Thrombospondin-1/CD47 interaction regulates Th17 and treg differentiation in psoriasis

Accumulating evidence on the role of Thrombospondin-1 (TSP-1) in the immune response has emerged during the last years. In spite of the importance of TSP-1 not only as anti-angiogenic factor but also as an immunomodulatory molecule, studies on the role of TSP-1 in psoriasis have been neglected. TSP-1 and CD47 expression were analyzed in skin samples from psoriasis patients and control subjects using RT-PCR and immunofluorescence. Expression of these molecules was also evaluated in peripheral blood CD4+ T cells, moDCs, and circulating primary DCs. The functional role of TSP-1/CD47 signaling axis in psoriasis was assessed in Th17 and Treg differentiation assays. Additionally, small interfering RNA assays specific to TSP-1 were performed in CD4+ T cells and monocyte derived DC to specifically evaluate the function of this protein. Lesional skin of psoriasis patients expressed lower TSP-1 and CD47 mRNA levels compared to non-lesional skin or skin from controls. Immunofluorescence staining revealed decreased expression of CD47 in CD45+ dermal cells from psoriasis samples compared to control subjects. Peripheral CD4+ T cells and circulating primary DCs from psoriasis also expressed lower levels of CD47 compared to controls. Although no significant differences were detected in TSP-1 expression in CD4+ T cells and moDCs between patients and controls, TSP-1 expression in psoriasis patients inversely correlated with disease activity evaluated by the Psoriasis Area and Index Activity. Furthermore, exogenous TSP-1 inhibited Th17 differentiation and stimulated the differentiation of CD4+ T cells toward Treg cells. Furthermore, RNA interference specific for TSP-1 confirmed the role of this molecule as a negative regulator of T cell activation. Because of the impact of TSP-1/CD47 signaling axis in Th17 and Treg differentiation, a dysregulated expression of these molecules in the immune cells from psoriasis patients may favor the exacerbated inflammatory response in this diseaseInstituto de Salud Carlos III (AES 2017): PI17/01972 to ED. Janssen; Spanish Ministry of Economy and Competitiveness (MINECO): Plan Nacional de Salud SAF2017-82886-R, Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares (CIBERCV); Proyecto Integrado de Excelencia PIE13/00041, Instituto de Salud Carlos III to FS-M, Instituto de Salud Carlos III PI16/02166, Universidad Autónoma de Madrid-Banco Santander (grant 2017/EEUU/03), and Red Temática de Excelencia en Investigación en Hipoxia (SAF 2017-90794-REDT) to MJC. This research has been co-financed by Fondo Europeo de Desarrollo Regional (FEDER

Evaluación Estructural Mediante Ultrasonidos del Efecto del Espesor de la Junta de Mortero en Muros de Fábrica de Ladrillo Cerámino Sometidos a Esfuerzo de Compresión hasta su Rotura.

En este trabajo se analiza la influencia del espesor de la junta de mortero en el comportamiento estructural de probetas de fábrica de ladrillo cerámico bajo esfuerzos de compresión uniaxial. El ensayo de compresión reproduce la forma habitual de trabajo de estas estructuras. Se han ensayado nueve muros y nueve pilares, combinando tres espesores de junta de mortero, sometidos a carga de compresión hasta rotura. Con el fin de evaluar el progresivo agrietamiento del material durante el proceso de carga, se han realizado medidas con ultrasonidos. Además se han realizado medidas con extensómetros y esclerómetro para contrastar la validez de las medidas de ultrasonidos. Los resultados obtenidos muestran que existe una correlación entre los valores de la fuerza de compresión que soportan las probetas y el tamaño del espesor de la junta: a menor espesor de junta mayor carga de rotura. Del estudio también se desprende que esta relación es más clara en los muros que en los pilares, ya que la esbeltez de los muros es mayor y aparecen efectos de pandeo. La medida con ultrasonidos muestra una buena correlación con las medidas extenso métricas y permite una eficaz detección del agrietamiento interior del material durante el proceso de rotura

pVHL-mediated regulation of the anti-angiogenic protein thrombospondin-1 decreases migration of Clear Cell Renal Carcinoma Cell Lines

Thrombospondin-1 (TSP-1) is a multifunctional matrix protein with antitumor activities due in part to its ability to inhibit angiogenesis, which in turn contributes to determine the fate of many tumours. Previous studies have shown that TSP-1 expression supports normal kidney angiostasis, and decreased TSP-1 levels contribute to the angiogenic phenotype of renal cell carcinomas (RCC). The loss of the von Hippel-Lindau tumour suppressor gene (VHL) in these tumours favours stabilization of the Hypoxia Inducible Factors (HIF), which in turn contribute to adapt tumour cells to hostile environments promoting tumour progression. However, HIF-independent regulation of certain genes might also be involved. We have previously shown that TSP-1 is regulated in hypoxia in clear cell RCC (ccRCC) in a HIF-independent manner; however, the effect of VHL protein (pVHL) on TSP-1 expression has not been evaluated. Our results proved that pVHL loss or mutation in its alpha or beta domain significantly decreased TSP-1 levels in ccRCC in a HIF-independent manner. Furthermore, this regulation proved to be important for ccRCC cells behaviour showing that decreased TSP-1 levels rendered ccRCC cells more migratory. This data substantiates a unique regulation pattern for TSP-1 in a pVHL-dependent manner, which may be relevant in the aggressiveness of ccRCC.This work was supported by grants from the Spanish Government (co-funded by European Regional Development Fund, ERDF/FEDER); PI16/02166 and 2017/EEUU/03 to MJC; Red Temática de Excelencia en Investigación en Hipoxia (SAF 2017-90794-REDT) to MJ

Straightforward purification method for the determination of the activity of glucose oxidase and catalase in honey by extracting polyphenols with a film-shaped polymer

Glucose oxidase (GOX) and catalase (CAT) regulate the amount of H2O2 in honey, by generating or consuming it, so they are related to the antibacterial and antioxidant activity of honey. However, their activities are hardly analysed, since the process requires a previous dialysis that is non-selective, very time-consuming (>24 h), eco-unfriendly (>6L of buffer) and expensive. This research shows the design and performance of a material that selectively removes the actual interferents. The film-shaped-polymer is immersed for 90́ within a honey solution (12.5 mL of buffer), where it interacts exclusively with 1,2-dihydroxybenzenes, which we proved to be the real interferents (the material contains motifs derived from phenylboronic acid to interact with 1,2-diols). Polymeric chains favour condensation to occur exclusively with 1,2-dihydroxybenzenes, excluding monosaccharides. The interferents’ removal using our designed polymer is selective, low cost (1.42€ per test), rapid and eco-friendly (saves 6L of buffer and 20.5 h of experimental workout per sample).We gratefully acknowledge the financial support provided by all funders. Author Jose Miguel García received grant PID2020-113264RB-I00 / AEI / 10.13039/501100011033 funded by MCIN/AEI/ 10.13039/501100011033 and by “ERDF A way of making Europe”. Ana Arnaiz received funding from Ministerio de Universidades-European Union in the frame of NextGenerationEU RD 289/2021 (Universidad Politécnica de Madrid). We also gratefully acknowledge European Regional Development Fund (ERDF). Gianluca Utzeri thanks Fundação para a Ciência e a Tecnologia (FCT, Portugal) for PhD grant (SFR/BD/146358/2019). The Coimbra Chemistry Centre is supported by the FCT, through Projects UIDB/00313/2020 and UIDP/00313/2020. To all the beekeepers who provided a sample of honey for this study

Photoferroelectric thin films for flexible systems by a three-in-one solution-based approach

The effective incorporation of (multi)functional oxides into next-generation flexible electronics systems requires novel fabrication technologies that enable the direct integration of crystalline oxide layers in them. Unfortunately, this is considerably challenging due to the thermal incompatibility between the crystallization temperatures of metal oxides (>600 degrees C) and the thermal stability of the flexible polymer substrates conventionally used (<400 degrees C). Here, it is shown that BiFeO(3)thin films can be grown on flexible plastic by solution processing involving three different but complementary strategies to induce the crystallization of the perovskite phase at a lower temperature limit of 325 degrees C. This "three-in-one" approach is based on the synthesis of tailored metal precursors i) with a molecular structure resembling the crystalline structure of the oxide phase, which additionally allows both ii) photochemical and iii) internal combustion reactions taking place in the thin films. The flexible BiFeO(3)thin films obtained from a specifically designed molecular complex withN-methyldiethanolamine yield a large remnant polarization of 17.5 mu C cm(-2), also showing photovoltaic and photocatalytic effects. This result paves the way for the direct integration of an interesting class of oxides with photoferroelectric properties in flexible devices with multiple applications in information and communication technology, and energy

Antiretroviral therapy duration and immunometabolic state determine efficacy of ex vivo dendritic cell-based treatment restoring functional HIV-specific CD8+ T cells in people living with HIV.

Dysfunction of CD8+ T cells in people living with HIV-1 (PLWH) receiving anti-retroviral therapy (ART) has restricted the efficacy of dendritic cell (DC)-based immunotherapies against HIV-1. Heterogeneous immune exhaustion and metabolic states of CD8+ T cells might differentially associate with dysfunction. However, specific parameters associated to functional restoration of CD8+ T cells after DC treatment have not been investigated. We studied association of restoration of functional HIV-1-specific CD8+ T cell responses after stimulation with Gag-adjuvant-primed DC with ART duration, exhaustion, metabolic and memory cell subsets profiles. HIV-1-specific CD8+ T cell responses from a larger proportion of PLWH on long-term ART (more than 10 years; LT-ARTp) improved polyfunctionality and capacity to eliminate autologous p24+ infected CD4+ T cells in vitro. In contrast, functional improvement of CD8+ T cells from PLWH on short-term ART (less than a decade; ST-ARTp) after DC treatment was limited. This was associated with lower frequencies of central memory CD8+ T cells, increased co-expression of PD1 and TIGIT and reduced mitochondrial respiration and glycolysis induction upon TCR activation. In contrast, CD8+ T cells from LT-ARTp showed increased frequencies of TIM3+ PD1- cells and preserved induction of glycolysis. Treatment of dysfunctional CD8+ T cells from ST-ARTp with combined anti-PD1 and anti-TIGIT antibodies plus a glycolysis promoting drug restored their ability to eliminate infected CD4+ T cells. Together, our study identifies specific immunometabolic parameters for different PLWH subgroups potentially useful for future personalized DC-based HIV-1 vaccines. NIH (R21AI140930), MINECO/FEDER RETOS (RTI2018-097485-A-I00) and CIBERINF grants.NIH (R21AI140930), MINECO/FEDER RETOS (RTI2018-097485-A-I00) and CIBERINF grants. We would like to thank the NIH AIDS Reagent Pro- gram, Division of AIDS, NIAID, NIH for providing HIV-1 PTE Gag Peptide Pool from NIAID, DAIDS (cat #11057) for the study. We would also like to thank Alvaro Serrano Navarro, for his help on adapting the lin- ear mixed model previously described by Martin- C ofreces N. et al83 to our data. Graphical schematic rep- resentations were created with BioRender.com. EMG was supported by the NIH R21 program (R21AI140930), the Ramón y Cajal Program (RYC2018- 024374-I), the MINECO/FEDER RETOS program (RTI2018-097485-A-I00), by Comunidad de Madrid Talento Program (2017-T1/BMD-5396) and by Gilead becas de investigaci on (GLD19/00168). EMG and IDS are supported by Centro de Investigación Biomédica en Red (CIBERINF) de Enfermedades Infecciosas (CB21/ 13/00107). MCM was supported by NIH R21 program (R21AI140930), “La Caixa Banking Foundation (H20- 00218) and Gilead becas de investigaci on (GLD19/ 00168). MJB is supported by the Miguel Servet program funded by the Spanish Health Institute Carlos III (CP17/00179), the MINECO/FEDER RETOS program (RTI2018-101082-B-100), and Fundació La Marat o TV3 (201805-10FMTV3). EMG and MJB are both funded by “La Caixa Banking Foundation (H20-00218) and by REDINCOV grant from Fundació La Marat o TV3. FSM was supported by SAF2017-82886-R and PDI-2020- 120412RB-I00 grants from the Ministerio de Ciencia e Innovaci on, and HR17-00016 grant from “La Caixa Banking Foundation. HF was funded by PI21/01583 grant from Ministerio de Ciencia e Innovación, Instituto de Salud Carlos III. MJC was supported by PID2019- 104406RB-I00 from Ministerio de Ciencia e Innovación. ISC was funded by the CM21/00157 Rio- Hortega grant. IT was supported by grant for the pro- motion of research studies master-UAM 2021.S

Modified magnetic anisotropy at LaCoO_(3)/La_(0.7)Sr_(0.3)MnO_(3) interfaces

Controlling magnetic anisotropy is an important objective towards engineering novel magnetic device concepts in oxide electronics. In thin film manganites, magnetic anisotropy is weak and it is primarily determined by the substrate, through induced structural distortions resulting from epitaxial mismatch strain. On the other hand, in cobaltites, with a stronger spin orbit interaction, magnetic anisotropy is typically much stronger. In this paper, we show that interfacing La0.7Sr0.3MnO3 (LSMO) with an ultrathin LaCoO3 (LCO) layer drastically modifies the magnetic anisotropy of the manganite, making it independent of the substrate and closer to the magnetic isotropy characterizing its rhombohedral structure. Ferromagnetic resonance measurements evidence a tendency of manganite magnetic moments to point out-of-plane suggesting non collinear magnetic interactions at the interface. These results may be of interest for the design of oxide interfaces with tailored magnetic structures for new oxide devices

T908 polymeric micelles improved the uptake of Sgc8-c aptamer probe in tumor-bearing mice: a co-association study between the probe and preformed nanostructures

Aptamers are oligonucleotides that have the characteristic of recognizing a target with high affinity and specificity. Based on our previous studies, the aptamer probe Sgc8-c-Alexa647 is a promising tool for molecular imaging of PTK7, which is an interesting biomarker in cancer. In order to improve the delivery of this probe as well as create a novel drug delivery nanosystem targeted to the PTK7 receptor, we evaluate the co-association between the probe and preformed nanostructures. In this work, preformed pegylated liposomes (PPL) and linear and branched pristine polymeric micelles (PMs), based on PEO–PPO–PEO triblock copolymers were used: poloxamer F127® and poloxamines T1307® and T908®. For it, Sgc8-c-Alexa647 and its co-association with the different nanostructures was exhaustively analyzed. DLS analysis showed nanometric sizes, and TEM and AFM showed notable differences between free- and co-associated probe. Likewise, all nanosystems were evaluated on A20 lymphoma cell line overexpressing PTK7, and the confocal microscopy images showed distinctness in cellular uptake. Finally, the biodistribution in BALB/c mice bearing lymphoma-tumor and pharmacokinetic study revealed an encouraging profile for T908-probe. All data obtained from this work suggested that PMs and, more specifically T908 ones, are good candidates to improve the pharmacokinetics and the tumor uptake of aptamer-based probes

Photochemical solution processing of films of metastable phases for flexible devices: the beta-Bi2O3 polymorph

The potential of UV-light for the photochemical synthesis and stabilization of non-equilibrium crystalline phases in thin films is demonstrated for the beta-Bi2O3 polymorph. The pure beta-Bi2O3 phase is thermodynamically stable at high temperature (450-667 degrees C), which limits its applications in devices. Here, a tailored UV-absorbing bismuth(III)-N-methyldiethanolamine complex is selected as an ideal precursor for this phase, in order to induce under UV-light the formation of a -Bi-O-Bi- continuous network in the deposited layers and the further conversion into the beta-Bi2O3 polymorph at a temperature as low as 250 degrees C. The stabilization of the beta-Bi2O3 films is confirmed by their conductivity behavior and a thorough characterization of their crystal structure. This is also supported by their remarkable photocatalytic activity. Besides, this processing method has allowed us for the first time the preparation of beta-Bi2O3 films on flexible plastic substrates, which opens new opportunities for using these materials in potential applications not available until now (e.g., flexible photocatalytic reactors, self-cleaning surfaces or wearable antimicrobial fabrics). Therefore, photochemical solution deposition (PCSD) demonstrates to be not only an efficient approach for the low temperature processing of oxide films, but also an excellent alternative for the stabilization of metastable phases

Hypoxia Negatively Regulates Antimetastatic PEDF in Melanoma Cells by a Hypoxia Inducible Factor-Independent, Autophagy Dependent Mechanism

Pigment epithelium-derived factor (PEDF), a member of the serine protease inhibitor (SERPIN) superfamily, displays a potent antiangiogenic and antimetastatic activity in a broad range of tumor types. Melanocytes and low aggressive melanoma cells secrete high levels of PEDF, while its expression is lost in highly aggressive melanomas. PEDF efficiently abrogates a number of functional properties critical for the acquisition of metastatic ability by melanoma cells, such as neovascularization, proliferation, migration, invasiveness and extravasation. In this study, we identify hypoxia as a relevant negative regulator of PEDF in melanocytes and low aggressive melanoma cells. PEDF was regulated at the protein level. Importantly, although downregulation of PEDF was induced by inhibition of 2-oxoglutarate-dependent dioxygenases, it was independent of the hypoxia inducible factor (HIF), a key mediator of the adaptation to hypoxia. Decreased PEDF protein was not mediated by inhibition of translation through untranslated regions (UTRs) in melanoma cells. Degradation by metalloproteinases, implicated on PEDF degradation in retinal pigment epithelial cells, or by the proteasome, was also excluded as regulatory mechanism in melanoma cells. Instead, we found that degradation by autophagy was critical for PEDF downregulation under hypoxia in human melanoma cells. Our findings show that hypoxic conditions encountered during primary melanoma growth downregulate antiangiogenic and antimetastasic PEDF by a posttranslational mechanism involving degradation by autophagy and could therefore contribute to the acquisition of highly metastatic potential characteristic of aggressive melanoma cells