Journal: bioRxiv

Article Title: Network analysis reveals a distinct axis of macrophage activation in response to conflicting inflammatory cues

doi: 10.1101/844464

Figure Lengend Snippet: A) Dynamics of predicted gene expression in response to stimulation with LPS+IFNγ or IL4. Stimuli were added at 0 h. B) Kinetics of selected mRNAs in response to stimulation with LPS+IFNγ or IL4. C) mRNA expression profiles predicted by the model, validated against RNA-Seq measurements from peritoneal macrophages treated with LPS+IFNγ or IL4 for 4h. For semi-quantitative comparison between model and experiment, the log2 fold change of each mRNA vs. control was normalized by the root mean square between the M1 and M2 conditions. Classic M1 (orange) and M2 (green) phenotype markers are highlighted.

Article Snippet: Macrophages were assigned to one of three treatment groups: 1) stimulated with 1 μg/mL LPS (Sigma, L2880) and 20 ng/mL IFNγ (R&D, 485-MI) for 4 h; 2) stimulated with 20 ng/mL IL4 (R&D, 404-ML) for 4 h; or 3) untreated for 4 h, serving as the negative control.

Techniques: Gene Expression, Expressing, RNA Sequencing, Comparison, Control

Journal: bioRxiv

Article Title: Network analysis reveals a distinct axis of macrophage activation in response to conflicting inflammatory cues

doi: 10.1101/844464

Figure Lengend Snippet: A) Overall network influence of node knockdowns under stimulation with either LPS+IFNγ (orange) or IL4 (green). Nodes were ranked by the overall influence of their knockdown on all other network nodes, under conditions of LPS+IFNγ stimulation. B) Predicted effect of knockdown of influential nodes on activity of highly sensitive nodes, under conditions of LPS+IFNγ or IL4 treatment.

Article Snippet: Macrophages were assigned to one of three treatment groups: 1) stimulated with 1 μg/mL LPS (Sigma, L2880) and 20 ng/mL IFNγ (R&D, 485-MI) for 4 h; 2) stimulated with 20 ng/mL IL4 (R&D, 404-ML) for 4 h; or 3) untreated for 4 h, serving as the negative control.

Techniques: Knockdown, Activity Assay

Journal: bioRxiv

Article Title: Network analysis reveals a distinct axis of macrophage activation in response to conflicting inflammatory cues

doi: 10.1101/844464

Figure Lengend Snippet: A) Model-predicted signaling module activities in response to IFNγ and IL4 treatments at 4h, column normalized. B) Experimental validation of mRNA expression predicted in response to IFNγ, IL4, or IFNγ+IL4. For both experimental data and model predictions, mRNA were independently normalized by RMS-normalized log2 fold change at 4 h. C) Predicted expression dynamics of selected mRNAs in response to IFNγ, IL4, or IFNγ+IL4. D) Context-dependent network response to node knockdowns under treatments of IFNγ, IL4, or IFNγ+IL4.

Article Snippet: Macrophages were assigned to one of three treatment groups: 1) stimulated with 1 μg/mL LPS (Sigma, L2880) and 20 ng/mL IFNγ (R&D, 485-MI) for 4 h; 2) stimulated with 20 ng/mL IL4 (R&D, 404-ML) for 4 h; or 3) untreated for 4 h, serving as the negative control.

Techniques: Biomarker Discovery, Expressing

Journal: BJA: British Journal of Anaesthesia

Article Title: Maresin 1 attenuates neuroinflammation in a mouse model of perioperative neurocognitive disorders

doi: 10.1016/j.bja.2018.10.062

Figure Lengend Snippet: Anti-inflammatory and pro-resolving effects of maresin 1 prophylaxis. (a) Study design and endpoints. (b–e) Time course of plasma levels of interleukin (IL)-6, IL-12, CXCL1, and IL-10 at 24 h, 72 h, and 14 days after orthopaedic surgery. (f) Cognitive evaluation of mice using contextual fear conditioning. Training was performed before surgery and hippocampus-dependent memory was assessed at 72 h. MaR1 prevented surgery-induced cognitive dysfunction in both wild-type mice and (g) Ccr2 RFP/+ Cx3cr1 GFP/+ transgenic mice. (h–i) Effects of MaR1 on bone healing 14 days after stabilised tibia fracture surgery. Arrows indicate the original cortical bone. Scale bar: 50 μm. Data expressed as mean ( sem ); * P <0.05; n =5–6 per group (** P <0.01, n =8–10 for behaviour). C, Control; MaR1, Maresin 1; S, Surgery; sem , standard error of the mean.

Article Snippet: Separate batches of BMDMs were stimulated with LPS (10 ng ml −1 ) or 20 ng mL −1 recombinant mouse interleukin (IL)-4, IL-10, and recombinant human transforming growth factor (TGF)-β1 (R&D Systems) for 24 h and macrophage activation surface marker expression was assessed using flow cytometry.

Techniques: Transgenic Assay

Journal: BJA: British Journal of Anaesthesia

Article Title: Maresin 1 attenuates neuroinflammation in a mouse model of perioperative neurocognitive disorders

doi: 10.1016/j.bja.2018.10.062

Figure Lengend Snippet: Maresin 1 effects on bone marrow-derived macrophages (BMDMs). (a) Representative images of immunostaining of NF-κB in BMDMs. Stimulated with 10 ng ml −1 LPS for 2 h. (b) 2 h of LPS stimulation (10 ng ml −1 ) significantly increased NF-κB nuclear translocation in BMDMs. (c) LPS caused significant increase of TNF-α release from BMDMs after 24 h stimulation, which was reduced by co-application of MaR1 (10 nM). (d) NADPH oxidase mediated superoxide production in BMDMs was significantly increased after 24 h of LPS stimulation, which was inhibited by MaR1 co-application. (e–h) MaR1 reduced LPS-induced PD-L1 and CD86 expression in BMDMs at 24 h. No significant effect of MaR1 on PD-L2 and CD206 expression in BMDMs stimulated by M2 polarisation cytokines (IL-4/IL-10/TGF-β). Data are expressed as mean ( sem ). * P <0.05, ** P <0.01; n =3–5 per group. CLU, chemiluminescence unit; IL, interleukin; LPS, lipopolysaccharide; MaR1, Maresin 1; MFI, median fluorescence intensity; NADPH, nicotinamide adenine dinucleotide phosphate; sem , standard error of the mean; TNF, tumour necrosis factor.

Article Snippet: Separate batches of BMDMs were stimulated with LPS (10 ng ml −1 ) or 20 ng mL −1 recombinant mouse interleukin (IL)-4, IL-10, and recombinant human transforming growth factor (TGF)-β1 (R&D Systems) for 24 h and macrophage activation surface marker expression was assessed using flow cytometry.

Techniques: Derivative Assay, Immunostaining, Translocation Assay, Expressing, Fluorescence

Journal: BJA: British Journal of Anaesthesia

Article Title: Maresin 1 attenuates neuroinflammation in a mouse model of perioperative neurocognitive disorders

doi: 10.1016/j.bja.2018.10.062

Figure Lengend Snippet: Maresin 1 levels in CSF before, 24 h, and 6 weeks after major non-cardiac, non-neurologic surgery in older patients (age ≥60 yr). (a) Each line represents a single patient. Diagonal cross marks on the X and Y axes indicate non-linearity/scale discontinuity. n =11. (b) Working model for MaR1 protection in PNDs. Pre-treatment with MaR1 regulated excessive inflammation from circulating macrophage by reducing NF-κB activation, oxidative stress, pro-inflammatory cytokine release, overall contributing to a ‘M2-like’ switch. This systemic milieu did not impair fracture healing or cause signs of immunosuppression. In fact, these systemic effects prevented loss of cld-5 expression at the blood–brain barrier (BBB) and macrophage infiltration into the brain parenchyma. Overall, dampening the postoperative neuroinflammation leads to improved cognitive function. CSF, cerebrospinal fluid; IL, interleukin; MaR1, Maresin 1; NF-κB, nuclear factor-kappa B; PNDs, perioperative neurocognitive disorders; ROS, reactive oxygen species.

Article Snippet: Separate batches of BMDMs were stimulated with LPS (10 ng ml −1 ) or 20 ng mL −1 recombinant mouse interleukin (IL)-4, IL-10, and recombinant human transforming growth factor (TGF)-β1 (R&D Systems) for 24 h and macrophage activation surface marker expression was assessed using flow cytometry.

Techniques: Activation Assay, Expressing