Changing tools to catch the beast: Why the EU studies should take policy seriously, and how this shift could help to understand integration

While the EU is still enlarging its membership and range of actions, the current stalemate of the integration project is pushing the ‘ontological’ question about the nature of the common Europe again at the top of both the political and the research agendas. This paper aims to contribute the debate and display the possibilities of enhancing the comprehension of the ‘supranational beast’ from a policy perspective. The focus hence is shifted on implementation and policy frameworks, and the field of analysis widened to cover the institutional transformations occurred within the administrative dimension both at the national and supranational levels in the last decades. From this perspective, previous findings are revisited to account for the new meaning of the common Europe after the Single European Act, the complexity of the current institutional architecture, and the reasons beneath the stalemate. Finally, the approach is translated into research hypotheses about integration and viable strategies for sustaining it beneath and beyond the usual ‘hard’ institutional re-engineering

AIRE polymorphism, melanoma antigen-specific T cell immunity, and susceptibility to melanoma

AIRE is involved in susceptibility to melanoma perhaps regulating T cell immunity against melanoma antigens (MA). To address this issue, AIRE and MAGEB2 expressions were measured by real time PCR in medullary thymic epithelial cells (mTECs) from two strains of C57BL/6 mice bearing either T or C allelic variant of the rs1800522 AIRE SNP. Moreover, the extent of apoptosis induced by mTECs in MAGEB2-specific T cells and the susceptibility to in vivo melanoma B16F10 cell challenge were compared in the two mouse strains. The C allelic variant, protective in humans against melanoma, induced lower AIRE and MAGEB2 expression in C57BL/6 mouse mTECs than the T allele. Moreover, mTECs expressing the C allelic variant induced lower extent of apoptosis in MAGEB2-specific syngeneic T cells than mTECs bearing the T allelic variant (p < 0.05). Vaccination against MAGEB2 induced higher frequency of MAGEB2-specific CTL and exerted higher protective effect against melanoma development in mice bearing the CC AIRE genotype than in those bearing the TT one (p < 0.05). These findings show that allelic variants of one AIRE SNP may differentially shape the MA-specific T cell repertoire potentially influencing susceptibility to melanoma

Le regioni tra Stato ed Unione europea: l'emergere di un terzo livello?: il negoziato italiano sulle politiche di coesione 2000-2006: il caso delle regioni ob.2

Dottorato di ricerca in sociologia. 12. ciclo. Relatori A. Cavalli e G. E. RusconiConsiglio Nazionale delle Ricerche - Biblioteca Centrale - P.le Aldo Moro, 7, Rome; Biblioteca Nazionale Centrale - P.za Cavalleggeri, 1, Florence / CNR - Consiglio Nazionale delle RichercheSIGLEITItal

Rules as data

Rules lie at the core of many disciplines beneath regulatory studies. Such a broad interest inevitably comes with fragmented understandings and technical choices that hinder knowledge cumulation and learning. This introduction tackles these limitations through an encompassing analytical blueprint from measurement theory. First, it addresses ambiguities to establish formal rules as a distinct research object. Then, it builds on legal, institutional analytic, and computational linguistic frameworks to pinpoint their constituting elements. Last, it revises strategies for assigning meaningful numbers to objects and outlines how the contributions to this Special Issue foster different aspects of the blueprint

Synthesis, structural characterization and effect on human granulocyte intracellular cAMP levels of abscisic acid analogs

The phytohormone abscisic acid (ABA), in addition to regulating physiological functions in plants, is alsoproduced and released by several mammalian cell types, including human granulocytes, where it stim-ulates innate immune functions via an increase of the intracellular cAMP concentration ([cAMP]i).We synthesized several ABA analogs and evaluated the structure\u2013activity relationship, by the system-atical modification of selected regions of these analogs. The resulting molecules were tested for their abil-ity to inhibit the ABA-induced increase of [cAMP]i in human granulocytes. The analogs with modifiedconfigurations at C-20and C-30abrogated the ABA-induced increase of the [cAMP]i and also inhibited sev-eral pro-inflammatory effects induced by exogenous ABA on granulocytes and monocytes. Accordingly,these analogs could be suitable as novel putative anti-inflammatory compounds. \ua9 2014 Elsevier Ltd. All rights reserved

Trastuzumab quantification in serum: a new, rapid, robust ELISA assay based on a mimetic peptide that specifically recognizes trastuzumab

Trastuzumab, a humanized monoclonal antibody directed against the epidermal growth factor receptor 2 (HER2), is a milestone in the treatment of HER2-overexpressing breast cancer patients. An enzyme-linked immunosorbent assay (ELISA) for trastuzumab has been developed for routine use in the laboratory to support clinical and pharmacokinetic studies to optimize therapy. The method relies on an antigen peptide linked to a 96-well plate via the streptavidin/biotin system. The peptide sequence mimics the extracellular portion of the HER2 receptor that is recognized by trastuzumab. The calibration range of the assay is 10 to 360 ng/mL per well, corresponding to a trastuzumab serum concentration from 5 to 180 mu g/mL with a lower limit of quantification of 10 mu g/mL. Validation results demonstrate that trastuzumab can be accurately and precisely quantified in human serum using this assay. The procedure was also tested in sera obtained from breast cancer patients to evaluate trastuzumab serum levels, confirming the applicability of method that could be a valid assay to use in daily laboratory practice

SIRT6 inhibitors with salicylate-like structure show immunosuppressive and chemosensitizing effects

The NAD+-dependent deacetylase SIRT6 is an emerging cancer drug target, whose inhibition sensitizes cancer cells to chemo-radiotherapy and has pro-differentiating effects. Here we report on the identification of novel SIRT6 inhibitors with a salicylate-based structure. The new SIRT6 inhibitors show improved potency and specificity compared to the hit inhibitor identified in an in silico compound screen. As predicted based on SIRT6 biological roles, the new leads increase histone 3 lysine 9 acetylation and glucose uptake in cultured cells, while blocking TNF-\uce\ub1 production and T lymphocyte proliferation. Notably, the new SIRT6 inhibitors effectively sensitize pancreatic cancer cells to gemcitabine. Finally, studies of compound fingerprinting and pharmacokinetics defined the drug-like properties of one of the new SIRT6 inhibitors, potentially allowing for subsequent in vivo proof-of-concept studies. In conclusion, new SIRT6 inhibitors with a salicylate-like structure were identified, which are active in cells and could potentially find applications in disease conditions, including cancer and immune-mediated disorders

SIRT6 deacetylase activity regulates NAMPT activity and NAD(P)(H) pools in cancer cells

Nicotinamide phosphoribosyltransferase (NAMPT) is the rate-limiting enzyme in the NAD+ salvage pathway from nicotinamide. By controlling the biosynthesis of NAD+, NAMPT regulates the activity of NAD+-converting enzymes, such as CD38, poly-ADP-ribose polymerases, and sirtuins (SIRTs). SIRT6 is involved in the regulation of a wide number of metabolic processes. In this study, we investigated the ability of SIRT6 to regulate intracellular NAMPT activity and NAD(P)(H) levels. BxPC-3 cells and MCF-7 cells were engineered to overexpress a catalytically active or a catalytically inactive SIRT6 form or were engineered to silence endogenous SIRT6 expression. In SIRT6-overexpressing cells, NAD(H) levels were up-regulated, as a consequence of NAMPT activation. By immunopurification and incubation with recombinant SIRT6, NAMPT was found to be a direct substrate of SIRT6 deacetylation, with a mechanism that up-regulates NAMPT enzymatic activity. Extracellular NAMPT release was enhanced in SIRT6-silenced cells. Also glucose-6-phosphate dehydrogenase activity and NADPH levels were increased in SIRT6-overexpressing cells. Accordingly, increased SIRT6 levels reduced cancer cell susceptibility to H2O2-induced oxidative stress and to doxorubicin. Our data demonstrate that SIRT6 affects intracellular NAMPT activity, boosts NAD(P)(H) levels, and protects against oxidative stress. The use of SIRT6 inhibitors, together with agents inducing oxidative stress, may represent a promising treatment strategy in cancer