Cannabidiol skews biased agonism at cannabinoid CB1 and CB2 receptors with smaller effect in CB1-CB2 heteroreceptor complexes

Currently, biased agonism is at the center stage of drug development approaches. We analyzed effects of a battery of cannabinoids plus/minus cannabidiol (CBD) in four functional parameters (cAMP levels, phosphorylation of extracellular signal–regulated kinases (ERK1/2), β-arrestin recruitment and label-free/DMR) in HEK-293T cells expressing cannabinoid receptors, CB or CB, or CB-CB heteroreceptor complexes. In all cases two natural agonists plus two selective synthetic agonists were used. Furthermore, the effect of cannabidiol, at a dose (100 nM) that does not allow significant binding to the orthosteric center of either receptor, was measured. From the huge amount of generated data, we would like to highlight that the two psychotropic molecules (Δ-tetrahydrocannabinol/THC and CP-55940) showed similar bias in CBR and that the bias of THC was particularly relevant toward MAPK pathway. Furthermore, THC did not activate the G protein coupled to CBR. Interestingly, the biased agonism was reduced when assays were performed in cells expressing the two receptors, thus suggesting that the heteromer allows less functional selectivity. In terms of cannabidiol action, the phytocannabinoid altered the functional responses, likely by allosteric means, and modified potency, agonist IC/EC values and biased agonism in qualitative and/or quantitative different ways depending on the agonist. The effect of cannabidiol on anandamide actions on both cannabinoid receptors was particularly noteworthy as was significantly different from that of other compounds. Results are a compendium of data on biased agonism on cannabinoid receptors in the absence and presence of cannabidiol. In addition, for the first time, GPCR biased agonism is characterized in an heteromeric context.This work was partially supported by grants from the Spanish Ministry of Economy and Competitiveness (Ref. no. BFU2015-64405-R and SAF2017-84117-R; they may include FEDER funds) and by grant 201413-30 from: Fundació la Marató de TV3Peer Reviewe

Hydroxyapatite-filled osteoinductive and piezoelectric nanofibers for bone tissue engineering

In this study entitled “Hydroxyapatite-filled osteoinductive and piezoelectric nanofibers for bone tissue engineering”, we describe the development of novel hydroxyapatite (HAp)-filled osteoinductive piezoelectric poly(vinylidene fluoride-cotetrafluoroethylene) (PVDF-TrFE) electrospun nanofibers as a potential strategy for supporting bone repair in delayed-union and non-union osteoporotic-related fractures, for which current clinical techniques have proven to be largely inadequate and scaffold-based tissue engineering approaches hold significant promise. While the piezoelectric properties of native bone tissue have been extensively discussed in the literature, including their key role in preserving tissue homeostasis and promoting tissue repair, they have been widely neglected in the design of scaffolds for bone tissue engineering (BTE) applications. Piezoelectric scaffolds can be used not only for mimicking the native piezoelectric features of bone but also to provide a platform for applying electrical or mechanical stimuli to damaged tissue, contributing to an accelerated regeneration process. The nanofibrous scaffolds generated in this study were capable of replicating the main electrical, structural and compositional properties of bone extracellular matrix (ECM). To the best of our knowledge, this was the first time that the combination of HAp with the piezoelectric polymer PVDF-TrFE was found to induce key shifts in the chemical structure of the polymer and promote ß phase nucleation, not only enhancing the piezoelectric features of the constructs but also improving their surface properties, including their ability to support mineralization in vitro. The HAp nanoparticles also provided meaningful bone-like biological cues (osteoinduction), enhancing the osteogenic differentiation of seeded human mesenchymal stem/stromal cells (hMSCs), which was confirmed by an increased ALP activity, cellderived calcium deposition and expression of important osteogenic gene markers. Overall, our findings highlight, for the first time, the potential of combining PVDFTrFE and HAp for developing electroactive and osteoinductive nanofibrous constructs with improved piezoelectric properties, surface features and osteogenic potential capable of improving bone tissue regeneration.Peer ReviewedPostprint (published version

1873-1874, End of a Century?: Time and Space in Valera's Pepita Jiménez, Ros de Olano's Jornadas de retorno, and Alarcón's El sombrero de tres picos and La Alpujarra

This article argues for the existence of a literature of the first Spanish Republic in the early 1870s. Valera's Pepita Jimenez makes sense in relation to this literature, rather than in comparison with 'Realism'. The literature of the first republic is distinguished by two facets: an ongoing dialogue with Ros de Olano's experiments in simultaneous compression and extension of form; and a belief that the nineteenth-century revolutionary spirit of the age has reached a critical end point, and needs reinvention that leads to Restoration politics

Antibiotic resistance prevalence and trends in patients infected with helicobacter pylori in the period 2013–2020: Results of the european registry on h. pylori management (hp-eureg)

Background: Bacterial antibiotic resistance changes over time depending on multiple factors; therefore, it is essential to monitor the susceptibility trends to reduce the resistance impact on the effectiveness of various treatments. Objective: To conduct a time-trend analysis of Helicobacter pylori resistance to antibiotics in Europe. Methods: The international prospective European Registry on Heli-cobacter pylori Management (Hp-EuReg) collected data on all infected adult patients diagnosed with culture and antimicrobial susceptibility testing positive results that were registered at AEG-REDCap e-CRF until December 2020. Results: Overall, 41,562 patients were included in the Hp-EuReg. Culture and antimicrobial susceptibility testing were performed on gastric biopsies of 3974 (9.5%) patients, of whom 2852 (7%) were naive cases included for analysis. The number of positive cultures decreased by 35% from the period 2013–2016 to 2017–2020. Concerning naïve patients, no antibiotic resistance was found in 48% of the cases. The most frequent resistances were reported against metronidazole (30%), clarithromycin (25%), and levofloxacin (20%), whereas resistances to tetracycline and amoxicillin were below 1%. Dual and triple resistances were found in 13% and 6% of the cases, respectively. A decrease (p < 0.001) in the metronidazole resistance rate was observed between the 2013–2016 (33%) and 2017–2020 (24%) periods. Conclusion: Culture and antimicrobial susceptibility testing for Helicobacter pylori are scarcely performed (<10%) in Europe. In naïve patients, Helicobacter pylori resistance to clarithromycin remained above 15% throughout the period 2013–2020 and resistance to levofloxacin, as well as dual or triple resistances, were high. A progressive decrease in metronidazole resistance was observed

Efecto de la variabilidad de las propiedades físico-químicas del suelo en el rendimiento de la halófita costera Crithmum maritimum L.

Póster presentado en el XV Congreso Nacional de la AEET. "El valor de la naturaleza par una sociedad global"18-21 de octubre, Plasencia, CáceresEl hinojo marino (Crithmum maritimum L.) es una planta halófita perenne perteneciente a la familia Apiaceae, típica de ecosistemas costeros de Europa Occidental. Dada su elevada calidad nutricional para consumo humano, en los últimos años diferentes estudios han resaltado su potencial como cultivo comercial para la agricultura salina, una alternativa sostenible y prometedora ante la creciente salinización de las tierras agrícolas. Sin embargo, existe una falta de información referente a los rangos de tolerancia de la especie frente a los principales factores ambientales ligados a su cultivo, a lo que se suman diversas dificultades prácticas ligadas al cultivo de una especie no domesticada. Además, varios estudios han puesto de manifiesto un alto grado de variabilidad en los rangos de tolerancia a la sal, así como en el perfil nutricional en función de la procedencia geográfica del material. En este estudio se analizó el rendimiento de la especie (en términos de crecimiento, productividad, éxito reproductivo y calidad nutricional) en relación a las propiedades físico-químicas del suelo (pH, conductividad eléctrica, textura, contenido en materia orgánica y macro- y micronutrientes) en siete poblaciones naturales, representativas de los diferentes tipos de hábitat de la especie, localizadas en el sur de la Península Ibérica. Los resultados de este estudio permiten avanzar en el conocimiento ecológico y los mecanismos de tolerancia de las halófitas frente a diferentes factores abióticos, como la salinidad y la riqueza nutricional del suelo, aportando asimismo información relevante de cara a la valorización de la especie para su explotación en agricultura salinaN

Systematic Single-Cell Analysis of Pichia pastoris Reveals Secretory Capacity Limits Productivity

Biopharmaceuticals represent the fastest growing sector of the global pharmaceutical industry. Cost-efficient production of these biologic drugs requires a robust host organism for generating high titers of protein during fermentation. Understanding key cellular processes that limit protein production and secretion is, therefore, essential for rational strain engineering. Here, with single-cell resolution, we systematically analysed the productivity of a series of Pichia pastoris strains that produce different proteins both constitutively and inducibly. We characterized each strain by qPCR, RT-qPCR, microengraving, and imaging cytometry. We then developed a simple mathematical model describing the flux of folded protein through the ER. This combination of single-cell measurements and computational modelling shows that protein trafficking through the secretory machinery is often the rate-limiting step in single-cell production, and strategies to enhance the overall capacity of protein secretion within hosts for the production of heterologous proteins may improve productivity

Differences in nutrient composition of sea fennel (Crithmum maritimum) grown in different habitats and optimally controlled growing conditions

10 páginas.- 1 figura.- 5 tablas.- referenciasCrithmum maritimum L. is an edible halophyte with large potential in human nutrition field. However, it is unclear whether its nutritional value is maintained throughout the contrasting habitats where it commonly grows (cliffs, sandy and rocky beaches) and the nutritional profile of cultivated plants still remains uncertain. In this work, we provided for the first time a comparison of the nutritional profile of C. maritimum across its different type of habitats in the south of Spain and between wild plants and plant material under optimal growing conditions. The protein, amino acids, lipids, fatty acids, minerals composition and phenolic content of plants were analysed. Plants under field conditions exhibited a nutritionally balanced composition (3.8–6.2 g protein/100 g DW, 4.9–7.5 mg lipids/g WW, 3.9–5.0 g Na/100 g DW), with high phenolic content (30.2–48.0 mg/g DW) regardless of the variability of the contrasting habitats. In contrast, under optimal conditions, C. maritimum showed a greater protein and lipid content (10.2 g/100 g DW and 9.6 mg/g WW, respectively), and lower sodium accumulation (1.2 g/100 g DW), allowing a greater consumption of this halophyte without exceeding the daily intake recommendations. Conversely, phenolics were strongly decreased in these plants (6.1 mg/g DW) likely due to the absence of stress factors. © 2021 Elsevier Inc.This work was financially supported by two grants from the Spanish Ministry of Science, Innovation and Universities (RTI2018-099260-A- I00 to J. Cambroll é and RTI2018-099322-B-100 to X. Moreira).Peer reviewe

Detection and elimination of cellular bottlenecks in protein-producing yeasts

Yeasts are efficient cell factories and are commonly used for the production of recombinant proteins for biopharmaceutical and industrial purposes. For such products high levels of correctly folded proteins are needed, which sometimes requires improvement and engineering of the expression system. The article summarizes major breakthroughs that led to the efficient use of yeasts as production platforms and reviews bottlenecks occurring during protein production. Special focus is given to the metabolic impact of protein production. Furthermore, strategies that were shown to enhance secretion of recombinant proteins in different yeast species are presented