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Additional file 1: Supplemental methods. (PDF)

Additional file 2: Figure S1. (PDF) In vivo anti-MM efficacy and in vitro functional status of activated NK cells. A) Activated (5 × 105) CFSE+ NK cells obtained from splenocytes of C57BL/KaLwRij or PBS (No Cell) were i.v. transferred into MM-bearing mice 3 weeks after 5TGM1 cell injection as in Fig. 1. Tumor growth was determined by FACS analysis of intracellular IgG2b + (tumor) cells among BM cells at 48 h after transfer. B, C) Purified NK cells were activated with IL-15, IL-12/15/18 or IL-15 (10 ng/ml: used as control) for 20 h and were incubated with or without 5TGM-1 cells (E:T ratio 1:1), and with anti-NKG2D or isotype control (i.c.) mAbs. B) NK cell degranulation was assessed by FACS analysis of % CD107a + cell. Left, representative dot plots showing the frequency of CD107a+ on NK cells. Right, average values ± SEM of CD107a+ cell frequency upon 5TGM-1 and anti-NKG2D mAb stimulation subtracted of degranulation in the absence of target cells or of i.c., respectively. Degranulation of control cells with tumor: 3%; degranulation of control cells with i.c. mAb: 5%). C) Upper panels: representative histogram plot showing NKG2D expression by activated NK cells (left) and average mean fluorescence intensity (MFI) values ± SEM (right); lower panel: cytotoxic activity of activated NK cells was measured by FACS analysis upon 6 h co-incubation with CFSE+ 5TGM1 cells and staining of dead cells with 7-AAD. D) Production of IFN-γ was assessed by FACS. Left panel, representative dot plots showing the frequency of IFN-γ+ NK cells. Right panel, average values ± SEM of IFN-γ+ cell frequency upon anti-NKG2D and i.c. mAbs stimulation. IFN-γ-producing control NK cells: 3%. Student t test was performed to compare differences of IFN-γ+ cell frequency between cells incubated with i.c. or anti-NKG2D mAb. Results in B, C and D are representative of three independent experiments.

Additional file 3: Figure S2. (PDF) CXCR4 expression by Cxcr3+/+ and Cxcr3−/− NK cells. Freshly purified, IL-15 and IL-12/15/18 activated (20 h) Cxcr3+/+ and Cxcr3−/− NK cells were stained for CXCR4 or isotype control. Upper panels show histogram plot of overlays of CXCR4 staining in untreated and cytokine treated cells of a representative analysis. White filled histograms represent isotype control (i.c.) staining. Lower panels show average ± SEM of median fluorescence intensity (MFI) from 3 independent analysis.

Additional file 4: Figure S3. (PDF) Anti-MM efficacy of IL-15 activated WT versus Cxcr3 deficient NK cells. A) Activated NK cells (5 × 105) from Cxcr3+/+ or Cxcr3−/− mice were transferred to mice two weeks after 5 T33 cell injection and tumor burden was calculated after 48 h. Graph shows the average ± SEM of frequency of tumor cells in BM and spleen from two independent experiments using a total of at least 4 animals per group. One-way ANOVA test was used to compare multiple groups. *, P

Additional file 5: Figure S4. (PDF). In vitro and in vivo expression kinetics of chemokine receptors on activated NK cells. A) Activated NK cells were labeled with 2.5 μM CFSE and adoptively transferred in mice 3 weeks after tumor cell injection following the experimental protocols depicted in Figs. 1 and 5. BM cells were isolated after 2 and 7 days and labeled with anti-CXCR4 mAb or isotype control along with anti-CD3 and anti-NK1.1. CXCR4 expression was evaluated on CFSE+ NK cells by FACS analysis. Left panels: representative histogram plots showing CXCR4 (Filled grey) expression by activated donor NK cells versus isotype control (filled white) staining. Right panels: average values ± SEM of MFI (n = 4 in two independent experiments). C) Purified NK cells were activated with IL-15, IL-12/15/18 for 20 h (control cells: IL-15 10 ng/ml), washed and rested in medium supplemented with IL-15 (10 ng/ml). Cells were harvested at the indicated time points and CXCR3 and CXCR4 expression was determined by FACS analysis. Results represent average values ± SEM of MFI of at least two independent analysis.

Additional file 6: Figure S5. (PDF). Chemokine expression in BM and tumor cells upon transfer of activated NK cells. Activated NK cells were labeled with 2.5 μM CFSE and adoptively transferred in mice 3 weeks after tumor cell injection following the experimental protocols depicted in Figs. 1 and 5. BM cells were isolated after 2 and 7 days. A) Left panels: NK cell number was determined by FACS analysis of CD3-NK1.1+ cells within donor CFSE- cells. Right panels: CXCL10 expression was determined in the bone marrow extracellular fluids from MM-bearing mice by ELISA. Histograms show mean values ± SEM of CXCL10 concentration (n = 4 from two experiments). B) Tumor cells were purified from pooled BM of tumor-bearing mice upon adoptive transfer of activated NK cells or PBS injection (no cell) and upon recombinant IL-15 administration (n = 3 mice group) at 2 days 7 days after adoptive transfer, respectively. CXCL10 and CXCL12 expression was measured by intracellular staining and FACS analysis (histogram plots) or by ELISA.

Converse magnetoelectric coupling in artificial multiferroics is generally modelled through three possible mechanisms: charge transfer, strain mediated or ion migration. Here we demonstrate a novel and highly reliable approach, where electrically controlled morphological modifications control the ferromagnetic response of a magnetoelectric heterostructure, specifically FexMn1-x ferromagnetic films on ferroelectric PMN-PT (001) substrates. The ferroelectric PMN-PT substrates present, in correspondence to electrical switching, fully reversible morphological changes at the surface, to which correspond reproducible modifications of the ferromagnetic response of the FexMn1-x films. Topographic analysis by atomic force microscopy shows the formation of surface cracks after application of a positive electric field up to 6 kV/cm, which disappear after application of negative voltage of the same amplitude. In-operando x-ray magnetic circular dichroic spectroscopy at Fe edge in FexMn1-x layers shows local variations of dichroic signal up to a factor 2.5 as a function of the electrically-driven morphological state. These findings highlight the role of morphology and surface topography as a key aspect in magnetoelectric coupling, whose proof of electrically reversible modification of the magnetic response adds a new possibility in the design of multiferroic heterostructures with electrically controlled functionalities.

Although, recently a mounting interest in the literature has been directed to sarcopenic obesity, currently, different definitions of sarcopenic obesity (SO) exist and diagnostic criteria are not universally established. The aim of the present systematic review is to summarize the state of the art according to extant literature about this topic. A systematic review of the literature was performed considering the revision question: “sarcopenic obesity: body composition”. Analysis and presentation of the outcomes were carried out in the form of a narrative resume of the reports. Twenty-six studies were considered eligible for the systematic review involving globally more than 23,000 subjects. In all the studies examined, except one, the definition of sarcopenic obesity was based on the co-presence of obesity and sarcopenia. With respect to obesity, different parameters (body mass index, fat mass and visceral fat area) were used (as single parameters or in association to the other adiposity measures). For sarcopenia, the appendicular skeletal muscle mass standardized for height or weight, cross-sectional area at the quadriceps level, muscle strength and/or muscle/fat-free mass indexes were considered. Body composition analysis was performed, depending on the studies, through DXA scan, CT scan bioimpedance analysis, dynamometer test or anthropometric parameters. To date, just few papers provided clear and shared diagnostic criteria of SO. In fact, in our review, criteria adopted by single authors were only partially overlapping. One of the main obstacles is represented by the definition of SO, and especially whether it should be based only on criteria of body composition or also functional criteria should be taken into account.

3 páginas, 2 figuras.-- et al., The reversal mechanisms of graded anisotropy FePtCu films have been investigated by alternating gradient magnetometer (AGM) and magneto-optical Kerr effect (MOKE) measurements with first-order reversal curve (FORC) techniques. The AGM-FORC analysis, which clearly shows the presence of soft and hard components, is unable to resolve how these phases are distributed throughout the film thickness. MOKE-FORC measurements, which preferentially probe the surface of the film, reveal that the soft components are indeed located toward the top surface. Combining AGM-FORC with the inherent surface sensitivity of MOKE-FORC analysis allows for a comprehensive analysis of heterogeneous systems such as graded materials., Support from The Swedish Foundation for strategic Research (SSF), The Swedish Research Council (VR), the Göran Gustafsson Foundation, and the Knut and Alice Wallenberg Foundation is gratefully acknowledged. V.B. thanks the Blanceflor Boncompagni–Ludovisi Foundation. J.N. thanks the Catalan DGR (Grant No. 2009SGR1292) and the Spanish MICINN (Grant No. MAT2010-20616-C02) projects for partial financial support. J.Å. is a Royal Swedish Academy of Sciences Research Fellow supported by a grant from the Knut and Alice Wallenberg Foundation.