Hallazgos numismáticos de época romana en Fiñana (Almería)

Presentamos un conjunto de 16 monedas aparecidas en Fiñana (Almería), dentro de los trabajos de recopilación de materiales numismáticos, que venimos efectuando desde el Grupo de Investigación dirigido por el Dr. González Román, con vistas a realizar ulteriores estudios sobre circulación monetaria y comercio en el sureste peninsular.The autors of this paper present thirty three roman coins from Huétor Tájar, a village in the western part of Granada's fertile plain. The coins have a cronology among in the mid III century and at the end of the IV century a.Ch

Cardiac Extracellular Matrix Hydrogel Enriched with Polyethylene Glycol Presents Improved Gelation Time and Increased On-Target Site Retention of Extracellular Vesicles

Stem-cell-derived extracellular vesicles (EVs) have demonstrated multiple beneficial effects in preclinical models of cardiac diseases. However, poor retention at the target site may limit their therapeutic efficacy. Cardiac extracellular matrix hydrogels (cECMH) seem promising as drug-delivery materials and could improve the retention of EVs, but may be limited by their long gelation time and soft mechanical properties. Our objective was to develop and characterize an optimized product combining cECMH, polyethylene glycol (PEG), and EVs (EVs–PEG–cECMH) in an attempt to overcome their individual limitations: long gelation time of the cECMH and poor retention of the EVs. The new combined product presented improved physicochemical properties (60% reduction in half gelation time, p < 0.001, and threefold increase in storage modulus, p < 0.01, vs. cECMH alone), while preserving injectability and biodegradability. It also maintained in vitro bioactivity of its individual components (55% reduction in cellular senescence vs. serum-free medium, p < 0.001, similar to EVs and cECMH alone) and increased on-site retention in vivo (fourfold increase vs. EVs alone, p < 0.05). In conclusion, the combination of EVs–PEG–cECMH is a potential multipronged product with improved gelation time and mechanical properties, increased on-site retention, and maintained bioactivity that, all together, may translate into boosted therapeutic efficacy

Role of HLA-G in shaping Natural Killer cell biology in cancer

[eng] The human leukocyte antigen-G (HLA-G) is a non-classic MHC class I molecule. It is called “non-classic” because it differs from classic MHC class I molecules due to its very low amount of polymorphism and highly tissue-restricted expression (Figure 1). HLA-G have a role in this tolerogenic immune response that takes place at the maternal-fetal interface, by inhibiting cytotoxicity and cytokine production by NK and T cells. Moreover, apart from its physiological expression, HLA-G can also be expressed during pathological conditions such as cancer, where it also downregulates the immune response and allows the tumor progression. The natural killer (NK) cells are innate immune effectors that can spontaneously kill tumor cells through the secretion of cytolytic molecules such as granzymes and perforins. Moreover, they can be subdivided due to the cell surface density of CD56 in CD56bright and CD56dim. The CD56dim subset is the most abundant in peripheral blood (approximately 90%) and expresses high levels of CD16 whereas the minority of NKs are CD56bright CD16dim/negative. The CD56bright subset is described to have a regulatory role, whereas the CD56 dim is more cytotoxic (Figure 2). The aim of this work was to characterize the role of HLA-G in the regulation of the IS focusing in the NK cells and to study the biological implications in the context of cancer. Moreover, we explored the possibility of targeting the HLA-G molecule. We designed and performed an in vitro co-culture (CC) system to study the consequences of the presence or absence of HLA-G in the target cells (JEG3) in the immune effector cells (NK92). We observed an increase of the CD56 marker when the NK92 cells were co-cultured in the presence of HLA-G. We also developed another in vitro culture model to check the effect of the soluble-HLA-G in the NK92 cells. With this aim, we generated conditioned media (CM) from JEG3 and HuP-T3 cells and characterized the level of secreted-HLA-G present by ELISA. We used this CM as a physiological source of HLA-G and cultured the NK cells with it. We observed as well as in the CC experiments an increase of the CD56 marker in presence of the HLA-G (NK92+ CM form JEG3 WT). Importantly, we also observed the down-regulation of IFN-γ, granzyme and perforin secretion. We confirmed these results using recombinant-HLA-G protein. Also, the granzyme B and perforin secretion was reverted, however, we did not observe a reversion of the IFNγ secretion pattern. Furthermore, we tried to block the HLA-G effect by the addition of a monoclonal anti-HLA-G antibody (Thermofisher, clone 87G). However, due to the lack of effect that we obtained with this antibody and given cross-reactivity previously described in bibliography, we decided to design our own anti-HLA-G blocking antibody. We targeted a region in the alpha-3 domain described to be responsible for the ILT-2 receptor binding in order to functionally block the effect of HLA-G. The antibody we obtained, worked in a dose-dependent manner in NK92 reverting the effects produced by the soluble HLA-G. However, we did not have the same results with the pb-NK cells. In this work we have shown the implication of HLA-G in the regulation of the NK cell maturation and function. This is very important given the role of the NK cells in the context of the immune-modulation in cancer. Particularly, we have described for the first time, the role of HLA-G in the modulation of the CD56 surface marker. Interestingly, HLA-G induces a NK cell phenotype that recapitulates the one present at the decidua in the pregnancy context. This parallelism between cancer and pregnancy demonstrate us that once again cancer hijacks a preexisting mechanism for its own benefit. Moreover, we have explored the possibility of reverting this effect produced in NK cells using an anti-HLA-G antibody. However we described only partial effects, highlighting the fact that further work is required to evaluate HLA-G as a potential therapeutic target

Rapamycin negatively impacts insulin signaling, glucose uptake and uncoupling protein-1 in brown adipocytes

New onset diabetes after transplantation (NODAT) is a metabolic disorder that affects 40% of patients on immunosuppressive agent (IA) treatment, such as rapamycin (also known as sirolimus). IAs negatively modulate insulin action in peripheral tissues including skeletal muscle, liver and white fat. However, the effects of IAs on insulin sensitivity and thermogenesis in brown adipose tissue (BAT) have not been investigated. We have analyzed the impact of rapamycin on insulin signaling, thermogenic gene-expression and mitochondrial respiration in BAT. Treatment of brown adipocytes with rapamycin for 16 h significantly decreased insulin receptor substrate 1 (IRS1) protein expression and insulin-mediated protein kinase B (Akt) phosphorylation. Consequently, both insulin-induced glucose transporter 4 (GLUT4) translocation to the plasma membrane and glucose uptake were decreased. Early activation of the N-terminal Janus activated kinase (JNK) was also observed, thereby increasing IRS1 Ser 307 phosphorylation. These effects of rapamycin on insulin signaling in brown adipocytes were partly prevented by a JNK inhibitor. In vivo treatment of rats with rapamycin for three weeks abolished insulin-mediated Akt phosphorylation in BAT. Rapamycin also inhibited norepinephrine (NE)-induced lipolysis, the expression of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) and uncoupling protein (UCP)-1 in brown adipocytes. Importantly, basal mitochondrial respiration, proton leak and maximal respiratory capacity were significantly decreased in brown adipocytes treated with rapamycin. In conclusion, we demonstrate, for the first time the important role of brown adipocytes as target cells of rapamycin, suggesting that insulin resistance in BAT might play a major role in NODAT development.This work was supported by Ministerio de Economía y Competitividad: (SAF2015-65267-R to AMV, SAF2012-32491 to MJO, SAF2013-45887-R to LH, and SAF2014-52223-C2-1-R to DS) that were cofunded by the Fondos Europeos de Desarrollo Regional de la Unión Europea [FEDER], Spain, Instituto de Salud Carlos III (FIS PI15-00448 to SC and INFLAMES PIE14/00045 to AMV), Centro de Investigación Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas (CIBERdem, ISCIII, Spain) to AMV, Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y la Nutrición (CIBERobn-Grant CB06/03/0001) to DS and S2010/BMD-2423 (Comunidad de Madrid, Spain) to AMV and MJO and S2010/BMD-2402 to SC and 2014SGR465 (Generalitat de Catalunya, Spain) to DS. CdM was supported by the Albert Reynold Grant travel fellowship from the European Foundation for the Study of Diabetes. EC and FR were funded by FEDER through the Operational Programme Competitiveness Factors - COMPETE and by FCT - Foundation for Science and TechnologyPTDC/SAU-OSM/104124/2008 and strategic project UID/NEU/04539/2013.Peer Reviewe

Sirolimus negatively impacts insulin signaling, glucose uptake and uncoupling protein-1 in brown adipocytes

Resumen del póster presentado presentado al CIBERDEM Annual Meeting, celebrado en Cerdanyola del Vallès, Barcelona (España) del 11 al 13 de mayo de 2016.-- et al.New onset diabetes after transplantation (NODAT) is a metabolic syndrome that affects a large number of patients with chronic treatment with immunosuppressive agents (IAs) such as sirolimus (also known as rapamycin). IAs negatively modulate insulin sensitive-tissues such as skeletal muscle, liver and white fat. However, the effects of IAs on insulin action and thermogenesis in brown adipose tissue (BAT) have not yet been investigated. In this study, we have analyzed the impact of sirolimus in the insulin signaling and thermogenic gene-expression in brown adipocytes (BA). We found that sirolimus induced the degradation of insulin receptor substrate 1 (IRS1) leading to a decrease in insulin-mediated protein kinase B (Akt) phosphorylation. As a consequence, in sirolimus-treated BA, insulin-induced glucose transporter 4 (GLUT4) translocation to the plasma membrane and glucose uptake were decreased. Sirolimus triggered an early activation of N-terminal Janus activated kinase (JNK) thereby increasing serine 307 phosphorylation of IRS1 that preceded its proteasomal degradation. The negative effects of sirolimus on insulin signaling were prevented by a JNK inhibitor. In vivo treatment of rats with sirolimus for three weeks abolished insulin-mediated Akt phosphorylation in BAT. Sirolimus also inhibited norepinephrine (NE)-induced lipolysis, the expression of peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) and uncoupling protein 1 (UCP-1), as well as basal mitochondrial respiration in BA. In conclusion, our results have demonstrated for the first time the unique role of brown adipocytes as target cells of sirolimus suggesting that insulin resistance in BAT might play a major role in NODAT development.Peer reviewe