Mean values for each session with standard deviations (SD), absolute and percentage smallest real difference (SRD) and intraclass correlation coefficient (ICC) for each motor performance parameter.

<p>SV = Spontaneous Velocity condition. MV = Maximal Velocity condition. 2 Hz = metronome condition (tone set at a rate of 2 Hz). 2 Hz_bim = metronome condition with both hands simultaneously. RATE = movement speed. TD = Touch Duration. ITI = Inter Tapping Interval. IHI = Inter Hand Interval. SD = Standard Deviation. SRD = Smallest Real Difference. ICC = Intraclass Correlation Coefficient.</p

ROC analysis.

<p>The ability of the model of discriminating patients with MS from the HC group is demonstrated by the area under the ROC curve (AUC = 0.89 (95%CI = 0.820.96, p<0.001)), even when evaluated using a cross-validation procedure based on the leave-one-out method (AUC = 0.88 (95%CI = 0.81–0.95, p<0.001)).</p

Comparison of the motor performance parameters between the two groups (HC and MS).

<p>(A) RATE in the spontaneous and maximal velocity conditions, (B) Touch Duration in the 2 Hz condition, (C) Inter Tapping Interval in the 2 Hz condition, (D) Inter Hand Interval in the 2 Hz_bim condition. * indicates statistically significant difference between the two groups (HC and MS). Error bars indicate standard error of the mean.</p

Additional file 1 of Microglia-derived CCL2 has a prime role in neocortex neuroinflammation

Additional file 1: Figure S1. Representative confocal microscopy images of immunolocalization of MSCs in the lung of EAE-affected MSC-treated mice (cs 1.5, 2.25). MSCs were retrovirally transduced to permanently express green fluorescent protein (GFP) and delivered intravenously to mice. At 24 h from disease onset/MSC treatment, a low GFP inherent fluorescence is detectable (a, arrows), whereas IHC with an anti-GFP antibody (b, c) shows distinct rounded cell profiles, with a large cytoplasm/nucleus ratio and a GFP-positive, fluorescent granular pattern (macrophage autofluorescence was below the photography threshold) in the lung septa. TOPRO-3 nuclear counterstaining. Scale bars: a, b 20 µm; c 10 µm

Correlations between the motor performance parameters and the clinical scores in the MS group, with age as control variable (correlation coefficient r values and p values are reported).

*<p>indicates statistical significance.</p><p>SV = Spontaneous Velocity condition. MV = Maximal Velocity condition. 2 Hz = metronome condition (tone set at a rate of 2 Hz). 2 Hz_bim = metronome condition with both hands simultaneously. RATE = movement speed. TD = Touch Duration. ITI = Inter Tapping Interval. IHI = Inter Hand Interval. EDSS = Expanded Disability Status Scale. MSFC = Multiple Sclerosis Functional Composite. 9-HPT = 9-Hole Peg Test. T25W = Timed 25-Foot Walk. PASAT = Paced Auditory Serial Addition. MFIS = Modified Fatigue Impact Scale.</p

Test-retest reliability of the motor performance parameters.

<p><b>(A-E)</b> Motor performance parameters in the two sessions performed one-month apart in a group of healthy controls (trial 2 vs. trial 1), with respect to the identity (i.e., the dashed line represents the bisector).</p

Expression of CXCR3 and interferon (IFN)-γ by (SF) CCR7and CCR7memory CD4cells from synovial fluid

<p><b>Copyright information:</b></p><p>Taken from "Phenotypic and functional characterisation of CCR7and CCR7CD4memory T cells homing to the joints in juvenile idiopathic arthritis"</p><p>Arthritis Research & Therapy 2005;7(2):R256-R267.</p><p>Published online 12 Jan 2005</p><p>PMCID:PMC1065323.</p><p>Copyright © 2005 Gattorno et al.; licensee BioMed Central Ltd.</p> IFN-γ expression was investigated by three-colour staining of freshly isolated SF CD45ROcells with CD4–fluorescein isothiocyanate (FITC), anti-CCR7–phycoerythrin (PE) and anti-CCR5–CyChrome monoclonal antibodies (mAbs) or CD4–TC (where TC stands for Tri-color), anti-CCR7–PE and anti-IFN-γ mAbs, respectively; CXCR3 expression was investigated by triple staining with CD4–TC, anti-CCR7–PE and anti-CXCR3–FITC, as described in the Methods section. Subsequently, cytofluorimetric analysis was performed by gating on the CD4CCR7and CD4CCR7lymphocyte subsets. Data are expressed as percentages of positive cells or/and mean fluorescence intensity. Expression of IFN-γ (a) and CXCR3 (b) by SF CCR7and CCR7memory CD4cells from 10 patients with juvenile idiopathic arthritis (JIA). Boxes contain values falling between the 25th and 75th centiles; whiskers show lines that extend from the boxes represent the highest and lowest values for each subgroup. Differences between paired SF mononuclear cells were evaluated by the Wilcoxon rank test. Dot plots show the cytofluorimetric analysis IFN-γ (c) and CXCR3 (d) expression by the gated CD4CCR7(gate 1) and CD4CCR7(gate 2) cell populations in three representative patients with JIA

DataSheet_1_Extracellular vesicles released by microglia and macrophages carry endocannabinoids which foster oligodendrocyte differentiation.pdf

IntroductionMicroglia and macrophages can influence the evolution of myelin lesions through the production of extracellular vesicles (EVs). While microglial EVs promote in vitro differentiation of oligodendrocyte precursor cells (OPCs), whether EVs derived from macrophages aid or limit OPC maturation is unknown.MethodsImmunofluorescence analysis for the myelin protein MBP was employed to evaluate the impact of EVs from primary rat macrophages on cultured OPC differentiation. Raman spectroscopy and liquid chromatography-mass spectrometry was used to define the promyelinating lipid components of myelin EVs obtained in vitro and isolated from human plasma.Results and discussionHere we show that macrophage-derived EVs do not promote OPC differentiation, and those released from macrophages polarized towards an inflammatory state inhibit OPC maturation. However, their lipid cargo promotes OPC maturation in a similar manner to microglial EVs. We identify the promyelinating endocannabinoids anandamide and 2-arachidonoylglycerol in EVs released by both macrophages and microglia in vitro and circulating in human plasma. Analysis of OPC differentiation in the presence of the endocannabinoid receptor antagonists SR141716A and AM630 reveals a key role of vesicular endocannabinoids in OPC maturation. From this study, EV-associated endocannabinoids emerge as important mediators in microglia/macrophage-oligodendrocyte crosstalk, which may be exploited to enhance myelin repair.</p

Activated T cells undergo massive NAD⁺ depletion upon Nampt inhibition.

<p>A, 3×10⁶ PBLs/well were stimulated (or not, unstim.) with 5 µg/ml PHA, 1 µg/ml con A, or 50 ng/ml PMA and 0.5 µM ionomycin in the presence or absence of the indicated FK866 concentrations. 48 h later, cells were lysed in 0.6 M PCA and NAD⁺ content was measured in neutralized extracts. NAD⁺ levels were normalized to those detected in the absence of FK866. B, Unstimulated or PHA-stimulated PBLs were treated with 33 nM FK866 for 48 h before NAD⁺ content was determined. Absolute NAD⁺ levels are presented. *: p<0.05. C, PBLs were cultured for 48 h with PHA with or without FK866 (33 nM) addition. Subsequently, pyridine dinucleotides levels were measured in acid (NAD⁺ and NAPD⁺) or alkaline (NADH and NADPH) cell extracts. Dinucleotide levels were normalized to those detected without FK866. D, PBLs were incubated with PHA and 33 nM FK866 for the indicated times. Thereafter cells were harvested and NAD⁺ and ATP levels were determined in cell extracts whereas cell viability was assessed by PI-staining and flow cytometry. Results were normalized to the values of FK866-untreated cells. E, Resting or PHA-stimulated PBLs were treated (or not) with 33 nM FK866 in the presence or absence of 1 mM NAD⁺. After five-days viability was assessed determining PI⁻ cells by flow cytometry. Results are means ± SD of five (A) or three (B–E) experiments.</p

PARP inhibitors and sirtinol attenuate FK866-induced T cell demise.

<p>A, PBLs were cultured for 24 h with or without PHA. Thereafter, Nampt and PARP1 levels were detected by Q-PCR. mRNA levels in PHA-stimulated cells were compared to those in unstimulated PBLs. B, Resting or PHA-stimulated PBLs were incubated with or without 33 nM FK866 in the presence or absence of 300 µM NU1025, 10 µM PJ34, or 300 µM 3-AB. 48 h later NAD⁺ levels were assessed (presented as % of values in FK866-untreated PBLs). *, p<0.05. C, PHA-stimulated PBLs were incubated for five days with or without 33 nM FK866 in the presence or absence of 300 µM NU1025, 10 µM PJ34, or 300 µM 3-AB. Thereafter, viability was assessed. D, 5×10⁵ Jurkat cells were treated for two days with 500 pM FK866 in the presence or absence of 300 µM NU1025, 5 µM PJ34, or 300 µM 3-AB. Subsequently, NAD⁺ content was determined and expressed as % of values in FK866-untreated Jurkat. E, 3×10⁴ Jurkat cells/well were incubated in 96-well plates with or without 300 pM FK866 in the presence or absence of the indicated concentrations of NU1025, PJ34, or 3-AB. Viability was determined 96 h later by PI cell staining and flow cytometry. F, PBLs were incubated for five days with PHA, with or without 33 nM FK866, in the presence or absence of 30 µM sirtinol. Viability was subsequently assessed by PI staining and flow cytometry. C, E, F, each treatment was tested in triplicate wells. Results are presented as means ± SD of three experiments.</p