Caracterización molecular del metabolismo del alantoato en judía (Phaseolus Vulgaris)

En plantas, la degradación del alantoato es un paso indispensable para el reciclaje de las bases nitrogenadas. Además, en las leguminosas tropicales, como judía y soja, los ureidos alantoína y alantoato son las formas mayoritarias de transporte del nitrógeno fijado y su acumulación se ha relacionado con la inhibición de la fijación de nitrógeno en condiciones de sequía. En este trabajo se ha identificado y caracterizado una alantoato amidohidrolasa (PvAAH) en judía. Los resultados obtenidos tras bloquear mediante RNAi la expresión de esta proteína sugieren que la AAH es la enzima responsable de la degradación del alantoato en judía. El análisis del metabolismo del alantoato en plantas de judía mostró un incremento en el contenido de ureidos en las partes aéreas de la planta durante la etapa reproductiva del desarrollo, tanto en plantas noduladas como en plantas sin nódulos, así como en plantas sometidas a oscuridad. Este incremento de ureidos se correlacionó con el aumento de la actividad alantoinasa en los tallos, regulada a nivel transcripcional, y con el descenso de la actividad PvAAH en las hojas. Además, en ambas situaciones, el aumento en los niveles de ureidos se acompañó con un incremento en los niveles de glucosa y almidón en las hojas. Asimismo, se observó un descenso en la actividad PvAAH en hojas de plantas sometidas a déficit hídrico y un aumento en la expresión del gen PvAAH en respuesta a diferentes hormonas relacionadas con la respuesta a estrés.Allantoate degradation is an essential step for recycling purine-ring nitrogen in all plants. Furthermore, in tropical legumes, as common bean and soybean, the ureides allantoin and allantoate are the mayor transport form of fixed nitrogen and ureide accumulation has been related with the inhibition of nitrogen fixation under drought conditions. In this work, an allantoate amidohydrolase (PvAAH) enzyme from common bean has been identified and characterized. The results obtained after blocking PvAAH expression using RNAi suggest that the PvAAH is the enzyme responsible for degrading allantoate in common bean. The analysis of the allantoate metabolism in common bean plants showed an increase in the ureide content in the aerial tissues during the reproductive developmental stage both, in nodulated and nonoduladed plants, as well as in plants subjected to darkness. This increase of ureides was correlated with an increase of the allantoinase activity in the stems, regulated at transcriptional level, and with a decrease of the PvAAH activity in the leaves. In addition, in both situations, ureide accumulation was accompanied by an increase in glucose and starch in leaves. Likewise, a decrease in the PvAAH activity was observed in leaves of plants submitted to water deficit, and the expression of the PvAAH gene was induced in plants treated with several stress-related phytohormones

Local inhibition of nitrogen fixation and nodule metabolism in drought-stressed soybean

Drought stress is a major factor limiting symbiotic nitrogen fixation (NF) in soybean crop production. However, the regulatory mechanisms involved in this inhibition are still controversial. Soybean plants were symbiotically grown in a split-root system (SRS), which allowed for half of the root system to be irrigated at field capacity while the other half remained water deprived. NF declined in the water-deprived root system while nitrogenase activity was maintained at control values in the well-watered half. Concomitantly, amino acids and ureides accumulated in the water-deprived belowground organs regardless of transpiration rates. Ureide accumulation was found to be related to the decline in their degradation activities rather than increased biosynthesis. Finally, proteomic analysis suggests that plant carbon metabolism, protein synthesis, amino acid metabolism, and cell growth are among the processes most altered in soybean nodules under drought stress. Results presented here support the hypothesis of a local regulation of NF taking place in soybean and downplay the role of ureides in the inhibition of NF

Modelo Prolab: Iron Can, fabricación y comercialización de galletas para perros en base a sangre de ganado vacuno en la región Cajamarca

De acuerdo con la OPS, 2020: “La persistencia y reemergencia de algunas enfermedades infecciosas asociadas a los cambios socioeconómicos y ambientales, ponen de relieve la necesidad de reforzar actividades de la salud pública”, por lo cual la salud pública y el cuidado del medio ambiente es un tema que ha tomado relevancia en las dos últimas décadas, tal es así que actualmente no solo se busca cuidarlo, si no también mejorarlo de manera sostenible, es por ello que en el desarrollo del presente trabajo de tesis se identificó, que dentro de la Ciudad de Cajamarca existe un problema social relevante, el cual es la inadecuada disposición de los residuos generados por el Camal Municipal, causados por el proceso de faenamiento de los bovinos, ya que la sangre es vertida sin tratamiento alguno en los sistemas de drenaje del camal e incluso en algunas ocasiones generan su colapso, ocasionando que las calles aledañas sean cubiertas de sangre lo cual genera un impacto social y ambiental negativo. Para poder obtener una solución sostenible en el tiempo es necesito formar alianzas estratégicas entre la municipalidad, la empresa y la sociedad de manera que permitan mejorar la salud publica mediante la gestión eficiente de los desechos municipales de los camales, dando un valor agregado a más de 70 toneladas anuales de sangre de ganado vacuno que actualmente son vertidas en el drenaje. Es así, que se plantea como solución viable el empleo diario de 250 litros de sangre para la fabricación de galletas para perros. El mercado actual muestra una creciente demanda por la compra de alimentos saludables para mascotas por lo cual se empleó el modelo canvas para estudiar el mercado y medir la rentabilidad del negocio propuesto el cual consiste en utilizar la sangre de ganado vacuno de los camales, la cual es rica en hierro, para la producción y comercialización de galletas para perros. Finalmente, de acuerdo con esto y la simulación de las proyecciones financieras se establece que es un negocio rentable y sostenible, ya que se obtiene un VAN económico de S/. 2,185,509.12 con un TIR de 55% y un VAN financiero de S/. 1,537,983.68 con un TIR de 58%, lo cual permite la rentabilidad del negocio, así como también la liquidez que va a permitir gestionar de manera eficiente desechos contaminantes para la comercialización de las galletas para perro, de igual manera se obtuvo un VAN social de S/ 4,067,009.64 lo que permite establecer un impacto positivo en el medio ambiente, generando un emprendimiento con un objetivo de desarrollo sostenible.According to PAHO, 2020: "The persistence and reemergence of some infectious diseases associated with socioeconomic and environmental changes, highlight the need to strengthen public health activities", for which public health and care for the environment is a topic that has become relevant in the last two decades, so much so that currently it is not only sought to care for it, but also to improve it in a sustainable way, which is why the development of this work of thesis it was identified that within the City of Cajamarca there is a relevant social problem, which is the inadequate disposal of the waste generated by the Municipal Camal, caused by the process of slaughtering the bovines, since at the time of the dumping of The blood in the drainage systems of the slaughterhouse are not treated and on some occasions they even collapse, causing the surrounding streets to be covered in blood, generating a negative social and environmental impact. In order to obtain a sustainable solution over time, it is necessary to form strategic alliances between the municipality, the company and society in order to improve public health through the efficient management of municipal waste from the slaughterhouses, giving added value to more than 70 annual tons of bovine blood that are currently poured into the drain. Thus, the daily use of 250 liters of blood that are produced in the manufacture of dog biscuits is proposed as a viable solution. The current market shows a growing demand for the purchase of healthy pet food, for which the canvas model was used to study the market and measure the profitability of the proposed business, which consists of using the blood of cattle from the slaughterhouses, which It is rich in iron, for the production and sale of biscuits for dogs. Finally, according to this and the simulation of the financial projections, it is established that it is a profitable and sustainable business, since an economic NPV of S/. 2,185,509.12 and an IRR of 55% and a financial NPV of S/. 1,537,983.68 and an IRR of 58%, which allows the profitability of the business, as well as the liquidity that will allow the efficient management of polluting waste for the commercialization of dog biscuits. In the same way, a social NPV of S/ 4,067,009.64 was obtained. which allows establishing the positive impact that it generates in the environment, generating a venture with a sustainable development objective

Multidisciplinary study of mud volcanoes and diapirs and their relationship to seepages and bottom currents in the Gulf of Cádiz continental slope (northeastern sector)

The seabed morphology, type of sediments, and dominant benthic species on eleven mud volcanoes and diapirs located on the northern sector of the Gulf of Cádiz continental slope have been studied. The morphological characteristics were grouped as: (i) fluid-escape-related features, (ii) bottom current features, (iii) mass movement features, (iv) tectonic features and (v) biogenic-related features. The dominant benthic species associated with fluid escape, hard substrates or soft bottoms, have also been mapped. A bottom current velocity analysis allowed, the morphological features to be correlated with the benthic habitats and the different sedimentary and oceanographic characteristics. The major factors controlling these features and the benthic habitats are mud flows and fluid-escape-related processes, as well as the interaction of deep water masses with the seafloor topography. Mud volcano eruptions give rise to mud flows and/or aqueous fluid seepage. These processes sustain chemosynthesis-based communities, closely associated with fluid seepage. Large depressions in the nearby area are influenced by collapse-related phenomena, where active fluid escape and the erosive effect of bottom currents have been identified. When the extrusion activity of the mud volcano is low and the seepage is diffuse, authigenic carbonates form within the edifice sediments. The bottom current sweeps the seafloor from the SE to the NW. When the velocity is moderate, sedimentary contourite processes take place on both sides of the edifices. At high velocities, the authigenic carbonates may be exhumed and colonised by species associated with hard substrates. Small carbonate mounds are found at the summits of some volcanoes and diapirs. Living corals have been found on the tops of the shallowest mud volcanoes, revealing different oceanographic conditions and strong bottom currents that favour the availability of nutrients and organic particles. The edifices affected by very high current velocities are located in the channels where erosive processes dominate.Postprint2,517

PM060184, a new tubulin binding agent with potent antitumor activity including P-glycoprotein over-expressing tumors

PM060184 belongs to a new family of tubulin-binding agents originally isolated from the marine sponge Lithoplocamia lithistoides. This compound is currently produced by total synthesis and is under evaluation in clinical studies in patients with advanced cancer diseases. It was recently published that PM060184 presents the highest known affinities among tubulin-binding agents, and that it targets tubulin dimers at a new binding site. Here, we show that PM060184 has a potent antitumor activity in a panel of different tumor xenograft models. Moreover, PM060184 is able to overcome P-gp mediated resistance in vivo, an effect that could be related to its high binding affinity for tubulin. To gain insight into the mechanism responsible of the observed antitumor activity, we have characterized its molecular and cellular effects. We have observed that PM060184 is an inhibitor of tubulin polymerization that reduces microtubule dynamicity in cells by 59%. Interestingly, PM060184 suppresses microtubule shortening and growing at a similar extent. This action affects cells in interphase and mitosis. In the first case, the compound induces a disorganization and fragmentation of the microtubule network and the inhibition of cell migration. In the second case, it induces the appearance of multipolar mitosis and lagging chromosomes at the metaphase plate. These effects correlate with prometaphase arrest and induction of caspase-dependent apoptosis or appearance of cells in a multinucleated interphase-like state unrelated to classical apoptosis pathways. Taken together, these results indicate that PM060184 represents a new tubulin binding agent with promising potential as an anticancer agent.This work was supported by grants BIO2010-16351 (JFD), CAM S2010/BMD-2457 (JFD), CAM S2010/BMD-2353 (JMA), BFU2011-23416 (JMA) and PharmaMar-CSIC contracts. BP had a contract from Comunidad de Madrid

Generation of organotypic multicellular spheres by magnetic levitation : model for the study of human hematopoietic stem cells microenvironment

Q4Q3Background and Objective: The characteristics of human hematopoietic stem cells are conditioned by the microenvironment of the bone marrow, where they interact with other cell populations, such as mesenchymal stem cells and endothelial cells; however, the study of this microenvironment is complex. The objective of this work was to develop a 3D culture system by magnetic levitation that imitates the microenvironment of human HSC. Methods and Results: Human bone marrow-mesenchymal stem cells, umbilical cord blood-hematopoietic stem cells and a non-tumoral endothelial cell line (CC2811, LonzaⓇ) were used to develop organotypic multicellular spheres by the magnetic levitation method. We obtained viable structures with an average sphericity index greater than 0.6, an average volume of 0.5 mm3 and a percentage of aggregation greater than 70%. Histological studies of the organotypic multicellular spheres used hematoxylin and eosin stains, and an evaluation of vimentin expression by means of immunohistochemistry demonstrated an organized internal structure without picnotic cells and a high expression of vimentin. The functional capacity of human hematopoietic stem cells after organotypic multicellular spheres culture was evaluated by multipotency tests, and it was demonstrated that 3D structures without exogenous Flt3L are autonomous in the maintenance of multipotency of human hematopoietic stem cells. Conclusions: We developed organotypic multicellular spheres from normal human cells that mimic the microenvironment of the human hematopoietic stem cells. These structures are the prototype for the development of complex organoids that allow the further study of the biology of normal human stem cells and their potential in regenerative medicine.https://orcid.org/0000-0002-9152-5552https://orcid.org/0000-0003-3075-9854https://orcid.org/0000-0002-0084-0339https://orcid.org/0000-0003-1881-9367N/

From chemosynthesis-based communities to cold-water corals: Vulnerable deep-sea habitats of the Gulf of Cádiz

The Gulf of Cádiz (GoC) represents an area of ecological importance within the northeastern Atlantic Ocean due to the presence of Mediterranean and Atlantic water masses, a heterogeneous seafloor and a biological confluence. Nevertheless, information on the presence of vulnerable deep-sea habitats is still very scarce and it is of importance for further habitat monitoring within the context of the Habitats and Marine Strategy Framework Directives and for improving conservation and resource extraction management. From 2010 to 2012, fluid migration and emission related edifices (e.g., mud volcanoes, diapirs) from the Spanish continental margin of the GoC have been explored using a remotely operated vehicle (ROV; Liropus 2000) and an underwater camera sled (UCS; APHIA 2012) as well as several devices for collecting sediment and fauna. Different vulnerable deep-sea habitats have been observed, including anoxic bottoms with bacterial mats, sea-pen communities, sponge aggregations, antipatharian and gorgonian communities and also cold-water coral banks. Some of these habitats are included in conservation lists of the habitat directive and in international conventions (OSPAR, RAC/SPA), however some of them are located in areas of the GoC that are exposed to intense trawling. The diversity of habitats detected in the Spanish continental margin of the GoC highlights the importance of seepage related edifices as inducers of seabed and habitat heterogeneity in deep-sea areas.En prens

Geomorphological, oceanographical and benthic characteristics of Gazul mud volcano in the Spanish Middle Slope of the Gulf of Cádiz

The seabed morphology and the sub-bottom characteristics of a sector of the Gulf of Cádiz continental slope have been analyzed from different data sets. The morphological characteristics, the type of sediments and dominant benthic species of Gazul mud volcano and adjacent areas have been studied. The morphological characteristics identified were grouped in fluid escape related features, bottom current features, mass movement features, structural features and biogenic related features. Additionally, dominant benthic species associated to fluid escape and seepage, hard substrates or soft bottoms have also been indicated in these features. A bottom current velocity analysis allowed relating the morphological features with the different sedimentary and oceanographic processes as well as the benthic habitats and that currently occur in the mud volcano and nearby areas. The major factor controlling these features and their associated benthic habitats is the interaction of deep water masses with the seafloor topography. At high velocities, authigenic carbonates can be exhumed and colonized by sessile suspensivorous species associated with hard substrates such as cold-water corals, anthipatarians and large sponges. Mounds and diapiric ridges are found on the depressions and they could be relict structures. Living corals have been found on raised zones, revealing different oceanographic condition and strong enough currents to favour the availability of nutrients and organic particles. Chemosymbiotic organisms are very scarce at the summit and old remains of Lucinoma asapheus indicating past higher seepage activity.Versión del edito

How does neighbourhood socio-economic status affect the interrelationships between functioning dimensions in first episode of psychosis? A network analysis approach

The links between psychosis and socio-economic disadvantage have been widely studied. No previous study has analysed the interrelationships and mutual influences between functioning dimensions in first episode of psychosis (FEP) according to their neighbourhood household income, using a multidimensional and transdiagnostic perspective. 170 patients and 129 controls, participants in an observational study (AGES-CM), comprised the study sample. The WHO Disability Assessment Schedule (WHODAS 2.0) was used to assess functioning, whereas participants' postcodes were used to obtain the average household income for each neighbourhood, collected by the Spanish National Statistics Institute (INE). Network analyses were conducted with the aim of defining the interrelationships between the different dimensions of functioning according to the neighbourhood household income. Our results show that lower neighbourhood socioeconomic level is associated with lower functioning in patients with FEP. Moreover, our findings suggest that “household responsibilities” plays a central role in the disability of patients who live in low-income neighbourhoods, whereas “dealing with strangers” is the most important node in the network of patients who live in high-income neighbourhoods. These results could help to personalize treatments, by allowing the identification of potential functioning areas to be prioritized in the treatment of FEP according to the patient's neighbourhood characteristic