Estudio de la biocompatibilidad in vitro de formulaciones acrílicas autocurables portadoras de bisfosfonatos de nueva síntesis

En este trabajo se presenta el estudio de la biocompatibilidad in vitro, empleando cultivos celulares de fibroblastos embrionarios humanos, de tres sistemas acrílicos de liberación controlada de bisfosfonatos (BFs). Estos sistemas, basados en metacrilato de metilo (MMA), metacrilato de vitamina E (MVE) y trietilenglicol dimetacrilato (TEGDMA), han sido diseñados para su aplicación en el tratamiento de enfermedades caracterizadas por una elevada resorción osteoclástica. En su preparación se ha empleado un BP comercializado (alendronato; ALN) y dos de nueva síntesis, pertenecientes a la segunda y tercera generaciones, el ácido 1-hidroxi-2-[4-aminofenil]etano-1,1-difosfónico (APBP) y el ácido 1-hidroxi-2-[3-indolil]etano-1,1-difosfónico (IBP), respectivamente. La citotoxicidad de MVE ha sido notablemente inferior a la medida para TEGDMA. La citotoxicidad de APBP e IBP se ha comparado con la de ALN, obteniéndose unos valores para la concentración IC50, de15,56, 9,86 y 6,25 mmol/l para APBP, ALN e IBP, respectivamente. Los niveles de citotoxicidad liberada han sido superiores para la formulación portadora de ALN, situándose a continuación los cementos cargados con IBP y APBP. Cuando los cultivos se han establecido sobre la superficie de las formulaciones, se han detectado unos niveles de proliferación celular y adhesión superiores para los sistemas de liberación de IBP y APBP, siendo además inferior en estos casos la mortalidad celular.This paper reports the in vitro biocompatibility study, using human embryonic fibroblast cultures, of three acrylic systems for controlled release of bisphosphonates (BFs). These systems, that are based on methyl methacrylate (MMA), vitamin E methacrylate (MVE) and triethyleneglycol dimethacrylate (TEGDMA), have been designed for its application in the treatment of pathologies characterized by a high osteoclastic resorption. A commercial BP (alendronate; ALN) and two novel BPs, belonging to the second and third generations, 1-hydroxy-2-[4-aminophenyl]ethane-1,1-diphosphonic acid (APBP) and 1-hydroxy-2-[3-indolyl]ethane-1,1-diphosphonic acid (IBP), respectively, have been used in their preparation. The cytotoxicity of MVE has been notably lower than that measured for TEGDMA. Cytotoxicity of APBP and IBP was evaluated along with that of ALN, and the following values of the IC50, concentration were obtained for APBP, ALN and IBP respectively: 15.56, 9.86 and 6.25 mmol/l. The cytotoxicity levels obtained from the extracts of the cured systems were higher for the formulation containing ALN, followed by the cements loaded with IBP and APBP. When the cultures were seeded directly on the formulations surface, higher levels of cellular proliferation and adhesion were obtained for the systems containing IBP and APBP, the cellular mortality being also inferior in these cases

Análisis de la regeneración natural de Pinus pinaster Ait. en los arenales de Almazán-Bayubas (Soria, España)

Natural regeneration is a key factor for the maintenance of forest stands through the time. Research on this process, considering the different forestry treatments to manage them, it is high important for the development of sustainable forestry models. In this study, numerous ecological variables, related to stand, site and seedlings, have been analysed in areas that have been undergone clearcutting leaving seed trees, in Almazán-Bayubas (Soria, Spain). The sampling was systematic using a 100 m squared grid and pots were divided into four quadrants of 4.9 m2 of surface and 2.5 m of radius. After data descriptive analysis, several models were built to predict the regeneration probability, viable density, productivity and rate of height’s growth. Explanatory variables including in were directly related to soil cover, being shrub cover, logging cover and soil sand content the more significant.La regeneración natural es un proceso clave para el mantenimiento de las masas forestales en el tiempo. Analizarlo, teniendo en cuenta los tratamientos selvícolas empleados para gestionar los rodales, es de vital importancia para el desarrollo de modelos de gestión forestal sostenible. En este estudio se han inventariado numerosas variables relativas a la estación, a la masa y al diseminado de zonas en tratamiento de regeneración en la modalidad de corta a hecho en dos tiempos por fajas en rodales de Almazán-Bayubas (Soria). El muestreo está basado en el método de los cuadrantes ocupados y fue sistemático y de malla cuadrada de 100 m de lado. Las parcelas utilizadas fueron circulares, de radio fijo 2,5 m y 4,9 m2 de superficie por cuadrante. Tras el análisis descriptivo de las variables se elaboraron varios modelos para predecir la probabilidad de éxito de regeneración, la densidad de pies viables, la capacidad productiva y el crecimiento medio anual en altura. Las variables explicativas incluidas resultaron estar relacionadas con el tipo de cobertura del suelo, siendo cobertura y altura del matorral total, de los restos de corta y contenido en arena del suelo las más significativas

Ultra-processed food consumption is associated with renal function decline in older adults: A prospective cohort study

Ultra-processed food (UPF) consumption has been associated with increased risk of cardiovascular risk factors and mortality. However, little is known on the UPF effect on renal function. The aim of this study is to assess prospectively the association between consumption of UPF and renal function decline. This is a prospective cohort study of 1312 community-dwelling individuals aged 60 and older recruited during 2008–2010 and followed up to December 2015. At baseline, a validated dietary history was obtained. UPF was identified according to NOVA classification. At baseline and at follow-up, serum creatinine (SCr) and estimated glomerular filtration rate (eGFR) levels were ascertained and changes were calculated. A combined end-point of renal decline was considered: SCr increase or eGFR decreased beyond that expected for age. Logistic regression with adjustment for potential confounders was performed. During follow-up, 183 cases of renal function decline occurred. The fully adjusted odds ratios (95% CI) of renal function decline across terciles of percentage of total energy intake from UPF were 1.56 (1.02–2.38) for the second tercile, and 1.74 (1.14–2.66) for the highest tercile; p-trend was 0.026. High UPF consumption is independently associated with an increase higher than 50% in the risk of renal function decline in Spanish older adults

Research-based flow cytometry assays for pathogenic assessment in the human B-cell biology of gene variants revealed in the diagnosis of inborn errors of immunity: a Bruton’s tyrosine kinase case-study

IntroductionInborn errors of immunity (IEI) are an expanding group of rare diseases whose field has been boosted by next-generation sequencing (NGS), revealing several new entities, accelerating routine diagnoses, expanding the number of atypical presentations and generating uncertainties regarding the pathogenic relevance of several novel variants.MethodsResearch laboratories that diagnose and provide support for IEI require accurate, reproducible and sustainable phenotypic, cellular and molecular functional assays to explore the pathogenic consequences of human leukocyte gene variants and contribute to their assessment. We have implemented a set of advanced flow cytometry-based assays to better dissect human B-cell biology in a translational research laboratory. We illustrate the utility of these techniques for the in-depth characterization of a novel (c.1685G>A, p.R562Q) de novo gene variant predicted as probably pathogenic but with no previous insights into the protein and cellular effects, located in the tyrosine kinase domain of the Bruton’s tyrosine kinase (BTK) gene, in an apparently healthy 14-year-old male patient referred to our clinic for an incidental finding of low immunoglobulin (Ig) M levels with no history of recurrent infections.Results and discussionA phenotypic analysis of bone marrow (BM) revealed a slightly high percentage of pre-B-I subset in BM, with no blockage at this stage, as typically observed in classical X-linked agammaglobulinemia (XLA) patients. The phenotypic analysis in peripheral blood also revealed reduced absolute numbers of B cells, all pre-germinal center maturation stages, together with reduced but detectable numbers of different memory and plasma cell isotypes. The R562Q variant allows Btk expression and normal activation of anti-IgM-induced phosphorylation of Y551 but diminished autophosphorylation at Y223 after anti IgM and CXCL12 stimulation. Lastly, we explored the potential impact of the variant protein for downstream Btk signaling in B cells. Within the canonical nuclear factor kappa B (NF-κB) activation pathway, normal IκBα degradation occurs after CD40L stimulation in patient and control cells. In contrast, disturbed IκBα degradation and reduced calcium ion (Ca2+) influx occurs on anti-IgM stimulation in the patient’s B cells, suggesting an enzymatic impairment of the mutated tyrosine kinase domain

Humanized medium (h7H) allows long-term primary follicular thyroid cultures from human normal thyroid, benign neoplasm, and cancer

Mechanisms of thyroid physiology and cancer are principally studied in follicular cell lines. However, human thyroid cancer lines were found to be heavily contaminated by other sources, and only one supposedly normal-thyroid cell line, immortalized with SV40 antigen, is available. In primary culture, human follicular cultures lose their phenotype after passage. We hypothesized that the loss of the thyroid phenotype could be related to culture conditions in which human cells are grown in medium optimized for rodent culture, including hormones with marked differences in its affinity for the relevant rodent/human receptor.|The objective of the study was to define conditions that allow the proliferation of primary human follicular thyrocytes for many passages without losing phenotype.|Concentrations of hormones, transferrin, iodine, oligoelements, antioxidants, metabolites, and ethanol were adjusted within normal homeostatic human serum ranges. Single cultures were identified by short tandem repeats. Human-rodent interspecies contamination was assessed.|We defined an humanized 7 homeostatic additives medium enabling growth of human thyroid cultures for more than 20 passages maintaining thyrocyte phenotype. Thyrocytes proliferated and were grouped as follicle-like structures; expressed Na+/I- symporter, pendrin, cytokeratins, thyroglobulin, and thyroperoxidase showed iodine-uptake and secreted thyroglobulin and free T3. Using these conditions, we generated a bank of thyroid tumors in culture from normal thyroids, Grave's hyperplasias, benign neoplasms (goiter, adenomas), and carcinomas.|Using appropriate culture conditions is essential for phenotype maintenance in human thyrocytes. The bank of thyroid tumors in culture generated under humanized humanized 7 homeostatic additives culture conditions will provide a much-needed tool to compare similarly growing cells from normal vs pathological origins and thus to elucidate the molecular basis of thyroid disease.Ministerio de Ciencia e InnovaciónInstituto de Salud Carlos IIIXunta de GaliciaFondo Social Europeo of the European Communit

HABs in coastal upwelling systems: Insights from an exceptional red tide of the toxigenic dinoflagellate Alexandrium minutum

Alexandrium minutum blooms generally occur in semi-enclosed sites such as estuaries, harbours and lagoons, where enhanced stratification, restricted circulation and accumulation of resting cysts in the sediment set suitable habitat conditions for the proliferation of this paralytic shellfish poisoning toxigenic species. In the Galician Rías Baixas (NW Iberian Peninsula), according to weekly time-series between 1994 and 2020, blooms of A. minutum were recurrent in small, shallow estuarine bays inside the Rías de Vigo and Pontevedra, but rarely detected, and if so at low concentrations, out of these environments. However, from May to July 2018 it developed as usual in the small inner bays but then spread over both Rías (Vigo and Pontevedra) causing discoloured waters during one month and prolonged harvesting closures. Meteorological conditions during that period (rains / runoff higher than climatological averages, sustained temperature increment and oscillating wind pattern –i.e., series of upwelling-relaxation cycles), fostered optimal circumstances for the development of that extensive and massive proliferation: strong vertical stratification and the alternation of retention and dispersion processes. Simulations from a particle tracking model portrayed the observed bloom development phases: onset and development inside a small inner bay; transport within the surface layer, from these sites towards the interior parts of the Ría; and dispersion all over the embayment. Seedbeds with high concentrations of resting cysts were detected several months after the bloom, which may have favoured flourishment of A. minutum in the following two years, markedly in 2020. The present work contributes to the general understanding of the dynamics of harmful algal blooms (HABs), from which surveillance indicators of the state of marine ecosystems and their evolution can be derived. We hypothesize that the intensity and frequency of A. minutum proliferations in the Galician Rías could increase under projected climate trends.Postprint2,69

Bloom dynamics of an exceptional red tide of the toxigenic dinoflagellate

The toxic dinoflagellate Alexandrium minutum generally proliferates in semi-enclosed sites such as estuaries, harbours and lagoons, where stratification, restricted circulation and accumulation of resting cysts set suitable conditions for its development. In the Galician Rías (NW Iberian Peninsula), its blooms follow also this pattern. They are recurrent in small, shallow estuarine bays inside the Rías, but rarely detected, and if so in minor amount, out of these areas. However, a massive proliferation of A. minutum from June to July 2018 in the Rías Baixas (Vigo and Pontevedra) changed this picture. The bloom initiated in semi-enclosed waters, as previously described for this species, but thereafter spread to the whole embayments where persisted more than one month. It generated a noticeable red tide with disperse patches that became heavily concentrated inside the port of Vigo. During that period shellfish harvesting closures and paralytic shellfish toxins in certain marine invertebrates and fish were reported for the first time in Spain. Meteorological conditions (higher than usual rains/runoff, sustained temperature increment and oscillating wind pattern promoting a series of upwelling-relaxation cycles) fostered optimal circumstances for the outbreak of A. minutum: strong vertical stratification and the alternation of retention and dispersion processes. Simulations from a particle tracking model portrayed the observed bloom development phases: onset, transport within the surface layer towards the interior parts of the Ría of Vigo, and dispersion all over the embayment. High concentrations of resting cysts were detected several months after the bloom, which may have favoured flourish of A. minutum in the following years, markedly in 2020

Food groups consumption and sociodemographic characteristics in Mexican population

Objective. To estimate the recommendable and non-recommendable food groups for usual consumption by sociodemographic characteristics in Mexican population. Materials and methods. Through a 7-day, semi-quan­titative food frequency questionnaire used in 2016 National Health and Nutrition Survey, we estimated the proportions of population (preschool and school children, adolescents and adults) who consumed food groups that are relevant for public health by area, region and socioeconomic status (SES). Results. Less than 50% of population consumed vegetables daily; almost 80% of the population consumed plain water daily and sweetened beverages (3 d/week). Center and Mexico City regions had the highest percent­age of fruits and vegetables consumers (p<0.012). High SES had the highest consumer´s percentage of recommendable and non-recommendable food groups. Conclusions. A high percentage of the population do not consume fruits, vegetables and plain water daily