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En la actualidad el uso y aprovechamiento de los recursos vegetales ha derivado en numerosas investigaciones etnobotánicas, las cuales vienen documentando una rápida erosión de conocimientos tradicionales en las culturas nativas peruanas, incentivando su preservación y consecuente revalorización. Con el fin de facilitar el análisis de la data obtenida en los múltiples estudios realizados, esta investigación constituye una aproximación de los criterios de clasificación de las categorías de uso de las especies vegetales asociadas a la vida y cultura del poblador peruano. La definición de las categorías de uso presentes en este documento constituye el resultado de 130 expediciones de campo realizadas por los autores en comunidades costeras, andinas y amazónicas peruanas, así como en la evaluación de 17 281 registros de usos documentados en nueve herbarios peruanos y extranjeros. Se proponen nueve categorías de usos, subdivididas en 60 subcategorías: medicinal (21), social (8), alimento humano (3), alimento para animales (3), materiales (10), tóxicas (4), etnoveterinario (2), ambiental (6) y combustible (3). Se concluye que la propuesta de una estandarización de categorías de uso facilitará la compilación y análisis del uso de plantas dentro de una comunidad, y a la vez permitirá realizar comparaciones entre ecosistemas y/o comunidades. Nowadays, the use and harnessing of vegetal resources has resulted in much ethnobotanical research, which has documented a rapid erosion of traditional knowledge in native Peruvian cultures, encouraging its preservation and consequent revaluation. In order to ease the analysis of the data obtained in the multiple studies that have been made, this proposal constitutes an approximation of the classification criteria for the categories of plant uses associated with the life and culture of Peruvian inhabitants. These use categories’ definitions are the result of 130 field expeditions carried out by the authors, in coastal, Andean and Amazon Peruvian communities, as well as the evaluation of 17 281-use registries documented in nine foreign and Peruvian herbaria. The nine use categories proposed in this article, are divided in 60 subcategories: medicinal (21), social (8), human food (3), animal food (3), materials (10), toxic (4), ethnoveterinary medicine (2), environmental (6) and fuel (3). The authors conclude that the proposal of the use categories standardization will simplify the compilation and analysis of plant uses in a community, and it will allow making comparisons between different ecosystems and communities.

Las industrias pesqueras y de acuicultura generan, cada año, un conjunto de residuos o desechos que incluyen piel, cabeza, vísceras, recortes y espinazos, representando más del 60% en volumen productivo. Estos residuos tienen altos contenidos de proteínas, y normalmente son procesados en productos de bajo valor comercial, como alimentos para animales, harina de residuos y fertilizantes. En los últimos años, se han venido desarrollando tecnologías para el aprovechamiento de éstos residuos y convertirlos en bioproductos de mayor valor agregado, como son los hidrolizados de proteínas, con interesantes aplicaciones en la alimentación animal. Los hidrolizados proteicos de pescado son productos obtenidos de la degradación enzimática o química de las proteínas de pescado en péptidos más pequeños, aminoácidos libres y nucleótidos, obteniéndose un alto contenido proteico con buen balance de aminoácidos, alta digestibilidad y mejor aprovechamiento de sus nutrientes. La inclusión de éstos hidrolizados en los alimentos acuícolas puede mejorar el crecimiento y la eficiencia alimentaria de los organismos acuáticos en cultivo. Este artículo presenta una revisión sobre investigaciones de la inclusión de hidrolizados proteicos de pescado en dietas experimentales para peces, crustáceos, moluscos y algas, y los efectos en los desempeños productivos, en los últimos veinte años.

Urban parks are green infrastructure elements that should contribute to improving the quality of life and well-being of citizens. In this work there are presented the results of applying a new index to estimate the potential allergenicity of parks located in 20 Spanish cities. This index, which considers intrinsic biological and biometric parameters of existing plant species in parks, allows the allergenic risk thereof to be calculated on a scale ranging from 0 to 1, depending on whether to the park’s allergenicity is zero or has a high risk for the population. The parks selected for this study have different typologies, sizes, species richness and biodiversities, which has yielded highly variable index values. Almost half of the analysed parks have an index value higher than 0.30, a threshold considered having a moderate to high risk, and therefore, enough to cause allergy symptoms in the population. Conversely, most of the parks had an index value below this threshold, so that the risk of suffering allergies is low or very low. The formula also allows the species that most contribute to the resulting value for allergenicity to be known, which are those having an anemophilous strategy of pollination, extended periods of flowering, and a referenced high allergenicity. These requirements are met by all species of the Betulaceae, Cupressaceae and Moraceae families, and to a lesser extent by Oleaceae and Platanaceae. It can be concluded that the development of an index to estimate the allergenicity of urban green spaces constitutes a useful tool to minimize the impact of pollen allergy on the population. Los parques urbanos son elementos de la infraestructura verde que deben contribuir a mejorar la calidad de vida y el bienestar ciudadano. En este trabajo se presentan los resultados de la aplicación de un novedoso índice que estima la alergenicidad potencial de las zonas verdes urbanas. Este índice, que contempla parámetros biológicos y biométricos intrínsecos a las especies arbóreas existentes en los parques, genera un resultado cuyo valor está comprendido entre 0 y 1 según el potencial alergénico del parque sea nulo o de riesgo alto para la población. En una primera fase el índice se ha aplicado a parques de diferente tipología, diseño, tamaño, riqueza específica y biodiversidad ubicados en 20 ciudades españolas. Los resultados han mostrado que algunos de los parques estudiados registran un valor de índice superior a 0.30, umbral suficiente para causar síntomas de alergia a la población expuesta, y por tanto, de riesgo moderado o alto. Por el contrario, en la mayoría de los parques se obtuvo un valor inferior a este umbral. También es posible conocer cuáles son las especies que más contribuyen al valor resultante, que son aquellas con estrategia de polinización anemófila, periodos de oración extensos y alta alergenicidad referenciada. Estos requisitos los cumplen todas las especies de las familias Betuláceas, Cupresáceas y Moráceas, y en menor extensión, Oleáceas y Platanáceas. Puede concluirse que el desarrollo de un índice de estimación de alergenicidad de espacios verdes urbanos constituye una herramienta de utilidad para minimizar el impacto de la alergia polínica en la población. Os parques urbanos são elementos da infraestrutura verde que devem contribuir para a melhoria da qualidade de vida dos cidadãos. Neste trabalho apresentam-se os resultados da aplicação de um índice inovador, que permite estimar a alergenicidade potencial dos espaços verdes urbanos. Este índice, que inclui parâmetros biológicos e biométricos, intrínsecos às próprias espécies existentes nos parques, tem como resultado um valor numa escala entre 0 e 1, de acordo com o potencial alergénico do parque, caso este seja, respetivamente e nos seus extremos, nulo ou de máximo risco para as populações. O índice foi aplicado em parques de diferentes tipologias, desenho, tamanho, riqueza específica e biodiversidade, situados em 20 cidades espanholas. Os resultados demonstram que alguns dos parques estudados registam um valor de índice superior a 0.30, limite su ciente para causar sintomas de alergia na população exposta, e por tanto, risco moderado a alto. No entanto, a maioria dos parques apresenta um valor inferior a este limite. Também é possível conhecer quais as espécies que mais contribuem para o valor do índice, que correspondem aquelas com a estratégia de polinização anemófila, períodos de oração extensos e potencial alergénico referenciado. Estes requisitos são aplicáveis a todas as espécies das famílias Betuláceas, Cupressáceas e Moráceas, e em menor medida, Oleáceas e Platanáceas. Pode assim concluir-se que a aplicação de um índice de previsão dos níveis de alergenicidade dos espaços verdes urbanos constitui uma ferramenta útil para minimizar o impacto da alergia polínica sobre a população.

The relationship was analyzed between Ca, Mg, Fe, Ni, Cu, Zn, Cr, Cd, and P and biochemical parameters (cholesterol, triglycerides, albumin, total protein, globulins, total thiols, ferritin, creatinine, glucose, urea, total bilirubin and the aspartate transaminase [AST] and alanine aminotransferase [ALT] enzymes) in vegetarians (25), regular consumers of seafood (RCSF, 30) and omnivores (30). The biochemical parameters were analyzed by using automated blood chemistry equipment, and the chemical elements were analyzed by Inductively Coupled Plasma (ICP) assay. No significant differences in Zn, Ca, and P concentrations were found between the groups. Biochemical parameters were observed to be within the reference ranges. Vegetarians showed the highest concentrations of Fe, Ca, Ni, Cr, Cu and Cd, the highest mean values of triglycerides and total thiols, and the lowest concentration of urea and ALT. In all groups, Cd and Fe were found to be significantly associated. Vegetarians showed an association with Mg, Zn, P, Ca and Ni. An association was found between P, Cr, Fe, Cd and Mg, and Zn, Cu and Ca in blood of the RCSF group, while omnivores presented an association with Mg, Cr, Ni, P, and Zn. Ferritin was associated with cadmium in RCSF, while in omnivores Cd and Ni were associated with AST and negatively associated with ferritin and Ca. Se analizó la relación entre Ca, Mg, Fe, Ni, Cu, Zn, Cr, Zn, Cd y P y los parámetros bioquímicos (colesterol, triglicéridos, albúmina, proteínas totales, globulinas, tioles totales, ferritina, creatinina, glicemia, urea, bilirrubina total y de las enzimas transaminasas aspartatoaminotransferasa (AST) y alaninaaminotransferasa (ALT) en ochenta y cinco (85) personas de ambos sexos (30 y 75 años), vegetarianos (25), consumidores habituales de productos del mar (CHPM, 30) y omnívoros (30). No se encontraron diferencias significativas en las concentraciones de Zn, Ca, y P entre los grupos. Los parámetros bioquímicos se observaron dentro de los rangos de referencia. Los vegetarianos mostraron las más altas concentraciones de Fe, Ca, Ni, Cr, Cu y Cd, los valores promedios más altos de triglicéridos y de tioles totales y los más bajos de urea y ALT. Se observó el Fe asociado al Cd en todos los grupos. Se observó asociación de Mg, Zn, P, Ca y Ni en vegetarianos, de P, Cr, Fe, Cd y Mg en los CHPM y de Mg, Cr, Ni, P, Zn en omnívoros. La ferritina estuvo asociada con el Cd en los CHPM. El Cd y el Ni se asocian con AST en omnívoros y negativamente con la ferritina y el Ca. Analisou-se a relação entre Ca, Mg, Fe, Ni, Cu, Zn, Cr , Cd, e P e os parâmetros bioquímicos [colesterol , triglicéridos , albumina , proteína total, globulinas , tióis totais, ferritina, creatinina, glicose, ureia, bilirrubina total e enzimas aspartato aminotransferase (AST) e alanina aminotransferase (ALT)] em 85 pessoas de ambos os sexos, com idades entres 30 e 75 anos; vegetarianos (25), consumidores regulares de produtos do mar (CRPM, 30) e omnívoros (30). Os parâmetros bioquímicos foram analisados usando equipamento automático de análise química do sangue e os elementos químicos foram analisados por ICP. Não foram encontradas diferenças significativas nas concentrações de Zn, Ca e P entre os grupos. Os parâmetros bioquímicos observados estavam dentro dos limites de referência. Os vegetarianos apresentaram as maiores concentrações de Fe, Ca , Ni , Cr , Cu e Cd , os valores médios mais elevados de triglicéridos e tióis totais e as concentrações mais baixas de ureia e ALT. Observou-se o Fe associado ao Cd em todos os grupos. Observou-se a associação entre Mg, Zn, P, Ca e Ni nos vegetarianos, entre P, Cr, Fe, Cd e Mg nos CRPM e entre Mg, Cr, Ni, P e Zn nos omnívoros. Verificou-se a associação da Ferritina com o Cd nos consumidores de produtos do mar. O Cd e o Ni associam-se às AST nos omnívoros e negativamente à ferritina e Ca.

Water temperature is a common variable of interest in stream ecology studies. In this paper, the performance of the ACR SmartButton thermometer and a low-cost protective case were evaluated for stream temperature measurement. The accuracy of the SmartButtons at 0, 10 and 15ºC was well within the ± 1ºC range specified by the manufacturer. For 50-60 % of the readings performed, the error was greater than the ± 0.2ºC correction factor accepted for meteorological temperature measurements. However, the observed level of accuracy is most likely sufficient for most biological applications if the loggers arecalibrated against a reference standard. Themetallic case that we usedhad a negligible effect on temperature measurements and offers a reliable way to protect the SmartButton during use in small streams. La temperatura del agua es una variable de interés en los estudios de ecología fluvial. En este trabajo,se evalúa el rendimiento de los termómetros ACR SmartButton y de una carcasa protectora de bajo coste para la medida de la temperatura en ríos. La precisión de los SmartButton a 0, 10 y 15ºC se encuentra dentro del rango de ± 1ºC que especifica el fabricante. El error observado en las lecturas sobrepasa el factor máximo de corrección de ± 0.2ºC que se acepta para la medida meteorológica de la temperatura en el 50-60 % de los datos recogidos. Sin embargo, la precisión observada es probablemente suficiente para otras aplicaciones biológicas, si los termómetros se calibran frente a una referencia estándar. Las carcasas protectoras que hemos utilizado tienen un efecto despreciable en las medidas de temperatura y representan un método seguro para instalar los SmartButtons en arroyos.

This investigation assesses the effects of a drought period on the physical and chemical properties and on the plankton community of a Mediterranean high mountain lake (Río Seco, Sierra Nevada, Spain). The study was performed over two consecutive years, 2005 and 2006. Both years were characterised by strong differences associated with rainfall patterns. The first year was extremely dry and had rainfall values far below the mean annual average of 813 mm that has been observed over the previous twenty years. The results of the study confirm that the dissolved inorganic nitrogen:soluble reactive phosphorus ratio (DIN:SRP) showed notable differences throughout both study years, with a greater nitrogen limitation during 2005. The ratio of total biovolume of autotrophic to heterotrophic organisms (A:H) also presented interannual differences. The A:H ratio was generally below 1 during 2005, whereas it was above this value in 2006. Therefore, the main plankton component was mesozooplankton in 2005 and phytoplankton in 2006. Ciliates and bacteria showed a mean biovolume significantly higher in 2005, whereas the biovolume of heterotrophic nanoflagellates was similar during both years. These results indicate a prevalent "top-down" control by zooplankton during the dry year and, on the contrary, a predominant "bottom-up" control during the wetter year as a consequence of the nutrient limitation of primary production. En este trabajo se evalúan los efectos de un periodo de sequía sobre las propiedades físicas y químicas y las comunidades planctónicas de una laguna mediterránea de alta montaña (Río Seco, Sierra Nevada, España). El estudio se extendió durante los meses estivales de 2005 y 2006. Ambos años mostraron marcadas diferencias en cuanto al nivel de precipitaciones, estando el primero caracterizado por una pronunciada sequía, con valores de precipitación muy inferiores a los valores medios anuales de 813 mm registrados en los últimos 20 años. Los resultados obtenidos mostraron cómo la disponibilidad de nutrientes en el agua, expresada a través de la razón entre nitrógeno inorgánico disuelto y fósforo soluble reactivo (DIN:SRP), experimentó notables diferencias entre los dos años de estudio, mostrando una limitación por nitrógeno más acentuada durante el verano de 2005. La razón entre biovolumen total de organismos autótrofos y heterótrofos (A:H) también mostró diferencias entre ambos años. Esta razón fue generalmente inferior a uno en 2005 y superior a este valor en 2006, siendo el mesozooplancton el componente principal del plancton en 2005 y el fitoplancton en 2006. Ciliados y bacterias mostraron un biovolumen medio significativamente mayor en 2005, mientras que el biovolumen de heterótrofos nanoflagelados fue prácticamente constante en ambos años. Estos resultados, indican un predominio del control de tipo "top-down" del zooplancton en el año más seco, mientras que el control fue predominantemente de tipo "bottom-up" en el año más húmedo, a consecuencia de una mayor limitación de nutrientes.

Despite the Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway being frequently altered in T-ALL/LBL, no specific therapy has been approved for T-ALL/LBL patients with constitutive signalling by JAK/STAT, so there is an urgent need to identify pathway members that may be potential therapeutic targets. In the present study, we searched for JAK/STAT pathway members potentially modulated through aberrant methylation and identified SOCS3 hypermethylation as a recurrent event in T-ALL/LBL. Additionally, we explored the implications of SOCS3 deregulation in T-ALL/LBL and demonstrated that SOCS3 counteracts the constitutive activation of the JAK/STAT pathway through different molecular mechanisms. Therefore, SOCS3 emerges as a potential therapeutic target in T-ALL/LBL, Comunidad de Madrid, Grant/Award Number: B2017/BMD-3778; LINFOMAS-CM; Fundación Científica Asociación Española Contra el Cáncer, Grant/Award Number: PROYE18054PIRI; Fundación Ramón Areces, Grant/Award Number: CIVP19S7917; Instituto de Investigación Sanitaria Fundación Jiménez Díaz; Ministerio de Ciencia, Innovación y Universidades, Grant/ Award Number: RTI2018- 093330-B-I00 and MCIU/FEDER; Ministerio de Economía y Competitividad, Grant/Award Number: SAF2015-70561-R and MINECO/FEDER

The emergent human coronavirus SARS-CoV-2 and its high infectivity rate has highlighted the strong need for new virucidal treatments. In this sense, the use of photodynamic therapy (PDT) with white light, to take advantage of the sunlight, is a potent strategy for decreasing the virulence and pathogenicity of the virus. Here, we report the virucidal effect of PDT based on Hypericum extract (HE) in combination with white light, which exhibits an inhibitory activity of the human coronavirus HCoV-229E on hepatocarcinoma Huh-7 cells. Moreover, despite continuous exposure to white light, HE has long durability, being able to maintain the prevention of viral infection. Given its potent in vitro virucidal capacity, we propose HE in combination with white light as a promising candidate to fight against SARS-CoV-2 as a virucidal compound., This research was funded by Fundación Universidad Autónoma de Madrid, grant number PI21/00315 and by Instituto de Salud Carlos iii, grant number PI21/00953.

A striking feature of the nervous system pertains to the appearance of different neural cell subtypes at different axial levels. Studies in the Drosophila central nervous system reveal that one mechanism underlying such segmental differences pertains to the segment-specific removal of cells by programmed cell death (PCD). One group of genes involved in segment-specific PCD is the Hox homeotic genes. However, while segment-specific PCD is highly precise, Hox gene expression is evident in gradients, raising the issue of how the Hox gene function is precisely gated to trigger PCD in specific segments at the outer limits of Hox expression. The Drosophila Va neurons are initially generated in all nerve cord segments but removed by PCD in posterior segments. Va PCD is triggered by the posteriorly expressed Hox gene Abdominal-B (Abd-B). However, Va PCD is highly reproducible despite exceedingly weak Abd-B expression in the anterior frontiers of its expression. Here, we found that the transcriptional cofactor Dachshund supports Abd-B-mediated PCD in its anterior domain. In vivo bimolecular fluorescence complementation analysis lends support to the idea that the Dachshund/Abd-B interplay may involve physical interactions. These findings provide an example of how combinatorial codes of transcription factors ensure precision in Hox-mediated PCD in specific segments at the outer limits of Hox expression, Ministerio de Ciencia y Educación, Grant/Award Number: PID2019-110952GB-I00

Regeneration, the ability to replace lost body parts, is a widespread phenomenon in the animal kingdom often connected to asexual reproduction or fission, since the only difference between the two appears to be the stimulus that triggers them. Both developmental processes have largely been characterized; however, the molecular toolkit and genetic mechanisms underlying these events remain poorly unexplored. Annelids, in particular the oligochaete Pristina leidyi, provide a good model system to investigate these processes as they show diverse ways to regenerate, and can reproduce asexually through fission under laboratory conditions. Here, we used a comparative transcriptomics approach based on RNA‐sequencing and differential gene expression analyses to understand the molecular mechanisms involved in anterior regeneration and asexual reproduction. We found 291 genes upregulated during anterior regeneration, including several regeneration‐related genes previously reported in other annelids such as frizzled, paics, and vdra. On the other hand, during asexual reproduction, 130 genes were found upregulated, and unexpectedly, many of them were related to germline development during sexual reproduction. We also found important differences between anterior regeneration and asexual reproduc tion, with the latter showing a gene expression profile more similar to that of control individuals. Nevertheless, we identified 35 genes that were upregulated in both conditions, many of them related to cell pluripotency, stem cells, and cell proliferation. Overall, our results shed light on the molecular mechanisms that control anterior regeneration and asexual reproduction in annelids and reveal similarities with other animals, suggesting that the genetic machinery controlling these processes is conserved across metazoans., Irene del Olmo was supported by the program “Ayudas para el Fomento de la Investigación en Estudios de Máster‐UAM 2020 en la Facultad de Ciencias” financed by the Universidad Autónoma de Madrid (3931‐ 326F‐726DP6A38‐4F77). Aida Verdes was funded by the European Union's Horizon 2020 research and innovation program through a Marie Skłodowska‐Curie individual fellowship (grant 841576). This study was funded through the Madrid Government (Comunidad de Madrid‐Spain) under the Multiannual Agreement with Universidad Autónoma de Madrid in the line of action encouraging youth research doctors, in the context of the V PRICIT (Regional Program of Research and Technological Innovation) (SI1/PJI/2019‐00532), and the EMBO Long Term Fellowship (ALTF‐217‐2018) to Patricia Álvarez‐Campos.

Propionic acidemia (PA) disorder shows major involvement of the heart, among other alterations. A significant number of PA patients develop cardiac complications, and available evidence suggests that this cardiac dysfunction is driven mainly by the accumulation of toxic metabolites. To contribute to the elucidation of the mechanistic basis underlying this dysfunction, we have successfully generated cardiomyocytes through the differentiation of induced pluripotent stem cells (iPSCs) from a PCCB patient and its isogenic control. In this human cellular model, we aimed to examine microRNAs (miRNAs) profiles and analyze several cellular pathways to determine miRNAs activity patterns associated with PA cardiac phenotypes. We have identified a series of upregulated cardiac-enriched miRNAs and alterations in some of their regulated signaling pathways, including an increase in the expression of cardiac damage markers and cardiac channels, an increase in oxidative stress, a decrease in mitochondrial respiration and autophagy; and lipid accumulation. Our findings indicate that miRNA activity patterns from PA iPSC-derived cardiomyocytes are biologically informative and advance the understanding of the molecular mechanisms of this rare disease, providing a basis for identifying new therapeutic targets for intervention strategies.

The synthetic compound fasentin has been described as a modulator of GLUT-1 and GLUT-4 transporters, thus inhibiting glucose uptake in some cancer cells. Endothelial glucose metabolism has been recently connected to angiogenesis and it is now an emerging topic in scientific research. Indeed, certain compounds with a known effect on glucose metabolism have also been shown to inhibit angiogenesis. In this work we tested the capability of fasentin to modulate angiogenesis in vitro and in vivo. We show that fasentin inhibited tube formation in endothelial cells by a mechanism that involves a negative effect on endothelial cell proliferation and invasion, without affecting other steps related to the angiogenic process. However, fasentin barely decreased glucose uptake in human dermal microvascular endothelial cells and the GLUT-1 inhibitor STF-31 failed to inhibit tube formation in these cells. Therefore, this modulatory capacity on endothelial cells function exerted by fasentin is most likely independent of a modulation of glucose metabolism. Taken together, our results show a novel biological activity of fasentin, which could be evaluated for its utility in cancer and other angiogenesis-dependent diseases.

In eukaryotic cells, a subset of cell surface proteins is attached by the glycolipid glycosylphosphatidylinositol (GPI) to the external leaflet of the plasma membrane where they play important roles as enzymes, receptors, or adhesion molecules. Here we present a protocol for purification and mass spectrometry analysis of the lipid moiety of individual GPI-anchored proteins (GPI-APs) in yeast. The method involves the expression of a specific GPI-AP tagged with GFP, solubilization, immunoprecipitation, separation by electrophoresis, blotting onto PVDF, release and extraction of the GPI-lipid moiety and analysis by mass spectrometry. By using this protocol, we could determine the precise GPI-lipid structure of the GPI-AP Gas1-GFP in a modified yeast strain. This protocol can be used to identify the lipid composition of the GPI anchor of distinct GPI-APs from yeast to mammals and can be adapted to determine other types of protein lipidation., This research was funded by the FEDER/Ministerio de Ciencia, Innovación y Universidades—Agencia Estatal de Investigación/BFU2017-89700-P to Manuel Muñiz, "VI Own Research Plan" of the University of Seville VIPPIT-2020-I.5 to Manuel Muñiz, by the Incentivo al Grupo de Investigación BIO-271 (2019/BIO-271) and the NCCR Chemical Biology and the Swiss National Science Foundation (51NF40-185898 and 310030_184949) to Howard Riezman. ‘V Own Research Plan’ of the University of Seville (VPPI-US) contract (cofounded by the European Social Fund) to Sergio Lopez, University of Seville fellowships to Sofia Rodriguez-Gallardo and Ana Maria Perez-Linero, Ministry of Education, Culture and Sport (MECD) fellowship to Susana Sabido-Bozo and contract from the University of Seville by the Youth Employment Initiative to Alejandro Cortes-Gomez.

Motivation: We accessed published and unpublished floristic sources to compile a comprehensive species list of the Iberian–Balearic terrestrial vascular flora and generate AFLIBER, an accurate floristic database of georeferenced plant occurrence records. Main type of variable contained: Species distribution data totalling 1,824,549 plant occurrence records corresponding to 6,456 species and subspecies. Spatial location and grain: The western Mediterranean, including inland territories of Spain, Portugal and Andorra and the adjacent archipelagos of Berlengas, Columbretes and the Balearic Islands, covered by 6,316 UTM quadrangular grid cells of 10 km resolution. Time period: All distributional trustable records were considered to create the AFLIBER database, most of them dating from the 1960s onwards. Major taxa and level of measurement: Terrestrial vascular plant species and subspecies. Software format: Data are supplied as comma-separated text (csv) files, Ministerio de Economía, Industria y Competitividad, Gobierno de España, Grant/ Award Number: CGL2017-86926-P

Carex section Schoenoxiphium (Cariceae, Cyperaceae) is endemic to the Afrotropical biogeographic region and is mainly distributed in southern and eastern Africa, with its center of diversity in eastern South Africa. The taxon was formerly recognized as a distinct genus and has a long history of taxonomic controversy. It has also an important morphological and molecular background in particular dealing with the complexity of its inflorescence and the phylogenetic relationships of its species. We here present a fully updated and integrative monograph of Carex section Schoenoxiphium based on morphological, molecular and cytogenetic data. A total of 1,017 herbarium specimens were examined and the majority of the species were studied in the field. Previous molecular phylogenies based on Sanger-sequencing of four nuclear and plastid DNA regions and RAD-seq were expanded. For the first time, chromosome numbers were obtained, with cytogenetic counts on 44 populations from 15 species and one hybrid. Our taxonomic treatment recognizes 21 species, one of them herein newly described (C. gordon-grayae). Our results agree with previous molecular works that have found five main lineages in Schoenoxiphium. We provide detailed morphological descriptions, distribution maps and analytical drawings of all accepted species in section Schoenoxiphium, an identification key, and a thorough nomenclatural survey including 19 new typifications and one nomen novum.

Glutaminase (GA) catalyzes the first step in mitochondrial glutaminolysis playing a key role in cancer metabolic reprogramming. Humans express two types of GA isoforms: GLS and GLS2. GLS isozymes have been consistently related to cell proliferation, but the role of GLS2 in cancer remains poorly understood. GLS2 is repressed in many tumor cells and a better understanding of its function in tumorigenesis may further the development of new therapeutic approaches. We analyzed GLS2 expression in HCC, GBM and neuroblastoma cells, as well as in monkey COS-7 cells. We studied GLS2 expression after induction of differentiation with phorbol ester (PMA) and transduction with the full-length cDNA of GLS2. In parallel, we investigated cell cycle progression and levels of p53, p21 and c-Myc proteins. Using the baculovirus system, human GLS2 protein was overexpressed, purified and analyzed for posttranslational modifications employing a proteomics LC-MS/MS platform. We have demonstrated a dual targeting of GLS2 in human cancer cells. Immunocytochemistry and subcellular fractionation gave consistent results demonstrating nuclear and mitochondrial locations, with the latter being predominant. Nuclear targeting was confirmed in cancer cells overexpressing c-Myc- and GFP-tagged GLS2 proteins. We assessed the subnuclear location finding a widespread distribution of GLS2 in the nucleoplasm without clear overlapping with specific nuclear substructures. GLS2 expression and nuclear accrual notably increased by treatment of SH-SY5Y cells with PMA and it correlated with cell cycle arrest at G2/M, upregulation of tumor suppressor p53 and p21 protein. A similar response was obtained by overexpression of GLS2 in T98G glioma cells, including downregulation of oncogene c-Myc. Furthermore, human GLS2 was identified as being hypusinated by MS analysis, a posttranslational modification which may be relevant for its nuclear targeting and/or function. Our studies provide evidence for a tumor suppressor role of GLS2 in certain types of cancer. The data imply that GLS2 can be regarded as a highly mobile and multilocalizing protein translocated to both mitochondria and nuclei. Upregulation of GLS2 in cancer cells induced an antiproliferative response with cell cycle arrest at the G2/M phase.

© 2020 by the authors., Photodynamic Therapy (PDT) with methyl-aminolevulinate (MAL-PDT) is being used for the treatment of Basal Cell Carcinoma (BCC), although resistant cells may appear. Normal differentiated cells depend primarily on mitochondrial oxidative phosphorylation (OXPHOS) to generate energy, but cancer cells switch this metabolism to aerobic glycolysis (Warburg effect), influencing the response to therapies. We have analyzed the expression of metabolic markers (β-F1-ATPase/GAPDH (glyceraldehyde-3-phosphate dehydrogenase) ratio, pyruvate kinase M2 (PKM2), oxygen consume ratio, and lactate extracellular production) in the resistance to PDT of mouse BCC cell lines (named ASZ and CSZ, heterozygous for ptch1). We have also evaluated the ability of metformin (Metf), an antidiabetic type II compound that acts through inhibition of the AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway to sensitize resistant cells to PDT. The results obtained indicated that resistant cells showed an aerobic glycolysis metabolism. The treatment with Metf induced arrest in the G0/G1 phase and a reduction in the lactate extracellular production in all cell lines. The addition of Metf to MAL-PDT improved the cytotoxic effect on parental and resistant cells, which was not dependent on the PS protoporphyrin IX (PpIX) production. After Metf + MAL-PDT treatment, activation of pAMPK was detected, suppressing the mTOR pathway in most of the cells. Enhanced PDT-response with Metf was also observed in ASZ tumors. In conclusion, Metf increased the response to MAL-PDT in murine BCC cells resistant to PDT with aerobic glycolysis., This research was funded by Spanish grants from Instituto de Salud Carlos III MINECO and Feder Funds (FIS PI15/00974 and PI18/00708) and Ministerio de Ciencia, Innovación y Universidades (SAF2016-75916-R).

Background: Identification of signaling pathways altered at early stages after cardiac ischemia/reperfusion (I/R) is crucial to develop timely therapies aimed at reducing I/R injury. The expression of G protein-coupled receptor kinase 2 (GRK2), a key signaling hub, is up-regulated in the long-term in patients and in experimental models of heart failure. However, whether GRK2 levels change at early time points following myocardial I/R and its functional impact during this period remain to be established. Methods: We have investigated the temporal changes of GRK2 expression and their potential relationships with the cardioprotective AKT pathway in isolated rat hearts and porcine preclinical models of I/R. Findings: Contrary to the maladaptive up-regulation of GRK2 reported at later times after myocardial infarction, successive GRK2 phosphorylation at specific sites during ischemia and early reperfusion elicits GRK2 degradation by the proteasome and calpains, respectively, thus keeping GRK2 levels low during early I/R in rat hearts. Concurrently, I/R promotes decay of the prolyl-isomerase Pin1, a positive regulator of AKT stability, and a marked loss of total AKT protein, resulting in an overall decreased activity of this pro-survival pathway. A similar pattern of concomitant down-modulation of GRK2/AKT/Pin1 protein levels in early I/R was observed in pig hearts. Calpain and proteasome inhibition prevents GRK2/Pin1/AKT degradation, restores bulk AKT pathway activity and attenuates myocardial I/R injury in isolated rat hearts. Interpretation: Preventing transient degradation of GRK2 and AKT during early I/R might improve the potential of endogenous cardioprotection mechanisms and of conditioning strategies., Instituto de Salud Carlos III, Spain (grant PI-16/00232; RETICS-RIC-RD12/0042/0021 to DGD, cofunded with European Regional Development Fund-FEDER contribution, and grants PI14-00435 and PI17-00576 to PP), by Ministerio de Economía; Industria y Competitividad (MINECO) of Spain (grant SAF2017-84125-R to F.M.); by CIBERCV-Instituto de Salud Carlos III, Spain (grant CB16/11/00479 to DGD and CB16/11/00278 to F.M, cofunded with European Regional Development Fund-FEDER contribution), and Programa de Actividades en Biomedicina de la Comunidad de Madrid-B2017/BMD-3671-INFLAMUNE to F.M. We also acknowledge institutional support to the CBMSO from Fundación Ramón Areces

[Background] Viruses are key players regulating microbial ecosystems. Exploration of viral assemblages is now possible thanks to the development of metagenomics, the most powerful tool available for studying viral ecology and discovering new viruses. Unfortunately, several sources of bias lead to the misrepresentation of certain viruses within metagenomics workflows, hindering the shift from merely descriptive studies towards quantitative comparisons of communities. Therefore, benchmark studies on virus enrichment and random amplification protocols are required to better understand the sources of bias. [Results] We assessed the bias introduced by viral enrichment on mock assemblages composed of seven DNA viruses, and the bias from random amplification methods on human saliva DNA viromes, using qPCR and deep sequencing, respectively. While iodixanol cushions and 0.45 μm filtration preserved the original composition of nuclease-protected viral genomes, low-force centrifugation and 0.22 μm filtration removed large viruses. Comparison of unamplified and randomly amplified saliva viromes revealed that multiple displacement amplification (MDA) induced stochastic bias from picograms of DNA template. However, the type of bias shifted to systematic using 1 ng, with only a marginal influence by amplification time. Systematic bias consisted of over-amplification of small circular genomes, and under-amplification of those with extreme GC content, a negative bias that was shared with the PCR-based sequence-independent, single-primer amplification (SISPA) method. MDA based on random priming provided by a DNA primase activity slightly outperformed those based on random hexamers and SISPA, which may reflect differences in ability to handle sequences with extreme GC content. SISPA viromes showed uneven coverage profiles, with high coverage peaks in regions with low linguistic sequence complexity. Despite misrepresentation of certain viruses after random amplification, ordination plots based on dissimilarities among contig profiles showed perfect overlapping of related amplified and unamplified saliva viromes and strong separation from unrelated saliva viromes. This result suggests that random amplification bias has a minor impact on beta diversity studies. [Conclusions] Benchmark analyses of mock and natural communities of viruses improve understanding and mitigate bias in metagenomics surveys. Bias induced by random amplification methods has only a minor impact on beta diversity studies of human saliva viromes., Institutional grants from the Fundación Ramón Areces and Banco de Santander to the CBMSO are acknowledged.This work was supported by the Spanish Ministry of Economy and Competitiveness through grant SAF2012-38421 and a “Formación de Personal Investigador” Ph.D. studentship to MP-M

Root-knot nematodes (RKNs) are obligate endoparasites that feed on their host plants to complete its life cycle, representing a major threat to agriculture and economy worldwide. The development of new management strategies becomes essential as effective chemical nematicides are progressively being restricted. Hence, we analysed grape pomace-derived biochars, pyrolysed at 350 °C (BC350) and 700 °C (BC700), focusing on their potential for RKN control. The thermal treatment of grape pomace caused an increase in the concentration of carbon and plant macro- and micronutrients, which were largely present in a water-soluble form. Synchrotron radiation-based Fourier transform infrared microspectroscopy data showed a general loss of carboxylic functional groups during pyrolysis, partially contributing to the alkalinisation of both biochars, mostly in BC700. Scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy analysis revealed a highly porous structure filled with different crystals composed of elements such as K, Ca, Mg, P, Si or Al, which could be a suitable environment for the growth of microorganisms. Biochar-derived aqueous extracts showed phytotoxicity to tomato seedlings at high concentrations, and disappeared upon dilution, but no toxic effect was observed on the nematode’s infective stage. However, the infective and reproductive traits of a Meloidogyne javanica population in tomato were significantly reduced (i.e. egg masses and eggs per plant) in washed-biochar-treated soil in pots (0.75%; BC350W). Therefore, the large amount of grape waste generated after wine production can be transformed into a valuable product such as biochar, effective for RKNs control, thus reducing the waste management problem and contributing to a circular economy, This work was supported by the Spanish Government (PID2019-105924RB-I00 MCIN/AEI/10.13039/501100011033; RED2018-102407-T) to CE, and (AGL201453771-R; AGL2017-87591-R) to LEH, and by the Castilla-La Mancha Govern ment (SBPLY/17/180501/000287 and SBPLY/21/180501/000033) to CE. AM-G was recipient of a PhD grant from Fundación Tatiana Pérez de Guzmán el Bueno. EU FEDER funds complemented all the grants

Health is a multi-dimensional concept, capturing how people feel and function. The broad concept of Active and Healthy Ageing was proposed by the World Health Organisation (WHO) as the process of optimizing opportunities for health to enhance quality of life as people age. It applies to both individuals and population groups. A universal Active and Healthy Ageing definition is not available and it may differ depending on the purpose of the definition and/or the questions raised. While the European Innovation Partnership on Active and Healthy Ageing (EIP on AHA) has had a major impact, a definition of Active and Healthy Ageing is urgently needed. A meeting was organised in Montpellier, France, October 20-21, 2014 as the annual conference of the EIP on AHA Reference Site MACVIA-LR (Contre les Maladies Chroniques pour un Vieillissement Actif en Languedoc Roussillon) to propose an operational definition of Active and Healthy Ageing including tools that may be used for this. The current paper describes the rationale and the process by which the aims of the meeting will be reached. Health is a multi-dimensional concept, capturing how people feel and function. The broad concept of Active and Healthy Ageing was proposed by the World Health Organisation (WHO) as the process of optimizing opportunities for health to enhance quality of life as people age. It applies to both individuals and population groups. A universal Active and Healthy Ageing definition is not available and it may differ depending on the purpose of the definition and/or the questions raised. While the European Innovation Partnership on Active and Healthy Ageing (EIP on AHA) has had a major impact, a definition of Active and Healthy Ageing is urgently needed. A meeting was organised in Montpellier, France, October 20-21, 2014 as the annual conference of the EIP on AHA Reference Site MACVIA-LR (Contre les Maladies Chroniques pour un Vieillissement Actif en Languedoc Roussillon) to propose an operational definition of Active and Healthy Ageing including tools that may be used for this. The current paper describes the rationale and the process by which the aims of the meeting will be reached.

Photodynamic Therapy (PDT) with methyl-aminolevulinate acid (MAL-PDT) is being used for the treatment of Basal cell carcinoma (BCC), but recurrences have been reported. In this work, we have evaluated resistance mechanisms to MAL-PDT developed by three BCC cell lines (ASZ, BSZ and CSZ), derived from mice on a ptch+/− background and with or without p53 expression, subjected to 10 cycles of PDT (10thG). The resistant populations showed mesenchymal-like structure and diminished proliferative capacity and size compared to the parental (P) cells. The resistance was dependent on the production of the endogenous photosensitiser protoporphyrin IX in the CSZ cell line and on its cellular localisation in ASZ and BSZ cells. Moreover, resistant cells expressing the p53 gene presented lower proliferation rate and increased expression levels of N-cadherin and Gsk3β (a component of the Wnt/β-catenin pathway) than P cells. In contrast, 10thG cells lacking the p53 gene showed lower levels of expression of Gsk3β in the cytoplasm and of E-cadherin and β-catenin in the membrane. In addition, resistant cells presented higher tumorigenic ability in immunosuppressed mice. Altogether, these results shed light on resistance mechanisms of BCC to PDT and may help to improve the use of this therapeutic approach.

Objetivos. 1. Caracterización de la longitud telomérica y de la expresión y actividad de la telomerasa en el pez cebra (Danio rerio) a lo largo de su ciclo de vida. 2. Caracterización de la línea de pez cebra deficiente en la subunidad catalítica de la telomerasa (tert). 3. Estudio de la longitud telomérica y de la expresión y actividad de la telomerasa en el pez cebra (Danio rerio) durante el proceso de regeneración de la aleta caudal del pez cebra salvaje y del pez cebra tert-deficiente. - Metodología. Para el primer objetivo, se han utilizado peces cebra de tipo salvaje, de diferentes estirpes y edades. El estudio de la expresión de genes se ha llevado a cabo mediante RT-PCR y PCR a tiempo real, el estudio de la actividad telomerasa mediante una técnica específica y cuantitativa llamada Q-TRAP basada en la amplificación de la secuencia telomérica. La longitud telomérica se ha cuantificado por técnicas diferentes: TRF que consiste en una hibridación con una sonda telomérica radioactiva, Q-FISH y Flow-FISH que consisten en una hibridación in situ con una sonda telomérica fluorescente. Para el cumplimiento del segundo objetivo, se han utilizado los genotipos, tert+/+, tert+/- y tert-/-. Además de las técnicas antes citadas, se han realizado curvas de supervivencia y un estudio histológico sobre diferentes tejidos (Hematoxilina/Eosina, PAS y TUNEL). El estudio de la G2 mutante se ha llevado a cabo con curvas de supervivencia, ensayo de apoptosis por TUNEL, y Q-FISH en núcleos metafásicos. Para el tercer objetivo, con los mismos genotipos y a diferentes edades. Se ha realizado un ensayo de detección de los círculos de ADN telomérico, un ensayo de regeneración en adultos con inhibición de un gen utilizando un morfolino seguido de electroporación o en larvas con inhibición de un gen por inmersión. Peces cebra han sido mantenidos según se indica en el manual del pez cebra. Todos los experimentos cumplen con la directiva de la Unión Europea (86/609/EU) y han sido aprobados por el Comité de Bioética del Hospital Clínico Universitario “Virgen de la Arrixaca” (España) y por el Comité Institucional del Cuidado y Uso Animal del Hospital Infantil de Boston (EEUU). - Resultados y conclusiones. Los resultados de esta tesis han demostrado la estrecha relación de la expresión de la telomerasa y la longitud de los telómeros durante la vida del pez, concluyendo que estos dos parámetros pueden ser utilizados como biomarcadores de envejecimiento en el pez cebra. La línea de pez cebra deficiente en telomerasa ha mostrado signos de envejecimiento prematuro, como curvatura de la columna, infertilidad y reducción de la vida media de la G1, apoyando al pez cebra como un modelo prometedor para estudiar el envejecimiento inducido por el telómero. Además, la G1 muestra activación de p53 en respuesta al desgaste telomerico, reproduce el fenómeno de anticipación genética, ligado al acortamiento telomérico observado en humanos con DC, debido a haploinsuficiencia de la telomerasa. La G2 muere en las primeras etapas de desarrollo y la restauración de la actividad telomerasa rescata la longitud de los telómeros y la supervivencia, indicando que la dosis de telomerasa es crucial. La inhibición genética de p53 rescata los efectos adversos de la perdidad de telómeros, indicando que los mecanismos moleculares están conservados desde peces hasta mamíferos. El estudio de regeneración demuestra que los genotipos tert+/+ y tert-/- regeneran después de una o varias escisiones, aunque la eficiencia de regeneración disminuye con el envejecimiento en ambos. La línea tert-/- regenera con un ratio de regeneración menor que tert+/+, y mantiene la longitud telomérica. ALT está implicado en el mantenimiento de los telómeros durante la regeneración de la aleta caudal en el pez mutante, y se necesitan más estudios para determinar el papel de la telomerasa y posiblemente de ALT en el pez cebra salvaje durante este proceso., Objectives. The specific objectives of the present work are: 1. Characterization of telomere length, telomerase expression and activity in zebrafish (Danio rerio) throughout its life cycle. 2. Characterization of the tert-deficient zebrafish line. 3. Study of telomere length, telomerase expression and activity in zebrafish (Danio rerio) during the caudal fin regeneration process in wild-type and tert mutant - Methodology. For approaching the first goal, wild-type zebrafish from different strains and different ages have been used. The study of gene expression was carried out by RT-PCR and real time PCR. The telomerase activity was determined by using a specific and quantitative assay based in the amplification of the telomere sequence called Q-TRAP, and also analyzed quantitatively by TRAP. The telomere length was quantified using three different techniques: by a whole-mount in situ hybridization with a fluorescent telomeric probe (Q-FISH and Flow-FISH) or by hybridization using a radioactive telomeric probe (TRF assay). To address the second goal, three zebrafish genotypes: tert+/+, tert+/- y tert-/- have been used. Besides the above techniques, survival curves to estimate the mean lifespan of the first generation (G1) of tert mutant line, and a histological study of different tissues (Hematoxylin / Eosin, PAS and TUNEL staining) have been performed. To study the tert mutant line G2: survival curves, apoptosis by TUNEL assay, and Q-FISH on metaphasic nucleus. To achieve the third objective, we have used all three genotypes at different ages. Telomere length was analyzed by Flow-FISH, and the detection of telomeric DNA circles by CCassay. A caudal fin regeneration assay on adult with gene inhibition using morpholino following by electroporation was performed and finally a caudal fin regeneration assay on larvae with gene inhibition by immersion. Zebrafish were maintained as described in the zebrafish handbook. The experiments performed comply with the Guidelines of the European Union Council (86/609/EU). Experiments and procedures were performed as approved by the Bioethical Committee of the University Hospital “Virgen de la Arrixaca” (Spain) and by the Children's Hospital Boston institutional Animal Care and Use Committee (USA). - Results and conclusions. The results of this tesis have shown that the expression of telomerase and telomere length are closely related during the entire life cycle of the fish and that these two parameters can be used as biomarkers of aging in zebrafish. Telomerase-deficient zebrafish characterization have shown premature aging and reduced lifespan in the first generation, as occurs in humans and may be considered as a promising model to study telomere-driven aging. Interestingly, this mutant line reproduced the genetic anticipation phenomenon, related to telomere shortening, observed in humans with DC. The second-generation embryos died in early developmental stages, and restoration of telomerase activity rescued telomere length and survival, indicating that telomerase dosage is crucial. Genetic inhibition of p53 rescued the adverse effects of telomere loss, indicating that the molecular mechanisms induced by telomere shortening are conserved from fish to mammals. The regeneration study showed that all genotypes regenerated after single or successive amputations and that regeneration efficiency decreased with aging. Tert mutant line showed a lower regeneration rate than its wild-type counterpart, and maintained telomere length in the regenerative tissue. ALT mechanism is implicated in telomere maintenance during caudal fin regeneration in telomerase-deficient fish. Further studies are necessary to definitively elucidate the role of telomerase and the possible ALT implication in telomere maintenance during caudal fin regeneration in wild-type animals.

Un área muy actual de investigación es el estudio de las causas del envejecimiento. En el ámbito de la biología reproductiva también lo está siendo por sus implicaciones tanto en la medicina como en la veterinaria. Si durante muchos años los estudios se han centrado en la mujer o en las hembras para explicar el descenso reproductivo con la edad actualmente se está prestando más atención al factor masculino. Estos estudios abordan varios aspectos: cambios endocrinos, alteraciones de los espermatozoides, trasmisión de enfermedades hereditarias, o la necesidad de terapias sustitutivas de testosterona en la vejez. Respecto a los cambios histológicos del testículo envejecido, se han realizado diversos estudios desde hace décadas, pero han dejado ciertas cuestiones sin aclarar y se han centrado en el hombre y en la rata. Junto a esto, durante los últimos años, diversos autores han propuesto que el mecanismo tisular implicado en la progresiva disminución de la espermatogénesis con la edad, está relacionado con cambios en el equilibrio entre la proliferación/apoptosis en el epitelio seminífero. En la presente tesis doctoral nos hemos propuesto por un lado, realizar un completo análisis histológico tanto cualitativo como cuantitativo de los cambios que acontecen con la edad en el epitelio seminífero y pared tubular del hámster sirio. Por otro, hemos investigado también si la proliferación está o no alterada en este epitelio relacionándola con las alteraciones histológicas observadas en el mismo. Para ello, utilizamos hámsteres machos de 6, 12 y 24 meses de edad. Los testículos fueron fijados y procesados para microscopia de luz convencional e inmunohistoquimica y, electrónica de transmisión. Se realizó un estudio semicuantitativo del tipo histológico de degeneración de las secciones de los túbulos seminíferos determinando el grado de progresiva alteración de su epitelio con la edad. Inmunohistoquímicamente se identificó la inmunoreactividad a laminina y fibronectina en la pared tubular y las células en proliferación. Morfométricamente se calculó una serie de parámetros testiculares y el volumen total de inmunoreactividad a fibronectina y laminina. Respecto a la proliferación se calculó el índice total de proliferación como el número de células proliferando por tipo histológico de sección tubular. Por último, con las secciones ultrafinas se realizó una descripción ultrarestructural del epitelio seminífero y se calculó el grosor de la pared tubular. En los animales envejecidos se observó un incremento significativo de las secciones tubulares con diversas alteraciones del epitelio seminífero especialmente hipoespermatogénesis y detención de la espermatogénesis. El volumen del testículo, el diámetro y volumen del túbulo como el volumen de epitelio del túbulo seminífero sufrieron un descenso significativo con la edad, por el contario la longitud del túbulo se incrementó. El volumen de fibronectina y laminina aumento en los animales viejos siendo el índice de proliferación menor en estos animales. El número de células en proliferación por tipo histológico de sección de túbulo seminífero fue progresivamente y significativamente menor en las secciones alteradas respecto a las secciones normales. No se observó diferencias significativas respecto a la edad entre tipos histológicos de sección de túbulo seminífero. Ultraestructuralmente se observó alteraciones citológicas importantes en la célula de Sertoli así como en el proceso de espermiogénesis. La pared presentó un engrosamiento significativo tanto en los túbulos normales como en los hipoespermatogénicos o en detención de la espermatogénesis. En conclusión, el envejecimiento del túbulo seminífero del hámster supone una pérdida progresiva de la actividad espermatogénica que está relacionada con cambios del túbulo y de su pared. Estos cambios no se producen de forma uniforme en todo él y se asocian a una disminución de la proliferación celular en sus zonas alteradas histológicamente. Éstas presentan además células de Sertoli modificadas ultraestructuralmente., A very current area of research is the study of the causes of aging. In the field of reproductive biology the aging is important by their implications both in medicine and veterinary. If for many years the studies have focused on women or females to explain the reproductive decline with age it is now being paid more attention to male factor. These studies address several aspects: endocrine changes, abnormal sperm, transmission of hereditary diseases, or the need for testosterone replacement therapy in older age. Regarding the histological changes of aging testes, they have conducted several studies for decades, but have left some issues unclear and have focused on humans and rats. Alongside this, in recent years, several authors have proposed that the mechanism involved in decrease of spermatogenesis with age, is related to changes in the balance between proliferation / apoptosis in the seminiferous epithelium. In this thesis we have proposed on the one hand, a complete histological analysis of qualitative and quantitative changes that occur with age in the seminiferous epithelium and tubular wall of the Syrian hamster. Furthermore, we have also investigated whether this proliferation in the epithelium altered is linked with the histological changes observed in it. To do this, we use male hamsters 6, 12 and 24 months old. The testes were fixed and processed for conventional light microscopy, immunohistochemistry and transmission electron microscopy. A semiquantitative histological study of sections of seminiferous tubules for determining the degree of gradual deterioration with age, in its epithelium was performed with age. Immunoreactivity to laminin and fibronectin in the tubular wall and proliferating cells was identified. Morphometrically a series of testicular parameters and the total volume of fibronectin and laminin immunoreactivity was calculated. The total proliferation index as the number of proliferating cells by histologic type of tubular section was calculated. Finally, with ultrathin sections, an ultrarestructural description of seminiferous epithelium, and thickness determination of tubular wall was performed. In aged animals a significant increase in the tubular sections with various disorders especially hypospermatogenesis of seminiferous epithelium and arrest of spermatogenesis was observed. The testicular volume, diameter and volume as epithelial volume of seminiferous tubule suffered a significant decrease with age, on the contrary an increased tubule length was observed. An increase in fibronectin and laminin volume in old animals and less proliferation rate in these animals was determined. The number of proliferating cells per histologic section of seminiferous type tubule, progressively and significantly decrease in altered sections respect to normal sections. No significant differences respect to age between histological types sections of seminiferous tubule was observed. Ultrastructurally important cytological changes were observed in the Sertoli cell and the process of spermiogenesis. The wall thickening showed an increase significant in normal, hipospermatogenic and spermatogenesis arrest tubules. The number of proliferating cells per histologic section of seminiferous type tubule, progressively and significantly decrease in altered sections respect to normal sections. In conclusion, the aging of the seminiferous tubule hamster suppose a loss of spermatogenic activity which is related to changes in the tubule and its wall. These changes do not occur uniformly throughout the tubule and they associated with a decrease in cell proliferation in altered tubular areas. These areas also have Sertoli cells with a modified ultrastructure.

Syllis Savigny in Lamarck, 1818, the type genus of Syllidae, is the largest and most diverse within the family. This genus presents many taxonomic, biogeographic, and phylogenetic problems due mainly to the lack of molecular data and morphological synapomorphies for many of the species, but also due to poor or inadequate descriptions. In order to improve the knowledge of the genus, we have undertaken a morphological revision of Australian species, based on type material and material collected over years by the Australian Museum. This is the third paper dealing with this family. Eighteen species are herein described, of which five are new and twelve are redescriptions of already known species, with four new combinations: Syllis imajimai, n. sp.; S. narranuk, n. sp.; S. noolinga, n. sp.; S. similisunzima, n. sp.; S. thylacine, n. sp.; S. busseltonensis (Hartmann-Schröder, 1982) n. comb.; S. patriciae (Hartmann-Schröder, 1981) n. comb.; S. pharobroomensis (Hartmann-Schröder, 1979) n. comb., S. pharynxcircunfusata (Hartmann-Schröder, 1979) n. comb, S. armillaris (O. F. Müller, 1771); S. augeneri Haswell, 1920; S. benelihauae (Campoy & Alquézar, 1982); S. hyalina Grube, 1863; S. cf. nigrescens Grube, 1878; S. profunda Cognetti, 1955; S. truncata Haswell, 1920; and S. variegata Grube, 1860.

Aim: The latitudinal biodiversity gradient is considered a first-order biogeographical pattern for most taxonomic groups. Latitudinal variation in plant diversity is not always consistent, and this could be related to the particular characteristics of different forest types. In this study, we compare latitudinal changes in floristic diversity (alpha diversity), composition (beta diversity) and dominance across different tropical forest types: floodplain, terra firme and submontane forests. Location: Western Amazonia (Ecuador, Peru and Bolivia). Taxon: Woody plants. Methods: We inventoried 1978 species and 31,203 individuals of vascular plants with a diameter at breast height ≥ 2.5 cm in 118 0.1-ha plots over an 1800 km latitudinal gradient in three different forest types. The relationships between alpha diversity, latitude and forest type were analysed using generalised linear mixed models. Semi-parametric permutational multivariate analysis of variance was used to investigate the effects of latitude and forest type on beta diversity. Dominant species abundances were correlated with non-metric multidimensional scaling ordination axes to reflect their contributions in shaping changes in beta diversity. Results: Alpha diversity increased towards equatorial latitudes in terra firme and submontane forests but remained relatively constant in floodplains. Beta diversity of all forest types changed with latitude, although less clearly in floodplains. Also, in floodplain forests, there were fewer dominant species contributing to beta diversity and more species homogeneous along the gradient. Main Conclusions: Latitudinal diversity patterns are manifested in alpha and beta diversity since latitude summarizes climatic and edaphic changes. However, we found different responses of each forest type. In floodplain forests, inundation regime is a stronger predictor than latitude, limiting floristic diversity and composition. Changes in dominant species abundance over gradients explained species composition, but floodplain forests harboured more homogeneous dominant species than well drained forests. It is key to study environmental trends and habitat characteristics of each forest type to understand their species diversity and dominance patterns

The 10boron neutron-capture therapy (BNCT) is an emerging antitumoral method that shows increasing biomedical interest. BNCT is based on the selective accumulation of the 10boron isotope within the tumor, which is then irradiated with low-energy thermal neutrons, generating nuclear fission that produces 7lithium, 4helium, and γ rays. Simple catechol-borate esters have been rather overlooked as precursors of melanin biosynthesis, and therefore, a proof-of-concept approach for using dopamine-borate (DABO) as a suitable boron-containing candidate for potential BNCT is presented here. DABO can spontaneously oxidize and autopolymerize in vitro, giving a soluble, eumelanin-like brown-black poly-DABO product. Melanotic melanoma cell cultures treated with 1 mM DABO for 24 and 48 h were viable and showed no signs of damage or cell death. The stability and possible trans-esterification of DABO is shortly discussed. Chemical calculations and quantitative structure-activity relationships (QSAR) analysis of DABO and the BNCT agent BPA indicated that they should be cell permeant and accumulate within lysosomes and melanosomes. Molecular modeling allows visualization of both the DABO precursor and the structure of a borate derivative of the proposed catechol-porphycene model for eumelanin, showing interesting features from molecular orbital calculations. The main difference between DABO and other agents, such as BPA, is that it is not a boronic acid nor a boron cluster. This simple catechol-borate ester (protected from oxidation and blackening) could be administrated to living cells and organisms, in which biosynthesis of boron-melanin in melanoma melanocytes can lead to improved BNCT, These authors received no specific funding for this study

The knowledge and control of invasive species are essential in the management of marine ecosystems. The Mediterranean Sea is one of the marine areas with the highest number of invasive species detected, most of them invertebrates. On a smaller scale, colonization by non-native species in coastal lagoons depends directly on their connectivity with the adjacent sea and on the changes in their environmental conditions induced mainly by human interventions. In this study, the cirratulid species Timarete caribous Grube, 1859, native from the Western and Central Atlantic Ocean, was detected for the first time in a Western Mediterranean coastal lagoon, the Mar Menor. It has been found in dense aggregates on both artificial and natural rocky substrates, mainly in the most important communication channel with the Mediterranean Sea. Following the evolution of this NIS will be important for understanding the dynamic and spread of the species and its possible effects on the indigenous biological communities

Mismanaged plastic litter submitted to environmental conditions may breakdown into smaller fragments, eventually reaching nano-scale particles (nanoplastics, NPLs). In this study, pristine beads of four different types of polymers, three oil-based (polypropylene, PP; polystyrene, PS; and low-density polyethylene, LDPE) and one bio-based (polylactic acid, PLA) were mechanically broken down to obtain more environmentally realistic NPLs and its toxicity to two freshwater secondary consumers was assessed. Thus, effects on the cnidarian Hydra viridissima (mortality, morphology, regeneration ability, and feeding behavior) and the fish Danio rerio (mortality, morphological alterations, and swimming behavior) were tested at NPLs concentrations in the 0.001 to 100 mg/L range. Mortality and several morphological alterations were observed on hydras exposed to 10 and 100 mg/L PP and 100 mg/L LDPE, whilst regeneration capacity was overall accelerated. The locomotory activity of D. rerio larvae was affected by NPLs (decreased swimming time, distance or turning frequency) at environmentally realistic concentrations (as low as 0.001 mg/L). Overall, petroleum- and bio-based NPLs elicited pernicious effects on tested model organisms, especially PP, LDPE and PLA. Data allowed the estimation of NPLs effective concentrations and showed that biopolymers may also induce relevant toxic effects, The authors acknowledge the financial support provided by the Spanish Government (Ministerio de Ciencia e Innovación): PID2020-113769RBC21/22, PLEC2021-007693 and the Thematic Network of Micro- and Nanoplastics in the Environment (RED2018-102345-T, EnviroPlaNet Network) and by the Portuguese Government: Project NanoPlanet (2022.02340.PTDC) and financial support to CESAM (UIDP/ 50017/ 2020+UIDB/50017/2020+LA/P/0094/2020), through FCT/MCTES. MTB is the recipient of a FPU (FPU17/01789) pre-doctoral contract by the Spanish Ministerio de Ciencia, Innovación y Universidades

Rice is one of the most important crops in the world and is considered a strategic crop for food security. Furthermore, the excessive use of chemical fertilizers to obtain high yields causes environmental problems. A sustainable alternative includes taking advantage of beneficial bacteria that promote plant growth. Here, we investigate the effect of five bacterial biofertilizers from halophytes on growth, and we investigate photosynthetic efficiency in rice plants grown under saline conditions (0 and 85 mmol L−1 NaCl) and future climate change scenarios, including increased CO2 concentrations and temperature (400/700 ppm and 25/+4 °C, respectively). Biofertilizers 1–4 increased growth by 9–64% in plants grown with and without salt in both CO2- temperature combinations, although there was no significant positive effect on the net photosynthetic rate of rice plants. In general, biofertilizer 1 was the most effective at 400 ppm CO2 and at 700 ppm CO2 +4 °C in the absence of salt. Inocula 1–5 also stimulated plant length at high CO2 levels without salt. Finally, the positive effect of biofertilization was attenuated in the plants grown under the interaction between salt and high CO2. This highlights the significance of studying biofertilization under stress interaction to establish the real potential of biofertilizers in the context of climate change conditions.

Most experimental approaches commonly employed for the characterization and quantitation of EVs are time consuming, require of specialized instrumentation and often are rather inaccurate. To circumvent the caveats imposed by EV small size, we used general and specific membrane markers in bead assisted flow cytometry, to provide a semi-quantitative measure of EV content in a given sample. EVs were isolated from in vitro cultured cells-conditioned medium and biological fluids by size exclusion chromatography and coupled to latex beads to allow their detection by standard flow cytometers. Our analyses demonstrate a linear correlation between EV concentration and Mean Fluorescence Intensity values in samples cleared of protein contaminants. Comparison with one of the most widespread method such as NTA, suggests a similar linear range and reliable accuracy to detect saturation. However, although detection of the different biomarkers is feasible when tested on ultracentrifugation-enriched samples, protein contamination impairs quantitation of this type of samples by bead-based flow cytometry. Thus, we provide evidence that bead-assisted flow cytometry method is an accurate and reliable method for the semiquantitative bulk analysis of EVs, which could be easily implemented in most laboratories, This work was supported by grants BFU2014-55478-R from Ministerio de Economía y Competitividad, and from Fundación Ramón Areces to MY-M; grants PI13/00050 from the “Fondo de Investigación Sanitaria” (FISISCIII), 2014SGR804 from Secretaria d’Universitats i Recerca del Departament d’Economia i Coneixement de la“Generalitat de Catalunya” and REDinREN (16/0009 Feder Funds) to FEB; BFU2014-54181-P from Ministerio de Economía y Competitividad to JLC, and by SAF2016-77096-R from Ministerio de Economía y Competitividad to CC. H.S. was supported by a FPI-UAM and GEIVEX Mobility fellowship; AGV is sponsored by a Grant (482/U/2014) from Fundació La Marató TV3; FEB is sponsored by the “Researchers Stabilization Program” from the Spanish “Sistema Nacional de Salud” (SNS-ISCIII) and “Direcció d’Estratègia i Coordinació”, Catalan Health Dept. (CES07/015) and MY-M by Ramón y Cajal contract RYC-2012-11025

Glycerol fermentation for 1,3-propanediol (1,3-PDO) production was studied in an anaerobic up-flow reactor with biomass attached to silicone support. A mixed microbial culture was activated to perform the biofilm formation and attachment to a silicone hose prior the reactor operation. The reactor was operated over 362 days divided into two phases (P). In P1 and P2, the reactor was continuously fed with pure and crude glycerol, respectively. The operation consisted of increasing the glycerol loading rates (gly-LR). The achieved highest 1,3-PDO average yields were 0.43 mol mol-gly−1 and 0.62 mol mol-gly−1 when applied gly-LR was 18 and 46 g L−1 d−1 in P1 and P2, respectively. A maximum 1,3-PDO productivity of 14.7 g L−1 d−1 was obtained in P2. The higher yields of 1,3-PDO when the feed changed from pure to crude glycerol indicated a change in the microbial community. These results show that the studied system can be very promising and cost-effective for converting crude glycerol into value-added products on large scale even at high loads, This study was funded by Spanish Ministry of Science and Innovation (Grant no: PID2019-104812GBI00X), UAM-Santander Inter-university Cooperation Projects with Latin America (CEAL-AL/2017-14), and CAPES-PrInt (fnance code 001, Process no 88887.467533/2019-00)

Accurate assessment of predation risk is critical for prey survival during predator–prey interactions. Prey can assess predation risk by the presence of cues dropped by predators themselves, but they can also gather information about risk level through cues released by other prey, avoiding the hazard of being in close proximity to predators. In this study, we examine the ability of anuran larvae (Pelobates cultripes) to detect predation risk indirectly when they are in contact with conspecifics that have been recently exposed to chemical stimuli from natural predators (larvae of aquatic beetles). In a first experiment, we confirmed that larvae exposed to predator cues exhibited innate defensive behavior, indicating that they perceived the risk of predation and, thus, could potentially act as risk indicators for naïve conspecifics. In a second experiment, we observed that unexposed larvae paired with a startled conspecific adjusted their antipredator behavior, presumably by mirroring conspecifics’ behavior and/or using chemical cues from their partners as a risk information source. This cognitive ability of tadpoles to assess predation risk through conspecific cues might play an important role in their interaction with predators, facilitating the early detection of potential threats to elicit appropriate antipredator responses and increase the chances of survival, Financial support was provided by the Spanish Ministry of Science and Innovation (MICINN), under Grant CGL2015-68670-R

Obtaining cell concentration measurements from a culture assay by using bioimpedance is a very useful method that can be used to translate impedances to cell concentration values. The purpose of this study was to find a method to obtain the cell concentration values of a given cell culture assay in real time by using an oscillator as the measurement circuit. From a basic cell–electrode model, enhanced models of a cell culture immersed in a saline solution (culture medium) were derived. These models were used as part of a fitting routine to estimate the cell concentration in a cell culture in real time by using the oscillation frequency and amplitude delivered by the measurement circuits proposed by previous authors. Using real experimental data (the frequency and amplitude of oscillations) that were obtained by connecting the cell culture to an oscillator as the load, the fitting routine was simulated, and real-time data of the cell concentration were obtained. These results were compared to concentration data that were obtained by using traditional optical methods for counting. In addition, the error that we obtained was divided and analyzed in two parts: the first part of the experiment (when the few cells were adapting to the culture medium) and the second part of the experiment (when the cells exponentially grew until they completely covered the well). Low error values were obtained during the growth phase of the cell culture (the relevant phase); therefore, the results obtained were considered promising and show that the fitting routine is valid and that the cell concentration can be measured in real time by using an oscillator.

Tau is a well-known microtubule-associated protein related to its cytoplasmic localization in a neuronal cell. However, tau has been located at the cell nucleus where it could be a nucleic acid-associated protein by its preferential binding to DNA sequences present in the nucleolus and pericentromeric heterochromatin. This less well-known localization of tau could not be trivial, since during aging, an increase in the amount of nuclear tau takes place and it may be related to the described role of tau in the activation of transposons and further aging acceleration, Work in the laboratory of JA is funded by grants from the Spanish Ministry of Economy and Competitiveness (PGC-2018-09177-B-100). Work in the laboratory of FH is funded by grants from the Spanish Ministry of Economy and Competitiveness (Ministerio de Economía, Industria y Competitividad, Gobierno de España, PID2020-113204GB-I00) and was co-financed from the Comunidad de Madrid through Structural Funds of the European Union [S2017/BMD-3700 (NEUROMETAB-CM)]

The emergent human coronavirus SARS-CoV-2 and its resistance to current drugs makes the need for new potent treatments for COVID-19 patients strongly necessary. Dextran sulfate (DS) polysaccharides have long demonstrated antiviral activity against different enveloped viruses in vitro. However, their poor bioavailability has led to their abandonment as antiviral candidates. Here, we report for the first time the broad-spectrum antiviral activity of a DS-based extrapolymeric substance produced by the lactic acid bacterium Leuconostoc mesenteroides B512F. Time of addition assays with SARS-CoV-2 pseudoviruses in in vitro models confirm the inhibitory activity of DSs in the early stages of viral infection (viral entry). In addition, this exopolysaccharide substance also reports broad-spectrum antiviral activity against several enveloped viruses such as SARS-CoV-2, HCoV229E, HSV-1, in in vitro models and in human lung tissue. The toxicity and antiviral capacity of DS from L. mesenteroides was tested in vivo in mouse models which are susceptible to SARS-CoV-2 infection. The described DS, administered by inhalation, a new route of administration for these types of polymers, shows strong inhibition of SARS-CoV-2 infection in vivo, significantly reducing animal mortality and morbidity at non-toxic doses. Therefore, we suggest that it may be considered as a potential candidate for antiviral therapy against SARS-CoV-2, Financial support for the study was provided by the REACT-EU 2021 grant from Comunidad de Madrid to the Project COVTRAVI19-CM, Plataformas y modelos preclínicos para el abordaje multidisciplinar en COVID-19 y en respuesta a futuras pandemias

The hollow protein capsids from a number of different viruses are being considered for multiple biomedical or nanotechnological applications. In order to improve the applied potential of a given viral capsid as a nanocarrier or nanocontainer, specific conditions must be found for achieving its faithful and efficient assembly in vitro. The small size, adequate physical properties and specialized biological functions of the capsids of parvoviruses such as the minute virus of mice (MVM) make them excellent choices as nanocarriers and nanocontainers. In this study we analyzed the effects of protein concentration, macromolecular crowding, temperature, pH, ionic strength, or a combination of some of those variables on the fidelity and efficiency of self-assembly of the MVM capsid in vitro. The results revealed that the in vitro reassembly of the MVM capsid is an efficient and faithful process. Under some conditions, up to ~40% of the starting virus capsids were reassembled in vitro as free, non aggregated, correctly assembled particles. These results open up the possibility of encapsidating different compounds in VP2-only capsids of MVM during its reassembly in vitro, and encourage the use of virus-like particles of MVM as nanocontainers, This research was funded by grants from MICINN/FEDER EU (Spain, RTI2018-096635-B-100 and PID2021-126973OB-I00) to M.G.M. and by an institutional grant from Fundación Ramón Areces

In eutherian mammals, hundreds of programmed DNA double-strand breaks (DSBs) are generated at the onset of meiosis. The DNA damage response is then triggered. Although the dynamics of this response is well studied in eutherian mammals, recent findings have revealed different patterns of DNA damage signaling and repair in marsupial mammals. To better characterize these differences, here we analyzed synapsis and the chromosomal distribution of meiotic DSBs markers in three different marsupial species (Thylamys elegans, Dromiciops gliorides, and Macropus eugenii) that represent South American and Australian Orders. Our results revealed inter-specific differences in the chromosomal distribution of DNA damage and repair proteins, which were associated with differing synapsis patterns. In the American species T. elegans and D. gliroides, chromosomal ends were conspicuously polarized in a bouquet configuration and synapsis progressed exclusively from the telomeres towards interstitial regions. This was accompanied by sparse H2AX phosphorylation, mainly accumulating at chromosomal ends. Accordingly, RAD51 and RPA were mainly localized at chromosomal ends throughout prophase I in both American marsupials, likely resulting in reduced recombination rates at interstitial positions. In sharp contrast, synapsis initiated at both interstitial and distal chromosomal regions in the Australian representative M. eugenii, the bouquet polarization was incomplete and ephemeral, γH2AX had a broad nuclear distribution, and RAD51 and RPA foci displayed an even chromosomal distribution. Given the basal evolutionary position of T. elegans, it is likely that the meiotic features reported in this species represent an ancestral pattern in marsupials and that a shift in the meiotic program occurred after the split of D. gliroides and the Australian marsupial clade. Our results open intriguing questions about the regulation and homeostasis of meiotic DSBs in marsupials. The low recombination rates observed at the interstitial chromosomal regions in American marsupials can result in the formation of large linkage groups, thus having an impact in the evolution of their genomes, This work was supported by grants CGL 2014-53106-P to JP (Ministerio de Ecomonía y Competitividad, Spain), BIOUAM02- 2020 to JP and RG (Departamento de Biología, Universidad Autónoma de Madrid), PID 2020-112557 GB-I00 to AR-H. (Ministerio de Ciencia e Innovación, Spain) and from the Australian Research Council to MBR, GS, and PDW. (DP21103512 and DP220101429). PW is also supported by the NHMRC (APP1182667 and APP2021172). LM-G was supported by a FPU predoctoral fellowship from the Ministry of Science, Innovation and University (FPU18/03867)

Background: Alzheimer’s disease (AD) and type 2 diabetes mellitus (T2DM) share metabolic alterations such as abnormal insulin and lipid metabolism and have some common genetic factors such as APOE genotype. Taking this into account, we hypothesized that we could identify common genetic factors involved in the development of diabetes and cardiovascular diseases. Methodology: We first genotyped 48 single nucleotide polymorphisms (SNPs) previously associated with AD in a cohort composed of 330 patients with cognitive impairment (CI) to assess their association with plasma lipids. Second, we conducted pleiotropy-informed conjunctional false discovery rate (FDR) analysis designed to identify shared variants between AD and plasma lipid levels. Finally, we used the SNPs to be found associated with lipid parameters and AD to search for associations with lipoprotein parameters in 281 patients with cardiometabolic risk. Results: Five SNPs were significantly associated with lower levels of cholesterol transported in remnant lipoprotein particles (RLPc) in subjects with CI; among these SNPs was the rs73572039 variant in PVRL2. Stratified QQ-plots were conducted on GWAS designed for AD and triglycerides (TG). The cross-trait analysis resulted in a total of 22 independent genomic loci associated with both AD and TG levels with a conjFDR < 0.05. Among these loci, two pleiotropic variants were located in PVRL2 (rs12978931 and rs11667640). The three SNPs in PVRL2 were significantly associated with RLPc, TG, and number of circulating VLDL and HDL particles in subjects with cardiometabolic risk. Conclusions: We have identified three variants in PVRL2 that predispose individuals to AD that also influence the lipid profile that confers cardiovascular risk in T2DM subjects. PVRL2 is a potential new modulating factor of atherogenic dyslipidemia, This work was supported by a research grant from the Spanish Ministerio de Ciencia, Innovación y Universidades (PI19/00832), Fondo Europeo de Desarrollo Regional (FEDER), and CIBERDEM (CIBER de Diabetes y Enfermedades Metabólicas Asociadas), which are initiatives of Instituto de Salud Carlos III (ISCIII); by Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR) (2017SGR0444); and by the Cerca Programme, Generalitat de Catalunya

A limited number of effective therapies are currently available to treat human coronavirus SARS-CoV-2 and other human coronaviruses, which are responsible for nearly a third of global cases of the common cold. The possibility of new emerging coronaviruses demands powerful new antiviral strategies. Lactoferrin is a well-known protein that possesses anti-inflammatory and immunomodulatory activities, and it has previously shown antiviral activity against several viruses, including SARS-CoV-2. To increase this antiviral activity, here we present bovine liposomal lactoferrin. Liposomal encapsulation of the compound was proven to increase permeability, bioavailability, and time release. In the present work, we compare the antiviral activity of free and liposomal bovine lactoferrin against HCoV229E and SARS-CoV-2 in vitro and in human primary bronchial epithelial cells, and we demonstrated that the liposomal form exerts a more potent antiviral activity than its free form at non-cytotoxic doses.

The species Coniopteris laciniata and Sphenopteris wonnacottii are heterophyllous ferns recovered from two Spanish Barremian localities, Las Hoyas and El Montsec respectively. The similarities between these species, observed in a study of a total of 66 hand specimens from both localities, indicate that they are conspecific. Following the rule of priority in botanical nomenclature, and because there are doubts about the assignment of this species to a higher taxonomic rank, the name Coniopteris is maintained. Characters of this fern and additional characters of other species of Coniopteris would support a diversification of polypod ferns during the Early Cretaceous. Metric architecture analyses suggest that the heterophylly of the plant was probably caused by the submersion of the apical part of fronds in water during their development, which would be consistent with the taphonomy of the remains in both localities, We would like to thank El Montsec housed at the Muséum national d’Histoire naturelle (Paris, France), which was visited with support of a grant from the SYNTHESYS Project (http://www.synthesys.info/), financed by the European Community Research Infrastructure Action under the FP7 Integrating Activities Program. Candela Blanco-Moreno is supported by a Margarita Salas Postdoc CA1/RSUE/2021-00703 scholarship funded by the Spanish Ministry of Universities and Universidad Autónoma de Madrid. This work is part of the project PID2019-105546GB-I00 of the Spanish Ministry of Science, Innovation and Universities

Leishmania infantum is one of the causative agents of visceral leishmaniases, the most severe form of leishmaniasis. An improved assembly for the L. infantum genome was published five years ago, yet delineation of its transcriptome remained to be accomplished. In this work, the transcriptome annotation was attained by a combination of both short and long RNA-seq reads. The good agreement between the results derived from both methodologies confirmed that transcript assembly based on Illumina RNA-seq and further delimitation according to the positions of spliced leader (SAS) and poly-A (PAS) addition sites is an adequate strategy to annotate the transcriptomes of Leishmania, a procedure previously used for transcriptome annotation in other Leishmania species and related trypanosomatids. These analyses also confirmed that the Leishmania transcripts boundaries are relatively slippery, showing extensive heterogeneity at the 5′- and 3′-ends. However, the use of RNA-seq reads derived from the PacBio technology (referred to as Iso-Seq) allowed the authors to uncover some complex transcription patterns occurring at particular loci that would be unnoticed by the use of short RNA-seq reads alone. Thus, Iso-Seq analysis provided evidence that transcript processing at particular loci would be more dynamic than expected. Another noticeable finding was the observation of a case of allelic heterozygosity based on the existence of chimeric Iso-Seq reads that might be generated by an event of intrachromosomal recombination. In addition, we are providing the L. infantum gene models, including both UTRs and CDS regions, that would be helpful for undertaking whole-genome expression studies. Moreover, we have built the foundations of a communal database for the active curation of both gene/transcript models and functional annotations for genes and proteins, This research was supported by the Spanish Ministerio de Ciencia, Innovación (MICINN), Agencia Estatal deInvestigación(AEI), grant number PID2020-117916RB-I00, and Instituto de Salud Carlos III, grant CB21/13/00018 (CIBERINFEC). An institutional grant from Fundacion RamonAreces is also acknowledged

11 Páginas.-- 1 Figura.-- 5 Tablas.-- Material suplementario, Hemp is the term commonly used to refer to the variety of Cannabis sativa L. cultivated for industrial purposes. The seeds have gained interest in recent years as functional foods due to their nutritional composition and high content of protein and bioactive compounds. In this study, ten hemp protein hydrolysates (HPHs) were obtained by enzymatic hydrolysis with Alcalase and Flavourzyme from hemp protein isolate (HPI) and their antioxidant properties (DPPH radical scavenging activity, beta-carotene activity and ferric ion reducing antioxidant power (FRAP)) were evaluated. Shorter peptide sequences, mainly obtained with Flavourzyme, were found to react with free radicals more easily. The peptidome of all the hydrolysates was characterized, identifying, and quantifying the peptides. Furthermore, 19 unique peptides were assessed by in silico tools to hypothesize those that could be responsible of the bioactivity reported for the hydrolysates. From the identified peptides, based on the molecular features and the predictions, the peptides KNAIYTPH, EERPGHF, and KNGMMAPH, among others, are proposed to be highly contributing to the antioxidant activity of the hydrolysates., This research was funded by grant P20_00661 from the Andalusian Plan for Research, Development and Innovation (PAIDI) 2020 and by grant US-1381492 from the European Regional Development Fund (ERDF).

The hippocampus hosts the continuous addition of new neurons throughout life—a phenomenon named adult hippocampal neurogenesis (AHN). Here we revisit the occurrence of AHN in more than 110 mammalian species, including humans, and discuss the further validation of these data by single-cell RNAseq and other alternative techniques. In this regard, our recent studies have addressed the long-standing controversy in the field, namely whether cells positive for AHN markers are present in the adult human dentate gyrus (DG). Here we review how we developed a tightly controlled methodology, based on the use of high-quality brain samples (characterized by short postmortem delays and ≤24 h of fixation in freshly prepared 4% paraformaldehyde), to address human AHN. We review that the detection of AHN markers in samples fixed for 24 h required mild antigen retrieval and chemical elimination of autofluorescence. However, these steps were not necessary for samples subjected to shorter fixation periods. Moreover, the detection of labile epitopes (such as Nestin) in the human hippocampus required the use of mild detergents. The application of this strictly controlled methodology allowed reconstruction of the entire AHN process, thus revealing the presence of neural stem cells, proliferative progenitors, neuroblasts, and immature neurons at distinct stages of differentiation in the human DG. The data reviewed here demonstrate that methodology is of utmost importance when studying AHN by means of distinct techniques across the phylogenetic scale. In this regard, we summarize the major findings made by our group that emphasize that overlooking fundamental technical principles might have consequences for any given research field, Association for Frontotemporal Degeneration; Banco de Santander; Center for Networked Biomedical Research on Neurodegenerative Diseases; Consejo Nacional de Ciencia y Tecnología (CONACYT), Grant/Award Number: 385084; European Research Council, Grant/Award Number: ERC-CoG2020-101001916; Fundacion Ram on Areces; Secretaria de Educacion, Ciencia Tecnología e Innovacion (SECTEI) of the Regional Government of Ciudad de México (CDMX), Grant/Award Number: SECTEI/159/2021; Spanish Ministry of Economy and Competitiveness, Grant/Award Numbers: PID2020-113007RB-I00, RYC-2015-171899, SAF-2017-82185-R; The Alzheimer's Association, Grant/Award Numbers: 2015-NIRG-340709, AARG-17-528125, AARG-17-528125-RAPID

Among the extracellular vesicles, apoptotic bodies (ABs) are only formed during the apoptosis and perform a relevant role in the pathogenesis of different diseases. Recently, it has been demonstrated that ABs from human renal proximal tubular HK-2 cells, either induced by cisplatin or by UV light, can lead to further apoptotic death in naïve HK-2 cells. Thus, the aim of this work was to carry out a non-targeted metabolomic approach to study if the apoptotic stimulus (cisplatin or UV light) affects in a different way the metabolites involved in the propagation of apoptosis. Both ABs and their extracellular fluid were analyzed using a reverse-phase liquid chromatography-mass spectrometry setup. Principal components analysis showed a tight clustering of each experimental group and partial least square discriminant analysis was used to assess the metabolic differences existing between these groups. Considering the variable importance in the projection values, molecular features were selected and some of them could be identified either unequivocally or tentatively. The resulting pathways indicated that there are significant, stimulus-specific differences in metabolites abundancies that may propagate apoptosis to healthy proximal tubular cells; thus, we hypothesize that the share in apoptosis of these metabolites might vary depending on the apoptotic stimulus, This research was funded by the Spanish Ministry of Science and Innovation (project PID2019-104913GB-I00, Agencia Estatal de Investigación, Referencia del Proyecto/AEI/10.13039/ 501100011033) and the Community of Madrid, I+D Biomedicine Activities Program 2022 (General Direction of Research and Technological Innovation (project P2022/BMD7223 CIFRA_CORCM)

Zinnia elegans constituye uno de los sistemas modelos más útiles para estudiar la diferenciación en el xilema que incluye la síntesis de la pared celular secundaria, la lignificación de la pared celular, y la muerte celular programada. Además, el cultivo de células del mesófilo de Z. elegans in vitro también se ha utilizado como sistema modelo ya que la diferenciación de las células del mesófilo en traqueidas permite estudiar de manera sencilla la bioquímica y la fisiología de la xilogénesis alejados de la complejidad que imponen los tejidos vegetales. Por otra parte, Z. elegans ha resultado ser una excelente planta modelo para estudiar la implicación de las peroxidasas en la lignificación de la pared celular. Esto es debido a la simplicidad y dualidad del patrón de lignificación mostrado en los tallos e hipocotilos de esta planta y a la naturaleza básica de esta isoenzima de peroxidasa. Sin embargo, esta proteína no solo se expresa en hipocotilos y en tallos sino también en células que se encuentran en diferenciación en traqueidas. Por lo tanto, esta peroxidasa no solo cumple todos los requerimientos catalíticos impuestos para ser la candidata idónea implicada en la lignificación cumpliendo con todas las restricciones impuestas por la etapa de acoplamiento oxidativo de los alcoholes hidroxicinamilicos y polimerización, sino también debido a que su expresión es inherente en la lignificación. De hecho, su naturaleza básica no es excepcional ya que las peroxidasas básicas se expresan diferencialmente durante la lignificación en otros sistemas modelo, mostrando propiedades bioquímicas inusuales y exclusivas tales como la oxidación de las unidades siringilo. Esta Tesis se ha centrado en la realización de las actividades que conducen a un mejor entendimiento de la lignificación en Zinnia, comenzando con el estudio del patrón de lignificación, cómo se sucede este proceso, y la implicación de la peroxidasa, demostrando sus propiedades catalíticas inusuales y cómo esta enzima es regulada por el óxido nítrico (NO) y el H2O2. En relación a la producción de NO y H2O2 en los haces vasculares, utilizando la microscopía confocal láser de barrido, se observó un gradiente espacial de producción de NO inversamente relacionado con el grado de diferenciación del xilema ya que las células del xilema jóvenes, en diferenciación manifestaron una mayor capacidad para producir NO que las células del xilema completamente maduras y diferenciadas. Estos resultados demuestran la fuerte relación entre la producción de NO y la muerte celular programada que se produce durante la diferenciación del xilema y que precede a la lignificación de la pared celular. Además, la producción de NO en los cultivos celulares de mesófilo de Z. elegans que se diferencian a traqueidas, las células del mesófilo mantienen la misma propiedad mostrada por las células del xilema en diferenciación ya que la señal fluorescente se detectó, principalmente, en las células predeterminadas y en las que se encontraban en diferenciación. Por el contrario, las células aisladas del mesófilo que no habían adquirido la capacidad para diferenciarse a traqueidas, manifestaban unos niveles muy débiles de la señal fluorescente que contrastaba con la producción elevada mostrada por las células predeterminadas para diferenciarse a traqueidas. Por otra parte, mediante la utilización conjunta de técnicas de microscopía óptica y electrónica, se realizó un estudio detallado de los sitios de producción de H2O2 en el xilema en lignificación, comprobándose que en las células del xilema en diferenciación y en las células del parénquima adyacentes se localizaba la producción de H2O2 sugiriendo que estas células podrían ser las responsables de la producción de H2O2 que, posteriormente, difunde de célula a célula, a través del espacio intercelular hacia las células del xilema en diferenciación para que tenga lugar la biosíntesis de ligninas en la pared celular secundaria. Una evidencia adicional que apoyó la hipótesis planteada se obtuvo mediante la localización del H2O2 en los cultivos celulares de mesófilo de Z. elegans en diferenciación a elementos traqueales. Utilizando este sistema modelo junto con la microscopía confocal láser y el diacetato de diclorofluoresceína como sonda fluorescente, la producción de H2O2 se localizó mayoritariamente en las células del mesófilo predeterminadas para diferenciarse, detectándose un bajo nivel de H2O2 en las traqueidas completamente diferenciadas., Zinnia elegans constitutes one of the most useful model systems for studying xylem differentiation, which simultaneously involves secondary cell wall synthesis, cell wall lignification, and programmed cell death. Likewise, the in vitro culture system of Z. elegans has been the best characterized since the differentiation of mesophyll cells into tracheary elements allows study the biochemistry and physiology of xylogenesis free from the complexity that heterogeneous plant tissues impose. Moreover, Z. elegans has resulted in an excellent plant model to study the involvement of peroxidases in cell wall lignification. This has been due to the simplicity and duality of the lignification pattern shown by the stems and hypocotyls, and to the basic nature of the peroxidase isoenzyme. This protein is expressed not only in hypocotyls and stems but also in mesophyll cells transdifferentiating into tracheary elements. Therefore, not only does this peroxidase fulfil all the catalytic requirements to be involved in lignification overcoming all restrictions imposed by the polymerization step, but also its expression is inherent in lignification. In fact, its basic nature is not exceptional since basic peroxidases are differentially expressed during lignification in other model systems, showing unusual and unique biochemical properties such as oxidation of syringyl moieties. This Thesis has focused on the experiments which led to a better understanding of the lignification process in Zinnia, starting with the basic knowledge about its lignin pattern, how lignification takes place, and the involvement of the basic peroxidase in the lignification process, demonstrating its unusual catalytic properties, and how this enzyme is regulated by nitric oxide and H2O2. In relation to the production of NO in the vascular bundles using confocal laser scanning microscopy, a spatial NO gradient inversely related to the degree of xylem differentiation was observed since differentiating xylem cells clearly manifest a higher capacity for NO production than differentiated xylem cells. In addition, the NO production in cultured mesophyll cells which are trans-differentiating in tracheary elements maintain the same property shown by differentiating xylem from the Z. elegans vascular bundles since the fluorescent probe was mainly observed in pro-differentiating thin-walled and differentiating (secondary cell wall forming) tracheary elements. By the contrary, mesophyll cells which have not acquired competence for trans-differentiation showed only background levels of triazolofluorescein green fluorescence. These results confirm that NO formation by tracheary elements is directly related to the early processes of xylem differentiation, which takes place immediately before the late processes of secondary cell wall formation and cell autolysis. On the other hand, a study of both localization and production of H2O2 in the lignifying xylem by the joint use of optical and electronic microscopy and 2,7-dichlorofluorescein diacetate and CeCl3 assays was carried out. Thus, H2O2 production was observed in thin-walled cells surround thick-walled xylem vessels and in the xylem parenchyma cells intercalated between xylem vessels suggesting that these cells could be responsible of H2O2 production and a degree of cell to cell cooperation is produced during lignin biosynthesis. Further evidence that supports this hypothesis was obtained by studying the production and localization of H2O2 during the differentiation of Z. elegans mesophyll cells into tracheary elements. Using this model system and confocal laser scanning microscopy and 2,7-dichlorofluorescein diacetate as fluorescent probe, H2O2 was mainly produced by both differentiating tracheary and parenchyma-like elements being the non-lignifying parenchyma-like cells the main H2O2 source and detecting a low level of H2O2 in the totally differentiated tracheary elements.