Signaling in natural killer cells: SHIP, 2B4 and the Kinome

The NK cell is a large granular lymphocyte that plays a key role in protecting the body against numerous pathogens including parasites, intracellular bacteria, viral infections, as well as showing anti-tumor activity and playing a role in the rejection of allogeneic BM. Unlike other lymphocytic cell types, that utilize rearranging receptors, NK cells are regulated by a complex array of germ line encoded activating and inhibitory receptors. NK cells are often described as a front line or rapid defense given their response to stimuli can be immediate, although they also maintain functions that extend their role well into the adaptive immune system. Inhibitory receptors that recognize MHC class I molecules regulate NK cell responses and self-tolerance. Recent evidence indicates self-ligands not present in the MHC locus can also modulate NK function. We previously demonstrated that the NK receptor repertoire is disrupted by SHIP-deficiency. Here we show that an inhibitory receptor, 2B4, that recognizes an MHC-independent ligand is over expressed in NK cells of SHIP-/- mice at all stages of NK development and differentiation. Overexpression of 2B4 compromises key cytolytic NK functions, including killing of allogeneic, tumor and viral targets. These results demonstrate that in SHIP-/- NK cell 2B4 is the dominant inhibitory receptor. We then furthered this finding by examining the molecular basis of 2B4 dominance. We show that in SHIP-/- NK cells there is increased 2B4 expression as well as a strong bias towards the 2B4L isoform. We have also identified a greater than tenfold increase in SHP1 recruitment to 2B4. Consistent with this SHP1 over recruitment,both a broad and a selective SHP1 inhibitor restore SHIP-/- NK killing of complex targets.Through this study we have identified the molecular mechanism of 2B4 receptor dominance as SHP1 over-recruitment.In addition we have utilized protein array technology to explore NK signaling through the determination of the NK kinome. To this end we have been able to identify multiple pathways that may mark crucial differences between the mature and immature NK cell

1H NMR Spectroscopy and MVA Analysis of Diplodus sargus Eating the Exotic Pest Caulerpa cylindracea

6noThe green alga Caulerpa cylindracea is a non-autochthonous and invasive species that is severely affecting the native communities in the Mediterranean Sea. Recent researches show that the native edible fish Diplodus sargus actively feeds on this alga and cellular and physiological alterations have been related to the novel alimentary habits. The complex effects of such a trophic exposure to the invasive pest are still poorly understood. Here we report on the metabolic profiles of plasma from D. sargus individuals exposed to C. cylindracea along the southern Italian coast, using 1H NMR spectroscopy and multivariate analysis (Principal Component Analysis, PCA, Orthogonal Partial Least Square, PLS, and Orthogonal Partial Least Square Discriminant Analysis, OPLS-DA). Fish were sampled in two seasonal periods from three different locations, each characterized by a different degree of algal abundance. The levels of the algal bisindole alkaloid caulerpin, which is accumulated in the fish tissues, was used as an indicator of the trophic exposure to the seaweed and related to the plasma metabolic profiles. The profiles appeared clearly influenced by the sampling period beside the content of caulerpin, while the analyses also supported a moderate alteration of lipid and choline metabolism related to the Caulerpa-based diet.openopenS. A. De Pascali; L. Del Coco; S. Felline; E. Mollo; A. Terlizzi; F. P. FanizziS. A., De Pascali; L., Del Coco; S., Felline; E., Mollo; Terlizzi, Antonio; F. P., Fanizz

Harvest year effects on Apulian EVOOs evaluated by (1)H NMR based metabolomics

none5noNine hundred extra virgin olive oils (EVOO) were extracted from individual olive trees of four olive cultivars (Coratina, Cima di Mola, Ogliarola, Peranzana), originating from the provinces of Bari and Foggia (Apulia region, Southern Italy) and collected during two consecutive harvesting seasons (2013/14 and 2014/15). Following genetic identification of individual olive trees, a detailed Apulian EVOO NMR database was built using 900 oils samples obtained from 900 cultivar certified single trees. A study on the olive oil lipid profile was carried out by statistical multivariate analysis (Principal Component Analysis, PCA, Partial Least-Squares Discriminant Analysis, PLS-DA, Orthogonal Partial Least-Squares Discriminant Analysis, OPLS-DA). Influence of cultivar and weather conditions, such as the summer rainfall, on the oil metabolic profile have been evaluated. Mahalanobis distances and J2 criterion have been measured to assess the quality of resulting scores clusters for each cultivar in the two harvesting campaigns. The four studied cultivars showed non homogeneous behavior. Notwithstanding the geographical spread and the wide number of samples, Coratina showed a consistent behavior of its metabolic profile in the two considered harvests. Among the other three Peranzana showed the second more consistent behavior, while Cima di Mola and Ogliarola having the biggest change over the two years.Girelli, Chiara R; Del Coco, Laura; Papadia, Paride; De Pascali, Sandra A; Fanizzi, Francesco PGirelli, CHIARA ROBERTA; DEL COCO, Laura; Papadia, Paride; DE PASCALI, SANDRA ANGELICA; Fanizzi, Francesco Paol

Enhanced Bioactivity of Pomegranate Peel Extract following Controlled Release from CaCO3 Nanocrystals

none9Pomegranate peel extract is rich of interesting bioactive chemicals, principally phenolic compounds, which have shown antimicrobial, anticancer, and antioxidative properties. The aim of this work was to improve extract’ bioactivity through the adsorption on calcium carbonate nanocrystals. Nanocrystals revealed as efficient tools for extract adsorption reaching 50% of loading efficiency. Controlled release of the contained metabolites under acidic pH has been found, as it was confirmed by quantitative assay and qualitative study through NMR analysis. Specific functionality of inorganic nanocarriers could be also tuned by biopolymeric coating. The resulting coated nanoformulations showed a great antimicrobial activity against B. cinerea fungus preventing strawberries disease better than a commercial fungicide. Furthermore, nanoformulations demonstrated a good antiproliferative activity in neuroblastoma and breast cancer cells carrying out a higher cytotoxic effect respect to free extract, confirming a crucial role of nanocarriers. Finally, pomegranate peel extract showed a very high radical scavenging ability, equal to ascorbic acid. Antioxidant activity, measured also in intracellular environment, highlighted a protective action of extract-adsorbed nanocrystals twice than free extract, providing a possible application for new nutraceutical formulations.Regione Puglia, Project Research for Innovation (REFIN) “Sintesi di un sistema teranostico a base di nano-cellulosa per la detection e la cura dei tumori” Ministero dello Sviluppo Economico, bando “AGRIFOOD” D.M. 5 marzo 2018 Capo III. Prog. N. F/200060/01-03/X45 PERSEFONE - Punica granatum e Nanotecnologie: una value chain per la valorizzazione degli scarti e sottoprodotti finalizzata alla produzione di integratori alimentari e compost. PON Ricerca e Innovazione 2014-2020 - Avviso per la presentazione di progetti dei Ricerca Industriale e Sviluppo Sperimentale nelle 12 aree di specializzazione individuate dal PNR 2015 – 2020, di cui al Decreto Direttoriale MIUR del 13 luglio 2017, n. 1735 NanotEcnologie chiMiche green per la protEzione Sostenibile delle pIante (NEMESI) ARS01_01002 Area di Specializzazione “Chimica Verde” CUP: F36C18000180005, Ministero dell'Università e Ricerca. “Olivicoltura e difesa da Xylella fastidiosa e da insetti vettori in Italia - (Oli.Di.X.I.It)”, prot. Mipaaf n.0011485 del 05/04/2017 Ministry of Agricultural, Food and Forestry Policies.openFrancesca Baldassarre; Viviana Vergaro; Federica De Castro; Francesca Biondo; Gian Paolo Suranna; Paride Papadia; Francesco P. Fanizzi; Domenico Rongai; Giuseppe CiccarellaBaldassarre, Francesca; Vergaro, Viviana; DE CASTRO, Federica; Biondo, Francesca; Suranna, Gian Paolo; Papadia, Paride; Fanizzi, Francesco P.; Rongai, Domenico; Ciccarella, Giusepp

Square-Planar PtII versus Octahedral PtIV Halido Complexes:195Pt NMR Explained by a Simple Empirical Approach

In this work, we show by a simple empirical approach that a linear relationship between observed 195Pt NMR frequencies and the overall sum of the ionic radii of the coordinated halido ligands [Σ(rh)] exists in square-planar PtII complexes of the type [PtXnY4–n]2– (1 ≤ n≤4; X, Y = Cl, Br, I). Another finding was that such square-planar complexes could be empirically described as octahedral complexes, with the two lobes of the 5dz2 orbital above and below the coordination plane acting as two pseudo-halido ligands, each showing a constant apparent radius of around 207 pm. According to our approach, the overall apparent radius of around 415 pm produces constant 195Pt NMR shielding for all [PtXnY4–n]2– complexes of about 10450 ppm. This result is 1) consistent with the theoretically calculated overall 5d shell lone-pair shielding observed in square-planar PtII with respect to octahedral PtIV complexes and 2) almost coincident with the already measured chemical shift anisotropy (CSA) of the K2[PtCl4] complex both in solution and in the solid state (single crystal)

H-1 Nuclear Magnetic Resonance Study of Olive Oils Commercially Available as Italian Products in the United States of America

Multivariate analysis of H-1 NMR data has been used for the characterization of 12 blended olive oils commercially available in the U. S. as Italian products. Chemometric methods such as unsupervised Principal Component Analysis (PCA) allowed good discrimination and gave some affinity indications for the U. S. market olive oils compared to other single cultivars of extra virgin olive oil such as Coratina and Ogliarola from Apulia, one of Italy's leading olive oil producers, Picual (Spain), Kalamata (Greece) and Sfax (Tunisia). The olive oils commercially available as Italian products in the U. S. market clustered into 3 groups. Among them only the first (7 samples) and the second group (2 samples) showed PCA ranges similar to European references. Two oils of the third group (3 samples) were more similar to Tunisian references. In conclusion, our study revealed that most EVOO (extra virgin olive oils) tested were closer to Greek (in particular) and Spanish olive oils than Apulia EVOO. The PCA loadings disclose the components responsible for the discrimination as unsaturated (oleic, linoleic, linolenic) and saturated fatty acids. All are of great importance because of their nutritional value and differential effects on the oxidative stability of oils. It is evident that this approach has the potential to reveal the origin of EVOO, although the results support the need for a larger database, including EVOO from other Italian regions