met pre-determined inclusion/exclusion criteria were uniformly appraised using the Critical Appraisal Skills Programme checklist for qualitative research, and synthesised using a meta-aggregative approach. The ENTREQ statement was used as a checklist for reporting the review. RESULTS: Sixteen studies of European, Australasian and North American origin formed the review. The quality of each study was considered very good in view of a 'yes' response to most screening questions. Extracted findings were assembled into 12 categories, and five synthesised findings: Understanding of approaching end of life; essential care at the end of life; interpersonal interactions; environment of care; patient and family care after death. CONCLUSION: Enabling and improving peoples' experience of EoLC must remain part of the vision and mission of hospital organisations. Consideration must be given to the fulfilment of family needs and apparent hallmarks of quality care that appear to influence experiential outcomes. RELEVANCE TO CLINICAL PRACTICE: This review of qualitative research represents the first-stage development of a family-reported experience measure for adult EoLC in the hospital setting. The synthesised findings provide a Western perspective of care practices and environmental factors that are perceived to impact the quality of the care experience. Collectively, the review findings serve as a guide for evidence-informed practice, quality improvement, service evaluation and future research. A developed understanding of the families' subjective reflections creates reciprocal opportunity to transform experiential insights into practical strategies for professional growth and practice development.

mPR-Specific Actions Influence Maintenance of the Blood-Brain Barrier (BBB).

Cerebral cavernous malformations (CCMs) are characterized by abnormally dilated intracranial microvascular sinusoids that result in increased susceptibility to hemorrhagic stroke. It has been demonstrated that three CCM proteins (CCM1, CCM2, and CCM3) form the CCM signaling complex (CSC) to mediate angiogenic signaling. Disruption of the CSC will result in hemorrhagic CCMs, a consequence of compromised blood-brain barrier (BBB) integrity. Due to their characteristically incomplete penetrance, the majority of CCM mutation carriers (presumed CCM patients) are largely asymptomatic, but when symptoms occur, the disease has typically reached a clinical stage of focal hemorrhage with irreversible brain damage. We recently reported that the CSC couples both classic (nuclear; nPRs) and nonclassic (membrane; mPRs) progesterone (PRG)-receptors-mediated signaling within the CSC-mPRs-PRG (CmP) signaling network in nPR(-) breast cancer cells. In this report, we demonstrate that depletion of any of the three CCM genes or treatment with mPR-specific PRG actions (PRG/mifepristone) results in the disruption of the CmP signaling network, leading to increased permeability in the nPR(-) endothelial cells (ECs) monolayer in vitro. Finally, utilizing our in vivo hemizygous Ccm mutant mice models, we demonstrate that depletion of any of the three CCM genes, in combination with mPR-specific PRG actions, is also capable of leading to defective homeostasis of PRG in vivo and subsequent BBB disruption, allowing us to identify a specific panel of etiological blood biomarkers associated with BBB disruption. To our knowledge, this is the first report detailing the etiology to predict the occurrence of a disrupted BBB, an indication of early hemorrhagic events.

The circadian clock controls toll-like receptor 9-mediated innate and adaptive immunity.

Circadian rhythms refer to biologic processes that oscillate with a period of ~24 hr. These rhythms are sustained by a molecular clock and provide a temporal matrix that ensures the coordination of homeostatic processes with the periodicity of environmental challenges. We demonstrate the circadian molecular clock controls the expression and function of Toll-like receptor 9 (TLR9). In a vaccination model using TLR9 ligand as adjuvant, mice immunized at the time of enhanced TLR9 responsiveness presented weeks later with an improved adaptive immune response. In a TLR9-dependent mouse model of sepsis, we found that disease severity was dependent on the timing of sepsis induction, coinciding with the daily changes in TLR9 expression and function. These findings unveil a direct molecular link between the circadian and innate immune systems with important implications for immunoprophylaxis and immunotherapy.

Circadian Rhythms Influence the Severity of Sepsis in Mice via a TLR2-Dependent, Leukocyte-Intrinsic Mechanism.

Circadian rhythms coordinate an organism's activities and biological processes to the optimal time in the 24-h daylight cycle. We previously demonstrated that male C57BL/6 mice develop sepsis more rapidly when the disease is induced in the nighttime versus the daytime. In this report, we elucidate the mechanism of this diurnal difference. Sepsis was induced via cecal ligation and puncture (CLP) at zeitgeber time (ZT)-19 (2 am) or ZT-7 (2 pm). Like the males used in our prior study, female C57BL/6 mice had a worse outcome when CLP was induced at ZT-19 versus ZT-7, and these effects persisted when we pooled the data from both sexes. In contrast, mice with a mutated Period 2 (Per2) gene had a similar outcome when CLP was induced at ZT-19 versus ZT-7. Bone marrow chimeras reconstituted with C57BL/6 immune cells exhibited a worse outcome when sepsis was induced at ZT-19 versus ZT-7, whereas chimeras with Per2-mutated immune cells did not. Next, murine macrophages were subjected to serum shock to synchronize circadian rhythms and exposed to bacteria cultured from the mouse cecum at 4-h intervals for 48 h. We observed that IL-6 production oscillated with a 24-h period in C57BL/6 cells exposed to cecal bacteria. Interestingly, we observed a similar pattern when cells were exposed to the TLR2 agonist lipoteichoic acid. Furthermore, TLR2-knockout mice exhibited a similar sepsis phenotype when CLP was induced at ZT-19 versus ZT-7. Together, these data suggest that circadian rhythms in immune cells mediate diurnal variations in murine sepsis severity via a TLR2-dependent mechanism.

Family bereavement: A case study of controlled organ donation after circulatory death.

Deceased organ donation represents a major source of organs for human transplantation practice. In the United Kingdom, as well as other parts of the world, donation after circulatory death accounts for a proportion of all deceased organ donors. Organ and tissue donation emotively takes place in the context of dying, death and bereavement, yet little is known about the family experience of donation after circulatory death. This paper presents a case study of the phenomenon of controlled donation after circulatory death in intensive care. We present a critical analysis of care processes through the lens of a British donor family who participated in a national study of organ and tissue donation. Anonymized family quotes are applied to illustrate specific case issues, and with reference to relevant national guidance and international research. The case portrayed intimate details of the moment in time when the family experienced the potential for controlled donation after circulatory death, factors that appeared to influence family consent and the perceived expectations and outcomes arising from the donation decision. Case analysis demonstrated local compliance with best practice guidance and compassionate end-of-life care while supporting organ retrieval. Caring for the grieving family of potential organ donors requires sensitivity and skill. Of importance is a sound professional knowledge and understanding of the clinical care pathway, together with effective teamwork, optimal communication, family and staff support. Further research is required to determine the impact of controlled donation after circulatory death on family grief and bereavement.

Increased Levels of Macrophage Inflammatory Proteins Result in Resistance to R5-Tropic HIV-1 in a Subset of Elite Controllers.

UNLABELLED: Elite controllers (ECs) are a rare group of HIV seropositive individuals who are able to control viral replication without antiretroviral therapy. The mechanisms responsible for this phenotype, however, have not been fully elucidated. In this study, we examined CD4(+) T cell resistance to HIV in a cohort of elite controllers and explored transcriptional signatures associated with cellular resistance. We demonstrate that a subgroup of elite controllers possess CD4(+) T cells that are specifically resistant to R5-tropic HIV while remaining fully susceptible to X4-tropic and vesicular stomatitis virus G (VSV-G)-pseudotyped viruses. Transcriptome analysis revealed 17 genes that were differentially regulated in resistant elite controllers relative to healthy controls. Notably, the genes encoding macrophage inflammatory protein 1α (MIP-1α), CCL3 and CCL3L1, were found to be upregulated. The MIP-1α, MIP-1ß, and RANTES chemokines are natural ligands of CCR5 and are known to interfere with HIV replication. For three elite controllers, we observed increased production of MIP-1α and/or MIP-1ß at the protein level. The supernatant from resistant EC cells contained MIP-1α and MIP-1ß and was sufficient to confer R5-tropic resistance to susceptible CD4(+) T cells. Additionally, this effect was reversed by using inhibitory anti-MIP antibodies. These results suggest that the T cells of these particular elite controllers may be naturally resistant to HIV infection by blocking R5-tropic viral entry. IMPORTANCE: HIV is a pandemic health problem, and the majority of seropositive individuals will eventually progress to AIDS unless antiretroviral therapy (ART) is administered. However, rare patients, termed elite controllers, have a natural ability to control HIV infection in the absence of ART, but the mechanisms by which they achieve this phenotype have not been fully explored. This paper identifies one mechanism that may contribute to this natural resistance: some elite controllers have CD4(+) T cells that produce high levels of MIP chemokines, which block R5-tropic HIV entry. This mechanism could potentially be exploited to achieve a therapeutic effect in other HIV-seropositive individuals.

Incidence, aetiology and outcome of pleural empyema and parapneumonic effusion from 1998 to 2012 in a population of New Zealand children.

AIM: To document rising incidence rates of childhood empyema and parapneumonic effusion (PPE) in South Auckland, New Zealand between 1998 and 2012; to compare epidemiology, pathogens and outcomes of children with empyema and PPE; and to ascertain whether primary care antibiotic prescribing, delayed presentation, or bacterial epidemiology might account for the rising incident rates. METHODS: Children aged 0 to14 years hospitalised with pleural empyema or PPE were retrospectively identified. Empyema was defined by ultrasound and pleural tap criteria. PPE was defined as the presence of pleural fluid on chest xray not fulfilling empyema criteria. Epidemiology, clinical features, microbiology and outcomes of empyema and PPE were compared and incidence rates analysed. RESULTS: Of 184 cases identified, 104 met the criteria for empyema. Empyema incidence increased from 1 per 100 000 children aged 0 to 14 years in 1998 to 10 per 100 000 in 2012, with a peak incidence of 13 per 100 000 in 2009. Staphylococcus aureus was most frequently detected (n=38), followed by Streptococcus pneumoniae (n=31). Cases of S. aureus empyema increased 4 fold over the 15 years. Dominant S. pneumoniae serotypes were 1 and 14. Thirty-five percent of empyema and 53% of PPE cases received pre-hospital antibiotics. Children who received pre-hospital antibiotics were more than 40% less likely to require surgical intervention than those not pre-treated. CONCLUSIONS: Childhood empyema incidence has increased markedly in South Auckland. Paediatric S. aureus empyema is becoming increasingly common in South Auckland. Pre-hospital antibiotic prescribing may mitigate the need for surgical intervention in our population.

Immunity's fourth dimension: approaching the circadian-immune connection.

The circadian system ensures the generation and maintenance of self-sustained ~24-h rhythms in physiology that are linked to internal and environmental changes. In mammals, daily variations in light intensity and other cues are integrated by a hypothalamic master clock that conveys circadian information to peripheral molecular clocks that orchestrate physiology. Multiple immune parameters also vary throughout the day and disruption of circadian homeostasis is associated with immune-related disease. Here, we discuss the molecular links between the circadian and immune systems and examine their outputs and disease implications. Understanding the mechanisms that underlie circadian-immune crosstalk may prove valuable for devising novel prophylactic and therapeutic interventions.

Assessment of single-bolus contrast administration technique using hybrid dual-source ECG-gated thoracic and dual-source non-ECG-gated high-pitch abdominopelvic CT acquisitions for procedural planning before transcatheter aortic valve replacement.

PURPOSE: This study aimed to assess vascular contrast opacification and homogeneity using single-bolus contrast administration with hybrid thoracic and abdominopelvic computed tomographic angiography in patients with severe aortic valve stenosis. MATERIALS AND METHODS: Combination electrocardiogram-gated thoracic and dual-source, high-pitch abdominopelvic computed tomographic angiography examinations of 50 patients with severe aortic stenosis between December 2013 and March 2014 were reviewed. Contrast administration was individualized to patient-specific physiology. Image analysis of vascular opacification was obtained and interdependencies of vascular contrast and homogeneity of contrast distribution were assessed. RESULTS: The mean volume of contrast administered was 106 ± 11.7 mL. Mean attenuation was 371 ± 90.7 Hounsfield units (HU) in the thoracic aorta and 388 ± 95.9 HU in the abdominal aorta. Homogeneous opacification was obtained throughout with coefficient of variation of 11%. CONCLUSIONS: Procedural planning for transcatheter aortic valve replacement can be achieved using a single-injection bolus contrast protocol in combination with a 2-part multidetector computed tomographic image acquisition technique with optimal opacification of major arterial structures.

MKK3 regulates mitochondrial biogenesis and mitophagy in sepsis-induced lung injury.

Sepsis is a systemic inflammatory response to infection and a major cause of death worldwide. Because specific therapies to treat sepsis are limited, and underlying pathogenesis is unclear, current medical care remains purely supportive. Therefore targeted therapies to treat sepsis need to be developed. Although an important mediator of sepsis is thought to be mitochondrial dysfunction, the underlying molecular mechanism is unclear. Modulation of mitochondrial processes may be an effective therapeutic strategy in sepsis. Here, we investigated the role of the kinase MKK3 in regulation of mitochondrial function in sepsis. Using clinically relevant animal models, we examined mitochondrial function in primary mouse lung endothelial cells exposed to LPS. MKK3 deficiency reduces lethality of sepsis in mice and by lowering levels of lung and mitochondrial injury as well as reactive oxygen species. Furthermore, MKK3 deficiency appeared to simultaneously increase mitochondrial biogenesis and mitophagy through the actions of Sirt1, Pink1, and Parkin. This led to a more robust mitochondrial network, which we propose provides protection against sepsis. We also detected higher MKK3 activation in isolated peripheral blood mononuclear cells from septic patients compared with nonseptic controls. Our findings demonstrate a critical role for mitochondria in the pathogenesis of sepsis that involves a previously unrecognized function of MKK3 in mitochondrial quality control. This mitochondrial pathway may help reveal new diagnostic markers and therapeutic targets against sepsis.

Emergency care staff experiences of lay presence during adult cardiopulmonary resuscitation: a phenomenological study.

BACKGROUND: Public support in favour of family presence during an adult cardiopulmonary resuscitation (CPR) attempt is a contentious issue among providers of emergency care. Researchers have mostly relied on attitudinal surveys to elicit staff views, leaving the life-world of those who have experienced this phenomenon, largely unexplored. OBJECTIVE: To explore the lived experience of lay presence during an adult CPR attempt in primary (out-of-hospital) and secondary (inhospital) environments of care. DESIGN: Hermeneutical phenomenological study. METHODS: Semistructured, face-to-face interviews with 8 ambulance staff and 12 registered nurses. The interviews were audio-recorded and subjected to thematic analysis. RESULTS: Participants provided insight into situations where lay presence during adult CPR came about either spontaneously or as a planned event. Their accounts portrayed a mixture of benefits and concerns. Familiarity of working in the presence of lay people, practical experience in emergency care and personal confidence were important antecedents. Divergent practices within and across the contexts of care were revealed. The concept of exposure emerged as the essence of this phenomenon. Overall, the study findings serve to challenge some of the previously reported attitudes and opinions of emergency care staff. CONCLUSIONS: Improved intraprofessional and interprofessional collaboration is essential to overcoming the barriers associated with lay presence during adult CPR. The future of this practice is dependent on initiatives that seek to bring about attitudinal change. Priority should be given to further exploring this phenomenon in the context of patient and family centred end-of-life care.

Intensive care nurses' experiences of providing end-of-life care after treatment withdrawal: a qualitative study.

AIMS AND OBJECTIVES: To explore the experiences of intensive care nurses who provided end-of-life care to adult patients and their families after a decision had been taken to withdraw treatment. BACKGROUND: End-of-life care following treatment withdrawal is a common phenomenon in intensive care. Less is known about nurses' experiences of providing care for the dying patient and their family in this context, when compared to specialist palliative care. DESIGN: Descriptive exploratory qualitative study. METHODS: A purposive sample of 13 intensive care nurses participated in a semistructured face-to-face interview. Transcribed data were analysed using the principles of interpretative phenomenological analysis. RESULTS: The essence of nurses' experiences of providing end-of-life care after the withdrawal of treatment was interpreted as 'doing the best to facilitate a comfortable and dignified death'. Four master themes included the following: caring for the dying patient and their family; providing and encouraging presence; reconnecting the patient and family; and dealing with emotions and ambiguity. Uncertainties were evident on processes and actions involved in treatment withdrawal, how to reconnect patients and their family effectively and how to reduce the technological environment. CONCLUSIONS: Providing end-of-life care after a decision has been taken to withdraw treatment was a common aspect of intensive care. It was evident that nurses were doing their utmost to support patients and families at the end of life, despite the multiple challenges they faced. RELEVANCE TO CLINICAL PRACTICE: The interpretive findings from this study should assist intensive care unit nurses to better understand and develop their role in providing high-quality end-of-life care after treatment withdrawal. Practice guidelines should be developed to reduce ambiguity and support the delivery of high-quality care for adults as they approach the final stages of life in intensive care units.