Unravelling the anti-inflammatory mechanisms of dietary fatty acids

Polyunsaturated fatty acids (PUFA) have been shown to modulate immune responses and have therapeutic effects in inflammatory disorders. The specific mechanisms of their actions have yet to be defined. The objective of this work was to elucidate such mechanisms. Macrophages are a key component of the innate response, which express toll-like receptors (TLRs). Ligation of TLR4, by its ligand lipopolysaccharide (LPS), results in macrophage activation. This study demonstrates that the n-6 derivative, conjugated linoleic acid (CLA) and n-3 PUFA, DHA and EPA differentially modulate the response of macrophages to LPS. Specifically, phagocytosis was enhanced by CLA and suppressed by n-3 PUFA and these PUFA suppressed TNFα, IL-6 and enhanced IL-10 production, rendering the macrophage less inflammatory. PUFA also suppressed macrophage migration in response to LPS and inhibited production of chemokines. Furthermore, CLA inhibited activation of the TLR4 downstream transcription factors NF-κB and IRF3, while n-3 PUFA, DHA and EPA solely inhibited NF-κB. Further investigation revealed that PUFA selectively regulate the expression of TLR4 and its associated molecule CD14 in response to LPS, but had no effect on LPS binding to TLR4. The exact mechanism of the effects of PUFA on CD14 was elucidated by examining lipid raft 'microdomains', the location where the receptor complex clusters upon activation. We found that treatment of macrophages with CLA reduced the incorporation of CD14 into lipid rafts following activation with LPS. We then examined endocytosis of TLR4 given the role of CD14 in this process, and we found that it was suppressed by CLA. This study therefore reveals a novel mechanism whereby CLA exerts its antiinflammatory effects. This involves suppression of CD14, the subsequent suppression of TLR4 endocytosis culminating in decreased IRF3 activation

Implicit and explicit pedagogical practices related to sociocultural issues and social justice in physical education teacher education programs

Background: For many years, scholars in PETE have argued for the importance of educating pre-service teachers (PSTs) about equality (e.g., Evans 1990), sociocultural perspectives and issues (e.g., Cliff, Wright and Clarke, 2009; Author 2014) and critical pedagogy (e.g., Fernandez-Balboa 1997; Philpot 2015). Despite this advocacy, we would argue that there are significant differences in how faculty teach about sociocultural issues, and for, social justice. The pedagogical actions through which Physical Education Teacher Educators (PETEs) do this work is the focus of this paper. Purpose: We investigated the pedagogical approaches and strategies used by PETE faculty to address and educate PSTs about social justice and sociocultural issues related to gender, race, sexuality, (dis)ability, socioeconomic status and religion in their individual PETE programs. In this study, we draw on transformational pedagogy (Ukpokodu 2009; Ovens 2017) as a framework for theorizing the data. Through this study, we highlight the pedagogical practices espoused as those that engender transformative learning. Data collection and analysis: Data for this interpretive qualitative research study was collected primarily through in-depth semi-structured interviews with over 70 PETEs who work in 48 PETE programs across Australia, Canada, England, Ireland New Zealand, Sweden, and the United States. Furthermore, an informational survey was used to gather demographic data of the participants. The participants, all current PETEs, had a wide range of professional experiences, which included the length of time in the profession, the type of institution employed, educational backgrounds and courses taught. Data analysis was completed using the processes of content analysis and the constant comparative method (Corbin and Strauss 2008). Findings: Three major themes represent the findings. In the first theme, ‘Intentional and Explicit Pedagogies’ we provide descriptions of the approaches and strategies used by PETEs in this study that were planned in advance of the learning experiences. In the second theme, ‘Teachable Moments’ we provide examples of how PETEs utilized ‘teachable moments’ in implicit and explicit ways to educate PSTs about sociocultural issues. The third theme, ‘Resistance and Constraints’ captures the individual challenges PETE faculty faced within their courses if, and when, they teach for equity and social justice. The findings suggest that social justice struggles to find an explicit presence within many PETE programs and that educating PSTs about sociocultural issues and social justice is lacking in many PETE programs

Membrane vesicles from Pseudomonas aeruginosa activate the non-canonical inflammasome through caspase-5 in human monocytes

Outer membrane vesicles (OMVs) are constitutively produced by Gram-negative bacteria both in vivo and in vitro. These lipid-bound structures carry a range of immunogenic components derived from the parent cell, which are transported into host target cells and activate the innate immune system. Recent advances in the field have shed light on some of the multifaceted roles of OMVs in host-pathogen interactions. In this study, we investigated the ability of OMVs from two clinically important pathogens, Pseudomonas aeruginosa and Helicobacter pylori, to activate canonical and non-canonical inflammasomes. P.\ua0aeruginosa OMVs induced inflammasome activation in mouse macrophages, as evidenced by "speck" formation, as well as the cleavage and secretion of interleukin-1β and caspase-1. These responses were independent of AIM2 and NLRC4 canonical inflammasomes, but dependent on the non-canonical caspase-11 pathway. Moreover, P.\ua0aeruginosa OMVs alone were able to activate the inflammasome in a TLR-dependent manner, without requiring an exogenous priming signal. In contrast, H.\ua0pylori OMVs were not able to induce inflammasome activation in macrophages. Using CRISPR/Cas9 knockout THP-1 cells lacking the human caspase-11 homologs, caspase-4 and -5, we demonstrated that caspase-5 but not caspase-4 is required for inflammasome activation by P. aeruginosa OMVs in human monocytes. In contrast, free P.\ua0aeruginosa LPS transfected into cells induced inflammasome responses via caspase-4. This suggests that caspase-4 and caspase-5 differentially recognize LPS depending on its physical form or route of delivery into the cell. These findings have relevance to Gram-negative infections in humans and the use of OMVs as novel vaccines. This article is protected by copyright. All rights reserved

Child Health Partnerships: a review of program characteristics, outcomes and their relationship

<p>Abstract</p> <p>Background</p> <p>Novel approaches are increasingly employed to address the social determinants of health of children world-wide. Such approaches have included complex social programs involving multiple stakeholders from different sectors jointly working together (hereafter Child Health Partnerships). Previous reviews have questioned whether these programs have led to significant improvements in child health and related outcomes. We aim to provide definitive answers to this question as well as identifying the characteristics of successful partnerships.</p> <p>Methods</p> <p>A comprehensive literature search identified 11 major Child Health Partnerships in four comparable developed countries. A critical review is focused on various aspects of these including their target groups, program mechanics and outcomes.</p> <p>Results and Conclusions</p> <p>There was evidence of success in several major areas from the formation of effective joint operations of partners in different partnership models to improvement in both child wellbeing and parenting. There is emerging evidence that Child Health Partnerships are cost-effective. Population characteristics and local contexts need to be taken into account in the introduction and implementation of these programs.</p

The masked demos: Associational anonymity and democratic practice

The increased use of anonymous digital platforms raises substantive concerns about accountability in digital spaces. However, contemporary evaluations of anonymity focus too narrowly on its protective function: its ability to protect a diversity of speakers and ideas. Drawing on two examples of anonymous political engagements – Publius’s writing of the Federalist Papers and college students’ use of the social media platform Yik Yak – we develop an account of anonymity’s associational function: the processes by which people generate and negotiate collective identities, discussions, and actions in wider publics. As we argue, anonymity’s associational function can (1) generate conditions under which individuals develop collective interests and identities to foster collective action, and (2) enable novel interactions between these individuals and communities and the larger publics of which they are part. We conclude with a discussion of how attention to associational anonymity can contribute to a more nuanced account of democracy in practice

Mycobacterial PIMs Inhibit Host Inflammatory Responses through CD14-Dependent and CD14-Independent Mechanisms

Mycobacteria develop strategies to evade the host immune system. Among them, mycobacterial LAM or PIMs inhibit the expression of pro-inflammatory cytokines by activated macrophages. Here, using synthetic PIM analogues, we analyzed the mode of action of PIM anti-inflammatory effects. Synthetic PIM1 isomer and PIM2 mimetic potently inhibit TNF and IL-12 p40 expression induced by TLR2 or TLR4 pathways, but not by TLR9, in murine macrophages. We show inhibition of LPS binding to TLR4/MD2/CD14 expressing HEK cells by PIM1 and PIM2 analogues. More specifically, the binding of LPS to CD14 was inhibited by PIM1 and PIM2 analogues. CD14 was dispensable for PIM1 and PIM2 analogues functional inhibition of TLR2 agonists induced TNF, as shown in CD14-deficient macrophages. The use of rough-LPS, that stimulates TLR4 pathway independently of CD14, allowed to discriminate between CD14-dependent and CD14-independent anti-inflammatory effects of PIMs on LPS-induced macrophage responses. PIM1 and PIM2 analogues inhibited LPS-induced TNF release by a CD14-dependent pathway, while IL-12 p40 inhibition was CD14-independent, suggesting that PIMs have multifold inhibitory effects on the TLR4 signalling pathway

A Role for TLR4 in Clostridium difficile Infection and the Recognition of Surface Layer Proteins

Clostridium difficile is the etiological agent of antibiotic-associated diarrhoea (AAD) and pseudomembranous colitis in humans. The role of the surface layer proteins (SLPs) in this disease has not yet been fully explored. The aim of this study was to investigate a role for SLPs in the recognition of C. difficile and the subsequent activation of the immune system. Bone marrow derived dendritic cells (DCs) exposed to SLPs were assessed for production of inflammatory cytokines, expression of cell surface markers and their ability to generate T helper (Th) cell responses. DCs isolated from C3H/HeN and C3H/HeJ mice were used in order to examine whether SLPs are recognised by TLR4. The role of TLR4 in infection was examined in TLR4-deficient mice. SLPs induced maturation of DCs characterised by production of IL-12, TNFα and IL-10 and expression of MHC class II, CD40, CD80 and CD86. Furthermore, SLP-activated DCs generated Th cells producing IFNγ and IL-17. SLPs were unable to activate DCs isolated from TLR4-mutant C3H/HeJ mice and failed to induce a subsequent Th cell response. TLR4−/− and Myd88−/−, but not TRIF−/− mice were more susceptible than wild-type mice to C. difficile infection. Furthermore, SLPs activated NFκB, but not IRF3, downstream of TLR4. Our results indicate that SLPs isolated from C. difficile can activate innate and adaptive immunity and that these effects are mediated by TLR4, with TLR4 having a functional role in experimental C. difficile infection. This suggests an important role for SLPs in the recognition of C. difficile by the immune system

Mitochondrial arginase-2 is essential for IL-10 metabolic reprogramming of inflammatory macrophages.

Mitochondria are important regulators of macrophage polarisation. Here, we show that arginase-2 (Arg2) is a microRNA-155 (miR-155) and interleukin-10 (IL-10) regulated protein localized at the mitochondria in inflammatory macrophages, and is critical for IL-10-induced modulation of mitochondrial dynamics and oxidative respiration. Mechanistically, the catalytic activity and presence of Arg2 at the mitochondria is crucial for oxidative phosphorylation. We further show that Arg2 mediates this process by increasing the activity of complex II (succinate dehydrogenase). Moreover, Arg2 is essential for IL-10-mediated downregulation of the inflammatory mediators succinate, hypoxia inducible factor 1α (HIF-1α) and IL-1β in vitro. Accordingly, HIF-1α and IL-1β are highly expressed in an LPS-induced in vivo model of acute inflammation using Arg2-/- mice. These findings shed light on a new arm of IL-10-mediated metabolic regulation, working to resolve the inflammatory status of the cell

AI is a viable alternative to high throughput screening: a 318-target study

: High throughput screening (HTS) is routinely used to identify bioactive small molecules. This requires physical compounds, which limits coverage of accessible chemical space. Computational approaches combined with vast on-demand chemical libraries can access far greater chemical space, provided that the predictive accuracy is sufficient to identify useful molecules. Through the largest and most diverse virtual HTS campaign reported to date, comprising 318 individual projects, we demonstrate that our AtomNet® convolutional neural network successfully finds novel hits across every major therapeutic area and protein class. We address historical limitations of computational screening by demonstrating success for target proteins without known binders, high-quality X-ray crystal structures, or manual cherry-picking of compounds. We show that the molecules selected by the AtomNet® model are novel drug-like scaffolds rather than minor modifications to known bioactive compounds. Our empirical results suggest that computational methods can substantially replace HTS as the first step of small-molecule drug discovery