Evaluación de un panel de microsatélites para el control de filiación en razas caprinas españolas de aptitud cárnica

Hemos evaluado el potencial de 20 microsatélites para la realización de los controles de filiación (paternidad y/o maternidad) de las razas caprinas españolas de aptitud cárnica. En base a sus condi- ciones técnicas hemos seleccionado nueve para este fin para poder compatibilizar la eficiencia con el coste económico. La mayor parte de los marcadores son de origen bovino. De los marcadores seleccio- nados se han calculado los valores del Contenido de Información Polimórfica (PIC), y las probabilida- des de exclusión (PE), por marcador y conjuntas, de un progenitor falso dado como verdadero a partir de las frecuencias alélicas de 30 individuos no emparentados de cada una de las 6 razas caprinas espa- ñolas de aptitud cárnica estudiadas (un total de 180 individuos): Moncaína, Blanca Andaluza, Negra Serrana, Blanca Celtibérica, Pirenaica y Azpi Gorri. Todos los marcadores seleccionados han sido infor- mativos en estas razas. La probabilidad de exclusión conjunta cuando solo un progenitor es conocido ha oscilado entre el 96,4 % en la raza Moncaína y 98,9 % en la raza Blanca Celtibérica siendo en todos las razas superior al 99% cuando se conocen los dos progenitores y queremos testar si la descendencia está asignada correctamente.Assessment of a microsatellite marker set for parentage testing in six Spanish goat breeds We have analysed 20 microsatellite markers on six Spanish goat populations bred for meat produc- tion. Nine loci were selected for parentage testing due to technical reasons. Polymorphic Informative Content (PIC) and parentage exclusion probabilities per marker and for the whole marker set were computed on allele frequencies from a total of 180 unrelated individuals (30 per breed) belonging to six Spanish goat breeds: Moncaína, Blanca Andaluza, Negra Serrana, Blanca Celtibérica, Pirenaica and Azpi Gorri. The nine markers selected were informative. In order to quantify the usefulness of the microsatellite set for parentage testing, we calculated exclusion probabilities for the two most likely scenarios: a) combined probability of exclusion of a parent when the other is known; and b) combined probability of exclusion when both parent are known and one of them is false. The exclusion proba- bility for the scenario a) varied from 96.4 % (Moncaína breed) to 98,9 % (Blanca Celtibérica breed); the exclusion probabilities for the scenario b) were always higher than 99%Ministerio de Educación y Ciencia RZ01-010-

Estandarización de la técnica de aglutinación directa para el inmunodiagnóstico de la enfermedad de Chagas

Introduction: Chagas’ disease is caused by the parasite Trypanosoma cruzi and its immunological diagnosis is mainly based on the detection of antibodies against T. cruzi using tests such as the ELISA, the indirect fluorescence antibody test (IFAT) and the indirect hemagglutination test (IHAT). The main disadvantage of the IHAT is the need to prepare sheep erythrocytes, whose availability is limited and they have a short duration once prepared. However, there are alternative tests, such as the direct agglutination test (DAT).Objective: To standardize the direct agglutination test for the diagnosis of Chagas disease.Materials and methods: Trypanosoma cruzi epimastigotes were prepared using two protocols, with and without trypsin treatment. The parasites were stained and optimal conditions for parasitic concentration and serum dilutions were determined. We evaluated the technique using sera from patients with Chagas disease, from healthy individuals and from individuals with other parasitic diseases.Results: The optimal parasitic concentration was 500 x 106 parasites/ml using stained parasites without trypsin treatment. The optimal serum dilutions were 1/25, 1/50 y 1/100 and the cut-off point was the 1/50 dilution. The diagnostic indices for the standardized technique were as follows: Sensitivity, 94.3% (95% CI: 79.5-99.0) and specificity, 96.3% (95% CI: 88.8-99.0), with positive and negative predictive values of 91.7% (95% CI: 76.4-97.8) and 97.5% (95% CI: 90.4-99.6), respectively. Cross-reaction was observed only in three sera from individuals with visceral leishmaniasis. The results were compared with those obtained by IHA, ELISA, and IFA, and the concordance rate was 96% and the kappa index, 0.90 (95% CI: 0.81-0.99).Conclusion: The standardized direct agglutination test could be useful for immunodiagnosis of Chagas disease.Introducción. La enfermedad de Chagas es causada por el parásito Trypanosoma cruzi y su diagnóstico inmunológico se basa principalmente en la detección de anticuerpos contra T. cruzi mediante pruebas tales como ELISA, inmunofluorescencia indirecta (IFI) y hemaglutinación indirecta (HAI). Esta última tiene el inconveniente de requerir la preparación de eritrocitos de carnero, difíciles de obtener y de poca duración. Sin embargo, existen pruebas alternativas, como la técnica de aglutinación directa.Objetivo. Estandarizar la técnica de aglutinación directa para el diagnóstico de la enfermedad de Chagas.Materiales y métodos. Se prepararon parásitos epimastigotes de T. cruzi mediante dos protocolos, con tratamiento con tripsina y sin él. Los parásitos se colorearon, y se determinaron las condiciones óptimas de concentración parasitaria y diluciones de suero. Se utilizaron sueros de pacientes con enfermedad de Chagas, de individuos sanos y con otras parasitosis.Resultados. La concentración parasitaria óptima fue de 500 x106 parásitos/ml, utilizando parásitos coloreados y sin tratamiento con tripsina. Las diluciones de suero óptimas fueron de 1/25, 1/50 y1/100, y el punto de corte, la dilución de 1/50. La técnica estandarizada mostró índices diagnósticos de sensibilidad de 94,3 % (IC95% 79,5-99,0) y de especificidad de 96,3 % (IC95% 88,8-99,0); se encontró reacción cruzada en tres sueros de individuos con leishmaniasis visceral, con valores pronósticos positivo y negativo de 91,7 % (IC95% 76,4-97,8) y de 97,5 % (IC95% 90,4-99,6), respectivamente. Se compararon los resultados con los obtenidos por HAI, ELISA e IFI y la concordancia fue de 96 % con un índice kappa de 0,90 (IC95% 0,81-0,99).Conclusión. La técnica de aglutinación directa estandarizada podría ser útil para el inmunodiagnóstico de la enfermedad de Chagas

Protumorigenic effects of Snail-expression fibroblasts on colon cancer cells

et al.Snail1 is a transcriptional factor that plays an important role in epithelial–mesenchymal transition and in the acquisition of invasive properties by epithelial cells. In colon tumors, Snail1 expression in the stroma correlates with lower specific survival of cancer patients. However, the role(s) of Snail1 expression in stroma and its association with patients' survival have not been determined. We used human primary carcinoma-associated fibroblasts (CAFs) or normal fibroblasts (NFs) and fibroblast cell lines to analyze the effects of Snail1 expression on the protumorigenic capabilities in colon cancer cells. Snail1 expression was higher in CAFs than in NFs and, as well as α-SMA, a classic marker of activated CAFs. Moreover, in tumor samples from 50 colon cancer patients, SNAI1 expression was associated with expression of other CAF markers, such as α-SMA and fibroblast activation protein. Interestingly, coculture of CAFs with colon cells induced a significant increase in epithelial cell migration and proliferation, which was associated with endogenous SNAI1 expression levels. Ectopic manipulation of Snail1 in fibroblasts demonstrated that Snail1 expression controlled migration as well as proliferation of cocultured colon cancer cells in a paracrine manner. Furthermore, expression of Snail1 in fibroblasts was required for the coadjuvant effect of these cells on colon cancer cell growth and invasion when coxenografted in nude mice. Finally, cytokine profile changes, particularly MCP-3 expression, in fibroblasts are put forward as mediators of Snail1-derived effects on colon tumor cell migration. In summary, these studies demonstrate that Snail1 is necessary for the protumorigenic effects of fibroblasts on colon cancer cells.This research was supported by the PI12/02037, Fundación Científica AECC, SAF2010-20750, S2010/BMD-2344, RTICC-RD12/0036/0041 and by the Fundación Banco Santander. Antonio García de Herreros’ laboratory was supported by RTICC-RD12/0036/0005 and SAF 2010-16089. Ma Jesús Larriba’s laboratory was supported by RD12/0036/0021. Cristina Peña and José Miguel García are recipients of Miguel Servet Contracts from the Instituto de Salud Carlos III.Peer reviewe

Methodology of the Virtual Reconstruction of Arquitectonic Heritage: Ambassador Vich's Palace in Valencia

The 19th century was disastrous as far as the conservation of architectonic heritage is concerned. The awareness of the importance of preserving monuments that has prevailed since the end of the last century was dazzlingly absent in the previous, leading both to the disappearance of representative heritage works and the plundering of many others. The present study establishes the methodological basis to proceed with the virtual reconstruction of many disappeared architectures, representative of emblematic architectonic typologies. A method based on the combination of deduction and induction allows benchmarks to be created that signify a starting point to which the key and specific elements of each building are later incorporated, from the data extracted from the conserved parts and the graphic, literary and archive documents. The result is the virtual recovery of the general outlines of the architecture: morphology of the plot, volumetry, exterior and interior facades, and the functional layout. The good results obtained in the study of the disappeared Ambassador Vich's Palace, allow the methodology to be extended to the analysis of other similar examples, serving investigators as a tool to carry out an arduous task of deciphering a trail that is increasingly fading with the passing of time.Galiana Agullo, M.; Mas Tomas, MDLA.; Lerma Elvira, C.; Peñalver Martínez, MJ.; Conesa Tejada, S. (2014). Methodology of the Virtual Reconstruction of Arquitectonic Heritage: Ambassador Vich's Palace in Valencia. International Journal of Architectural Heritage. 8(1):94-123. doi:10.1080/15583058.2012.672623S9412381Boix, V. 1979.Historical and topographic Valencia[in Spanish]. Vol. I261 S. A. Printing J. Rius.Estaban Chapapría, J. (2001). Impostación del patio del Embajador Vich en el ex-convento del Carmen (Valencia). Loggia, Arquitectura & Restauración, (12), 26. doi:10.4995/loggia.2001.3605Morrish, S. W., & Laefer, D. F. (2010). Web-Enabling of Architectural Heritage Inventories. International Journal of Architectural Heritage, 4(1), 16-37. doi:10.1080/15583050902731056Lotz, W. 1995.Architecture in Italy 1500–1600 [in Italian]35–37. ed. RizzoliYale University Press.Lourenço, P. B., Peña, F., & Amado, M. (2010). A Document Management System for the Conservation of Cultural Heritage Buildings. International Journal of Architectural Heritage, 5(1), 101-121. doi:10.1080/15583050903318382Vila Ferrer, S. (2001). La recuperación del patio del palacio del Embajador Vich (Valencia). Loggia, Arquitectura & Restauración, (12), 44. doi:10.4995/loggia.2001.3606Zonta, D., Pozzi, M., & Zanon, P. (2008). Managing the Historical Heritage Using Distributed Technologies. International Journal of Architectural Heritage, 2(3), 200-225. doi:10.1080/1558305080206369

What's pain like?

La utilización de la terminología médica en inglés es vital para el desarrollo de las actividades docentes de los estudiantes de 4to. y 5to. año de Medicina y, sobre todo, en las actividades de Educación para el trabajo  que están estipuladas en el programa de las asignaturas Inglés VII, VIII, IX y X.Dentro de ella se encuentra la descripción de la sensación de dolor que incluye un amplio vocabulario que no aparece en los libros de texto de inglés, por lo cual realizamos una revisión minuciosa del tema.Este glosario contiene cincuenta y cinco términos actualizados, asociados con la sensación de dolor y sus equivalentes en Español, así como sesenta y ocho enfermedades o entidades, en las que aparece este léxico. Esta terminología es necesaria para los estudiantes como parte de su entrenamiento docente en inglés, considerando los objetivos de salida del médico general básico.Palabras claves: Dolor, Educación en el trabajo.</span

Lymphocyte Profile and Immune Checkpoint Expression in Drug-Induced Liver Injury: An Immunophenotyping Study

The identification of specific HLA risk alleles in drug-induced liver injury (DILI) points toward an important role of the adaptive immune system in DILI development. In this study, we aimed to corroborate the role of an adaptive immune response in DILI through immunophenotyping of leukocyte populations and immune checkpoint expressions. Blood samples were collected from adjudicated DILI (n = 12), acute viral hepatitis (VH; n = 13), acute autoimmune hepatitis (AIH; n = 9), and acute liver injury of unknown etiology (n = 15) at day 1 (recognition), day 7, and day >30. Blood samples from patients with nonalcoholic fatty liver disease (NAFLD; n = 20) and healthy liver controls (HLCs; n = 54) were extracted at one time point. Leukocyte populations and immune checkpoint expressions were determined based on cell surface receptors, except for CTLA-4 that was determined intracellularly, using flow cytometry. At recognition, DILI demonstrated significantly higher levels of activated helper T-cell (P < 0.0001), activated cytotoxic T-cells (P = 0.0003), Th1 (P = 0.0358), intracellular CTLA-4 level in helper T-cells (P = 0.0192), and PD-L1 presenting monocytes (P = 0.0452) than HLC. These levels approached those of HLC over time. No significant differences were found between DILI and VH. However, DILI presented higher level of activated helper T-cells and CTLA-4 than NAFLD and lower PD-L1 level than AIH. Our findings suggest that an adaptive immune response is involved in DILI in which activated CD4+ and CD8+ play an important role. Increased expression of negative immune checkpoints is likely the effect of peripheral tolerance regulation.The present study has been supported by grants of Instituto de Salud Carlos III cofounded by Fondo Europeo de Desarrollo Regional – FEDER (contract numbers: PI19/00883, PI16/01748, P18-RT-3364-2020, and PT20/000127). CIBERehd and Plataforma ISCiii Ensayos Clínicos are funded by Instituto de Salud Carlos III. Funding for open access charge: Universidad de Málaga/CBUA. The funding sources had no involvement in the study design; in the collection, analysis, and interpretation of data; in the writing of the report, or in the decision to submit the manuscript for publication

Extracellular vesicles from pristane-treated CD38-deficient mice express an antiinflammatory neutrophil protein signature, which reflects the mild lupus severity elicited in these mice

In CD38-deficient (Cd38-/-) mice intraperitoneal injection of pristane induces a lupus-like disease, which is milder than that induced in WT mice, showing significant differences in the inflammatory and autoimmune processes triggered by pristane. Extracellular vesicles (EV) are present in all body fluids. Shed by cells, their molecular make-up reflects that of their cell of origin and/or tissue pathological situation. The aim of this study was to analyze the protein composition, protein abundance, and functional clustering of EV released by peritoneal exudate cells (PECs) in the pristane experimental lupus model, to identify predictive or diagnostic biomarkers that might discriminate the autoimmune process in lupus from inflammatory reactions and/or normal physiological processes. In this study, thanks to an extensive proteomic analysis and powerful bioinformatics software, distinct EV subtypes were identified in the peritoneal exudates of pristane-treated mice: 1) small EV enriched in the tetraspanin CD63 and CD9, which are likely of exosomal origin; 2) small EV enriched in CD47 and CD9, which are also enriched in plasma-membrane, membrane-associated proteins, with an ectosomal origin; 3) small EV enriched in keratins, ECM proteins, complement/coagulation proteins, fibrin clot formation proteins, and endopetidase inhibitor proteins. This enrichment may have an inflammation-mediated mesothelial-tomesenchymal transition origin, representing a protein corona on the surface of peritoneal exudate EV; 4) HDL-enriched lipoprotein particles. Quantitative proteomic analysis allowed us to identify an anti-inflammatory, Annexin A1- enriched pro-resolving, neutrophil protein signature, which was more prominent in EV from pristane-treated Cd38-/- mice, and quantitative differences in the protein cargo of the ECM-enriched EV from Cd38-/- vs WT mice. These differences are likely to be related with the distinct inflammatory outcome shown by Cd38-/- vs WT mice in response to pristane treatment. Our results demonstrate the power of a hypothesis-free and data-driven approach to transform the heterogeneity of the peritoneal exudate EV from pristanetreated mice in valuable information about the relative proportion of different EV in a given sample and to identify potential protein markers specific for the different small EV subtypes, in particular those proteins defining EV involved in the resolution phase of chronic inflammation.Proyecto del plan estatal, Ministerio de Ciencia e Innovacion PT13/0001/011CSIC PT17/0019/0010 PID2020-119567RB-I0

Phenotypic differentiation among native, expansive and introduced populations influences invasion success

Aim: Humans influence species distributions by modifying the environment and by dispersing species beyond their natural ranges. Populations of species that have established in disjunct regions of the world may exhibit trait differentiation from native populations due to founder effects and adaptations to selection pressures in each distributional region. We compared multiple native, expansive and introduced populations of a single species across the world, considering the influence of environmental stressors and transgenerational effects. Location: United States Gulf and Atlantic coasts, United States interior, European Atlantic and Mediterranean coasts, east coast of Australia. Taxon: Baccharis halimifolia L. (eastern baccharis). Methods: We monitored seed germination, seedling emergence, survival and early growth in a common garden experiment, conducted with over 18,200 seeds from 80 populations. We also evaluated the influence of environmental stress and maternal traits on progeny performance. Results: Introduced European Atlantic populations had faster germination and early growth than native populations. However, this was not the case for the more recently naturalized European Mediterranean populations. Introduced Australian populations grew faster than native populations in non-saline environments but had lower survival in saline conditions commonly encountered in the native range. Similarly, expansive inland US populations germinated faster than coastal native populations in non-saline environments but grew and germinated more slowly in saline environments. Maternal inflorescence and plant size were positively related with seed germination and seedling survival, whereas flower abundance was positively correlated with seedling early growth and survival. However, maternal traits explained a much lower fraction of the total variation in early demographic stages of B. halimifolia than did distributional range. Main conclusions: Phenotypic differentiation could allow B. halimifolia to adapt to different biotic and abiotic selection pressures found in each distributional range, potentially contributing to its success in introduced and expansive ranges

Extrusion Cooking Effect on Carbohydrate Fraction in Novel Gluten-Free Flours Based on Chickpea and Rice

Extrusion cooking allows the development of value-added products from pulses, such as gluten-free snacks with added functional properties. The main objective of this study was to evaluate the changes induced by the extrusion process on the carbohydrate fraction (total carbohydrates, soluble sugars and oligosaccharides, dietary fiber, and arabinoxylans) of novel flour formulations based on chickpeas and rice enriched with different dietary fiber sources. Moreover, the influence of the addition of fiber-rich ingredients, such as Fibersol® and passion fruit, on the analyzed compounds was also evaluated. Sucrose was the main soluble sugar found in analyzed formulations, and raffinose was the prevalent oligosaccharide, followed by stachyose. The content of total α-galactosides tended to be higher after extrusion cooking. As a consequence of the extrusion treatment, the content of total and soluble dietary fiber was statistically increased in most of the analyzed samples. In general, no significant changes were observed in total arabinoxylan content as a consequence of the extrusion process, while the content of water-soluble arabinoxylans was significantly increased in extruded formulations. It was observed that the content of total available carbohydrates, stachyose, and water-soluble arabinoxylans were significantly influenced by the addition of passion fruit, Fibersol®, and both. The incorporation of these ingredients in gluten-free formulations based on chickpeas and rice allows one to obtain suitable functional formulations for the development of innovative, gluten-free, extruded snack-type products, which could be an interesting alternative for people with celiac disease