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[EN] Perennial fruit production at the commercial scale, such as citrus fruits, begins with seedling production in a nursery. This stage lasts several months and involves different phases and the use of substrates and infrastructure. As the seedling does not produce fruit but does consume inputs, studying the environmental impacts associated with this stage becomes relevant, especially to understand its contribution to the total impact of the crop cycle. Despite the global relevance of fruit tree seedlings production, LCA studies in the literature focus on horticultural crop nurseries, and those on perennial tree nurseries do not consider both substrate and structures in the analysis, which is key for this type of crop since the main production system is soilless production in greenhouses. Thus, the main goal of this study is to quantify the environmental impacts related to the production of citrus fruit tree seedlings using LCA, analyse the main production system applied nowadays, and study its relevance with respect to the crop cycle. To this end, a certified Uruguayan citrus nursery was analysed, from which primary data was obtained. As well, methodological issues concerning water consumption and modelling emissions from input applications in soilless greenhouse systems are tackled. Results show that the main hotspots of the nursery stage are infrastructure production and peat transportation, which highlights the relevance of their inclusion when modelling the system. Extending the lifespan of the galvanised steel structures and decreasing substrate transport distances are shown to be effective measures to reduce environmental impacts. The contribution of the nursery stage to the citrus production cycle is negligible for almost all the impact categories assessed except cancer human toxicity, as it accounts for 0-3.6% of the impacts depending on the impact category. Great differences (from 10 to 400 times higher results on average) are observed when comparing the

The warp is a well-known undulation of the Milky Way disc. Its structure has been widely studied, but only since Gaia DR2 has it been possible to reveal its kinematic signature beyond the solar neighbourhood. In this work, we present an analysis of the warp traced by Classical Cepheids by means of a Fourier decomposition of their height (Z) and, for the first time, of their vertical velocity (Vz). We find a clear but complex signal that in both variables reveals an asymmetrical warp. In Z, we find the warp to be almost symmetric in amplitude at the disc’s outskirts, with the two extremes never being diametrically opposed at any radius and the line of nodes presenting a twist in the direction of stellar rotation for R > 11 kpc. For Vz, in addition to the usual m = 1 mode, an m = 2 mode is needed to represent the kinematic signal of the warp, reflecting its azimuthal asymmetry. The line of maximum vertical velocity is similarly twisted as the line of nodes and trails behind by ≈25°. We develop a new formalism to derive the pattern speed and change in amplitude with time A of each Fourier mode at each radius, via a joint analysis of the Fourier decomposition in Z and Vz. By applying it to the Cepheids we find, for the m = 1 mode, a constant pattern speed in the direction of stellar rotation of 9.2 ± 3.1 km s−1 kpc−1, a negligible A up to R ≈ 14 kpc and a slight increase at larger radii, in agreement with previous works.

[SPA] En los últimos años la incidencia de enfermedades no transmisibles, como la obesidad, han adquirido grado de pandemia global, así como otras patologías relacionadas como la diabetes tipo 2 o la enfermedad cardiovascular, que también están en aumento en muchos países. Una de las principales causas es la alimentación no equilibrada y pobre en frutas y hortalizas, que se une a un estilo de vida acelerado y sedentario, en el que el tiempo dedicado a la preparación de alimentos es escaso. En este contexto, resulta de interés el desarrollo de productos hortofrutícolas listos para el consumo, que permitan incorporar mayor cantidad de hortalizas a la dieta cotidiana. Las investigaciones del impacto que las tecnologías de procesado y conservación tienen sobre las propiedades nutricionales y bioactivas de los alimentos son cada vez más necesarias. El brócoli (Brassica oleracea var. italica) es una hortaliza versátil y altamente valorada por su importante contenido de fitoquímicos promotores de la salud, como glucosinolatos y ácidos hidróxicinámicos. Sin embargo, el brócoli tiene una vida útil corta, de pocos días, y el contenido de fitoquímicos disminuye drásticamente durante el almacenamiento si no se aplican condiciones adecuadas para su conservación. El envasado en atmósfera modificada (Modified Atmosfere Packaging - MAP) es una tecnología de conservación ampliamente utilizada en productos hortofrutícolas frescos. La misma consiste en envasar dichos productos en materiales plásticos que actúan como barrera a la difusión de gases, modificando el ambiente gaseoso que rodea al alimento, favoreciendo la extensión de su vida útil. Si bien existen estudios donde se ha evaluado el efecto del MAP en el contenido de compuestos bioactivos del brócoli, los resultados son contradictorios y las condiciones de conservación ensayadas son poco representativas de lo que ocurre en la cadena de suministro. Por otra parte, existe una urgente necesidad por encontrar nuevos envases que ga, [ENG] In recent years, the incidence of non-communicable diseases, such as obesity, has reached the level of a global pandemic, as well as other related conditions such as type 2 diabetes or cardiovascular disease, which are also on the rise in many countries. One of the main causes is an unbalanced and poor diet lacking in fruits and vegetables, combined with a fast-paced and sedentary lifestyle, where there is little time dedicated to food preparation. In this context, the development of ready-to-eat horticultural products becomes of interest, as they allow for the incorporation of more servings of vegetables into the daily diet. Research on the impact that processing and preservation technologies have on the nutritional and bioactive properties of foods is increasingly necessary. Broccoli (Brassica oleracea var. italica) is a versatile vegetable highly valued for its significant content of health-promoting phytochemicals, such as glucosinolates and hydroxycinnamic acids. However, broccoli has a short shelf life, lasting only a few days, and the phytochemical content decreases drastically during storage if appropriate measures for preservation are not taken. Modified Atmosphere Packaging (MAP) is a widely used preservation technology for fresh horticultural products. It involves packaging these products in plastic materials that act as barriers to gas diffusion, thereby modifying the gaseous environment surrounding the food and extending its shelf life. While there are studies evaluating the effect of MAP on the content of bioactive compounds in broccoli, the results are contradictory, and the conservation conditions tested are not fully representative of what occurs in the supply chain. Furthermore, there is an urgent need to find new packaging solutions that ensure the safety and quality of fruits and vegetables while concurrently reducing environmental impact. The objective of this Doctoral Thesis was to study the impact of different packaging films and storage

NASA sent the Double Asteroid Redirection Test (DART) mission to impact Dimorphos, the satellite of the asteroid binary system (65803) Didymos. DART will release LICIACube prior to impact to obtain high-resolution post-impact images. The impact will produce a crater and a large amount of material ejected at high speed (several tens of m s−1), producing an ejecta cone that will quickly disperse. We analysed an additional effect: the lofting of material at low velocity due to the generation of seismic waves that propagate inside Dimorphos, producing surface shaking far from the impact point. We divide the process into different stages: from the generation of impact-induced waves, the interaction of them with surface particles, the ejection of dust particles at velocities, and the prediction of the observability of the dust coma and trail. We anticipate the following observable effects: (i) generation of a dust cloud that will produce a hazy appearance of Dimorphos’ surface, detectable by LICIACube; (ii) brightness increase of the binary system due to enhancement of the cross-section produced by the dust cloud; (iii) generation of a dust trail, similar to those observed in some Active Asteroids, which can last for several weeks after impact. Numerical prediction of the detectability of these effects depends on the amount and size distribution of ejected particles, which are largely unknown. In case these effects are observable, an inversion method can be applied to compute the amount of ejected material and its velocity distribution, and discuss the relevance of the shaking process. © 2022 The Author(s) Published by Oxford University Press on behalf of Royal Astronomical Society.

Coastal lagoons are among the most productive and biodiverse systems in the world and are important sentinels of climate change. The Mar Menor is one of the largest coastal lagoons in the Mediterranean, providing a variety of ecosystem services and resources to the community. However, in recent decades this lagoon has suffered drastic changes and degradation caused by human activities. We analyzed the concentration of dissolved organic carbon (DOC) and the optical properties of dissolved organic matter (DOM) in the water column and sediment pore water during the summer and winter of 2018 and during eighteen months from 2016 to 2018. Overall, we found that the composition of DOM is mainly related to and enhanced by anthropogenic activities and microbial metabolism. DOM enters the lagoon via urban and agricultural runoff, drainage systems, and wastewater treatment plants. Additionally, strong microbial metabolism in sediments leads to differences in DOM composition between water and sediments. In the water column, humic-like components accounted for 71 % of the total DOM, while protein-like compounds were most abundant in sediment pore water. We observed a strong seasonal variability associated with precipitation and the system collapse in 2016 (phytoplankton bloom), which resulted in the death of 80 % of macrophytes. The sediments act as a source of DOM to the overlying water, likely due to relatively high organic matter content and intense microbial activity, primarily through anaerobic pathways. Benthic fluxes of DOC ranged from 5.24 to 33.30 mmol m−2 d−1, being higher in winter than summer 2018 and decreasing from north to south, likely related to lower residence time in the northern basin, groundwater discharge and accumulation of organic matter from the dead meadows. We estimate a net flux of DOC from the Mar Menor toward the Mediterranean Sea of 1.57 × 107 mol yr−1

Glioblastoma (GBM), as the most central nervous system (CNS) intractable disease, has spoiled millions of lives due to its high mortality. Even though several efforts have been made, the existing treatments have had limited success. In this sense, we studied a lead compound, the boron-rich selective epidermal growth factor receptor (EGFR)-inhibitor hybrid 1, as a potential drug for GBM treatment. For this end, we analyzed the in vitro activity of hybrid 1 in a glioma/primary astrocytes coculture, studying cellular death types triggered by treatment with this compound and its cellular localizations. Additionally, hybrid 1 concentrated boron in glioma cells selectively and more effectively than the boron neutron capture therapy (BNCT)-clinical agent 10B-l-boronophenylalanine and thus displayed a better in vitro-BNCT effect. This encouraged us to analyze hybrid 1 in vivo. Therefore, immunosuppressed mice bearing U87 MG human GBM were treated with both 1 and 1 encapsulated in a modified liposome (recognized by brain-blood barrier peptide transporters), and we observed a potent in vivo per se antitumor activity (tumor size decrease and animal survival increase). These data demonstrate that 1 could be a promising new targeted therapy for GBM.

Recreational spearfishing is a fishing method that occurs globally, yet receives considerably less attention in the scientific literature relative to other recreational fishing methods, such as angling. Lack of scientific information on spearfishing may negatively affect the development and management of marine recreational fisheries. We conducted a systematic review of 102 peer-reviewed papers published between 1967 and 2022 pertaining to marine recreational spearfishing. Based on this literature review, we provide an overview of key insights across social, economic, and ecological dimensions of marine recreational spearfishing. While spearfishers represent less than 5% of marine recreational fishers, the participants are younger and may differ from recreational anglers in their motivations, with suggestions of increased well-being generated from a close connection with the sea during underwater fishing. Recreational spearfishers mostly target species of moderate to high levels of vulnerability that are mid to high trophic level carnivores. Though spearfishers can deliberately target larger individuals of exploited populations, this is not a generalizable pattern. Despite a growing body of research on the ecological impacts of marine recreational spearfishing, there is limited knowledge of these effects and their mechanisms across biological levels of organization (e.g., individual, population, community and ecosystem) compared with those of other fishing methods. Recreational spearfishers can contribute to advances in marine ecological knowledge, and inclusive participatory management could represent a key step towards transformative sustainable development of marine recreational spearfishing. Throughout the review, we identify gaps in the research and areas where future research is needed to better inform the socio-economic importance, ecosystem impacts and future management of marine recreational spearfishing

Additional file 1: QIIME2 pipeline used to process and combine the two 16S sequence datasets.

Additional file 2: Fig. S1. Evolution of heritability estimates along iterations in an holobiont model in CRP to illustrate convergence of the MCMC chain. Fig. S2. PCA for the individual batches and the merged dataset of CLR-transformed ASV abundances. Fig. S3. Distribution of reads per sample in the merged 16S sequencing data after quality control. Fig. S4. Distribution of CLR-transformed ASV abundances, averaged across samples. Fig. S5. Distribution of the ASV heritability estimates.

Additional file 3: Table S1. Correlation of the Euclidean distances between samples in each pair of datasets. Table S2. ASV responsible for 30% of the estimated b2 under the best predictive model for LYM_PHAGO_FITC.

We analyzed, from a cultural astronomy perspective, the relationship between the orientations of five mound sites and different astronomical events in the lowland region of Uruguay. We found significant relationships between the orientations of the mounds and the Southern Cross/Milky Way and the full Moon during the winter solstice ca. 3000 years BP. These relationships, meanings and senses to different native peoples of South America were explored from the literature of travelers’ and naturalists’ chronicles, alongside the ethnohistorical, ethnographic and archaeological literature. In particular, we highlighted the link among those peoples of the area of the Southern Cross/ Milky Way with a mythical Ñandú (Rhea americana). Such an interpretation has allowed us to raise the possibility that we are being faced with the integration of knowledge of the sky in the form of the social construction of inhabited space and the configuration of the landscape.

This study aimed to evaluate the nutritional value and potential health claims for reduced sugar biscuits containing extruded brewers’ spent grain (EBSG) and fructooligosaccharides (FOS). One traditional biscuit with added sugar and three reduced sugar biscuits containing 15.2 % FOS and EBSG (0, 8 and 17 %), with nutrition claims “high in fiber” and “source of protein” for those containing 17 % of EBSG, were formulated. Biscuits were characterized by analysis of nutrients and bioactive compounds before and after digestion under in vitro enzymatic oral-gastro-intestinal and colonic fermenting conditions. The bioaccessibility of antioxidants, anti-inflammatory and antidiabetic compounds in biscuits’ intestinal digests and short-chain fatty acids in colonic samples was analyzed. EBSG-added biscuits showed significantly lower (p < 0.05) glucose intestinal bioaccessibility and significantly higher (p < 0.05) phenolic compounds intestinal bioaccessibility compared to biscuits without EBSG. EBSG-added biscuits showed significantly (p < 0.05) higher in vitro antidiabetic potential than the other did. Moreover, the intestinal digest of biscuits containing 17 % EBSG exhibited significantly (p < 0.05) better in vitro inhibition of intracellular ROS formation and in vitro anti-inflammatory properties. FOS addition (p < 0.05) significantly improved the production of butyric acid while EBSG did for valeric acid which possess chemoprotective effect. In conclusion, the combined use of FOS (15.2 %) and EBSG (17 %) allowed obtaining a human healthier snack formulation for satisfying consumers’ demands and achieving nutrition security.

En este texto se parte de una contextualización de la problemática de los cerritos de indios en el marco de las políticas patrimoniales en Uruguay. A continuación, se describe someramente un proyecto de gestión patrimonial y producción responsable como bases para el desarrollo social, llevado a cabo en un conjunto de cerritos de India Muerta (Rocha, Uruguay), entre 2016 y 2021. En la parte subsiguiente se caracteriza el concepto clásico de “cadena de valor del patrimonio cultural”, y se argumenta la necesidad de una actualización del mismo. A partir de la experiencia del proyecto en Uruguay, se realiza una reflexión teórico-metodológica orientada a dicha actualización. Se pone de manifiesto la necesidad de un replanteamiento de las bases ontológicas de la gestión del paisaje y el patrimonio, así como una ampliación y relativización del rol de la ciencia (en concreto, de la arqueología), en el marco de un proceso de patrimonialización en el que diversas dimensiones valorativas deben ser tenidas en cuenta. Esta actualización implica el paso de un patrimonio entendido como objeto o conjunto de objetos a valorizar, a un patrimonio que funciona como medio para la transformación e innovación social.

Broccoli heads are typically packaged and marketed in petroleum-based heat-shrinkable or stretch-cling films that are not biodegradables. These types of films are not compatible with global strategies to reduce plastics waste. One of these strategies is development of biodegradable packaging materials that can be used as an alternative food package. Therefore, a biodegradable stretch-cling film, based on poly(butylene-adipate-co-terephthalate) and poly(lactide) blend (Nature Fresh®), for its application in broccoli head packaging has been studied. To determine suitability of Nature Fresh® film for broccoli packaging, broccoli heads were wrapped in this film and stored at 2°C for 35 d. Broccolis without packaging and wrapped in linear low density polyethylene (LLDPE) were used as controls. Physicochemical and sensory evaluation were determined during storage to study the effect of films on quality and shelf life of broccoli. On the one hand, broccolis packaged in Nature Fresh® and LLDPE presented similar evolution in internal atmosphere composition, color, hydroxycinnamic acids and carotenoids content and overall appearance. On the other hand, weight loss and decrease in firmness were significantly higher in broccoli wrapped in Nature Fresh® compared to LLDPE, but this had no impact on overall appearance. Nature Fresh® extended postharvest storage of broccoli heads at least 21 d at 2ºC, which is an adequate time to market. In conclusion, Nature Fresh® film was a viable packaging alternative to LLDPE to preserve broccoli quality during postharvest storage, while fulfilling global strategies to reduce plastics waste.

Olive leaves (OL) are considered a potential source of bioactive compounds mainly due to its high content of phenolic compounds, widely known as natural antioxidants. The main objective of this study was to compare the performance of three OL extracts obtained by different extraction techniques in protecting canola oil against oxidative damage. The technologies evaluated were maceration with ethanol/water 75:25 (v/v), supercritical fluid extraction with CO2 (SC–CO2) and SC-CO2 with 10% ethanol as modifier (SC–CO2/EtOH). Each extract was analyzed as for total phenolic compounds (TPC), antioxidant activity (ABTS assay) and phenolic composition by reversed phase liquid chromatography-quadrupole-time of flight mass spectrometry. The oxidative stability of canola oil with or without the incorporation of 250 mg/kg of each extract was assessed during five weeks of storage at 60 °C. Peroxide, K232, K270, and Rancimat values, besides tocopherols content were determined. Macerated extract showed the highest TPC and antioxidant activity, but both SC-CO2 extracts were more effective in preserving tocopherols. In addition, SC-CO2 extracts delayed the oxidation progress as they lead to higher induction periods than control and macerated extracts, and a slower increase in peroxide values. Results obtained reinforce the use of supercritical fluid technology to obtain antioxidants compounds from natural sources.

Alpeorujo is a semi-solid residue from the olive oil industry that accounts for around 80–85% of the total processed olives; it contains phenolic compounds that could be used as natural preservatives in the food industry. The objective of this study was to optimize supercritical fluid extraction (SFE) of antioxidant compounds from alperujo using a Box-Behnken Design and response surface methodology. Antioxidant (ABTS and ORAC assays) and antimicrobial (MIC) potential of SFE extracts from Arbequina and Coratina were compared for extracts obtained in the optimal conditions to maximize the antioxidant activity (200 bar, 60 °C and 10% ethanol as modifier). Extracts from Coratina presented a significantly higher (p < 0.05) TPC (1487–2073 mg GAE/kg) and tocopherol content (345–454 ppm), although, in general, a correlation between these values and antioxidant activity of extracts was not observed. Moreover, Arbequina and Coratina SFE extracts showed moderate bacterial inhibitory potential against most of the bacteria studied.

This paper presents new data on the spatial organization of mound-builder groups in the India Muerta wetlands, Uruguay. This area presents the beginning of land architecture in the region (ca. 4800–5000 years BP), associated with more arid climate. This construction tradition continues and intensifies, mainly from ca 3000 years BP, from the establishment of warmer and damper conditions. New sources of information and geospatial technologies have made it possible to locate mound sites with greater precision, as well as to analyze settlement patterns. Indigenous communities occupied areas of hills, plains and wetlands, showing differences but also regularities in spatial organization in each area. In the whole area, earthen mound complexes form groups of different orders, from regional to domestic units, configured by mounds, negative structures and limited spaces. The location of the mounds is primarily in dry areas, known locally as islands, which are prominent in the landscape during floods in this wetland-dominated environment. Through this analysis of the landscape, this work delves into the underlying logic of the social construction of the territory. The results achieved in this paper are consistent with previous research suggesting planned occupation associated with villages integrated within broader regional systems.

This study aimed to determine the effect of the extrusion process on the nutritional and bioactive profiles of brewer’s spent grain (BSG), contributing to nutrition security by applying a circular economy concept. Response surface methodology was used to optimize the effect extrusion parameters (moisture content, screw speed, and barrel temperature ) had on BSG’s soluble dietary fiber, free glucose, and overall antioxidant capacity. Proximate composition analyses, amino acid profile, extractable polyphenolic content, and antioxidant capacity of BSG and brewer’s spent grain extruded under optimal conditions (BSGE) were carried out. Food safety was analyzed by their microbiological quality, gluten, and acrylamide content. Optimal extrusion conditions were 15.8% of moisture content, 164.3 revolutions per min and 122.5 °C. BSGE presented 61% more soluble dietary fiber than BSG, lower digestible starch, 0.546% of free glucose, and protein quality parameters mostly like those reported for egg, soy, and milk. Despite this, BSG’s overall antioxidant capacity was not improved after thermomechanical processing; BSGE had significantly higher extractable polyphenolic content in its alkali extracts, which were determined qualitatively by high-performance liquid chromatography quadrupole time-of-flight assay in its hydro-alcoholic acid extracts. Furthermore, although it is not gluten free, BSGE is a safe food ingredient with acceptable microbiological quality and no acrylamide.

This study aimed to evaluate the effect of the extrusion process on the bioaccessibility of brewers’ spent grain (BSG) nutrients (carbohydrates and proteins) and non-nutrients (bioactive compounds). BSG and extruded BSG (EBSG) were digested in vitro simulating human oral-gastro-intestinal digestion and colonic fermentation. The duodenal bioaccessibility of glucose, amino acids and phenolic compounds was analyzed. The fermentability of the dietary fiber was assessed by analysis of short-chain fatty acids. Additionally, assessment of the bioaccessibility of phenolic compounds after colonic fermentation was undertaken. The antioxidant, anti-inflammatory and antidiabetic properties of the bioaccessible compounds were studied. Extrusion caused no change in the digestibility of gluten and glucose bioaccessibility (p > 0.05). Moreover, the bioaccessibility of amino acids and phenolic compounds significantly increased (p < 0.05) due to extrusion. However, higher short-chain fatty acid content was formed in colonic fermentation of BSG (p < 0.05) compared to EBSG. The latter inhibited intracellular ROS formation in IEC-6 cells and showed anti-inflammatory properties in RAW264.7 cells. With respect to antidiabetic properties, glucose absorption was lower, and the inhibition of carbohydrases higher (p < 0.05), in the presence of EBSG compared to BSG. The effects of EBSG and BSG digests on glucose transporters were not significantly different (p > 0.05). In conclusion, extrusion positively affected the nutritional value and health-promoting properties of BSG.

Cooking vegetables in microwave bags is becoming a popular domestic cooking method, being relevant to know how this cooking method affects health-promoting phytochemicals of staples such as broccoli. The aim of this work was to study the effect of microwave bag cooking versus conventional microwaving on bioactive compound content (glucosinolates and hydroxycinnamic acid derivatives) and other quality parameters (such as antioxidant capacity, mineral content and microbial load) of broccoli florets. The influence of cooking time on bioactive compounds content was also evaluated. The study was carried out in two independent experiments; using intact broccoli and broccoli preprocessed in industry. Microwave bag cooked broccoli for 5 min (following label recommendation) showed higher glucosinolate content retention compared to conventional microwaving. Results suggest that volatilization could be an important phenomenon in reduction of glucosinolates during microwave cooking of broccoli florets. Glucosinolate profile did not change after cooking, regardless of cooking method applied. Furthermore, microwave bag cooked broccoli presented higher antioxidant capacity (by DPPH assay) than conventional microwaved broccoli. Hydroxycinnamic acid derivatives content was reduced in microwave cooking, regardless of method applied. Altogether, the use of microwave bags for microwaving is a novel method that retains main bioactive components of broccoli. This option is a fast, easy and considerably clean cooking option to fulfill modern consumer needs.

Gabon buck is a breed with little marked seasonality in our latitude (Uruguay, 35° SL). The role of thyroid hormones on the regulation of their seasonal reproductive activity and sperm cryoresistance is unknown. Seasonal changes in testosterone concentration can affect sperm variables, but the influence of testosterone changes on sperm cryoresistance in other species determines that the recommended time for freezing sperm does not coincide with the period with greater sperm fresh quality. The objectives of the present work were to (i) describe the thyroxine seasonal pattern in bucks in a subtropical area, and its association with annual changes in sperm variables; (ii) relate the seasonal changes of testosterone and thyroxine concentrations with the sperm cryoresistance. For one year, semen of 10 adult Gabon bucks was collected by electroejaculation every two weeks. After sperm selection, the sample was frozen. Testosterone and thyroxine concentrations varied according to the month (P < 0.0001). Testosterone reached the greatest values in April (P < 0.0001) and May (P < 0.0001) and thyroxine reached minimum values (P < 0.0001) in the same months. During these months, a negative correlation ratio (CR) was found between testosterone concentration and CR-functional membrane (R = - 0.50; P < 0.0001). CR values for most sperm variables decreased during March-May, coinciding with the presence of maximum testosterone concentrations. In conclusion, high testosterone levels are associated with the worst sperm response to freezing-thawing process. Thyroxine concentrations have a strong seasonal pattern, but there was no relationship to sperm cryoresistance.

Seafloor structures related to the emission of different fluids, such as submarine mud volcanoes (MVs), have been recently reported to largely contribute with dissolved organic matter (DOM) into the oceans. Submarine MVs are common structures in the Gulf of Cádiz. However, little is known about the biogeochemical processes that occur in these peculiar environments, especially those involving DOM. Here, we report DOM characterization in the sediment pore water of three MVs of the Gulf of Cádiz. Estimated benthic fluxes of dissolved organic carbon (DOC) and chromophoric DOM (CDOM) were higher than in other marine sediments with an average of 0.11 ± 0.04 mmol m−2 d−1 for DOC and ranging between 0.11 and 2.86 m−1 L m−2 d−1, for CDOM. Protein-like components represented ~ 70% of the total fluorescent DOM (FDOM). We found that deep fluids migration from MVs (cold seeps) and anaerobic production via sulfate-reducing bacteria represent a source of DOC and FDOM to the overlying water column. Our results also indicate that fluorescent components can have many diverse sources not captured by common classifications. Overall, MVs act as a source of DOC, CDOM, and FDOM to the deep waters of the Gulf of Cádiz, providing energy to the microbial communities living there

Citrus fruit are sensitive to chilling injury (CI) during cold storage, a peel disorder that causes economic losses. C-repeat binding factors (CBFs) are related to cold acclimation and tolerance in different plants. To explore the role of Citrus CBFs in fruit response to cold, an in silico study was performed, revealing three genes (CBF1, CBF2, and CBF3) whose expression in CI sensitive and tolerant cultivars was followed. Major changes occurred at the early stages of cold exposure (1–5 d). Interestingly, CBF1 was the most stimulated gene in the peel of CI-tolerant cultivars (Lisbon lemon, Star Ruby grapefruit, and Navelina orange), remaining unaltered in sensitive cultivars (Meyer lemon, Marsh grapefruit, and Salustiana orange). Results suggest a positive association of CBF1 expression with cold tolerance in Citrus cultivars (except for mandarins), whereas the expression of CBF2 or CBF3 genes did not reveal a clear relationship with the susceptibility to CI. Light avoidance during fruit growth reduced postharvest CI in most sensitive cultivars, associated with a rapid and transient enhance in the expression of the three CBFs. Results suggest that CBFs-dependent pathways mediate at least part of the cold tolerance responses in sensitive Citrus, indicating that CBF1 participates in the natural tolerance to CI.

[Aim] Global anthropogenic changes have altered biogeography and phenology of marine populations, thereby promoting a spatial reconfiguration in the functioning of marine ecosystems. Among these changes, massive proliferations of jellyfish in temperate latitudes warn of potential alterations in biogeochemical fluxes, ecosystems’ structure and assets, and the services they provide to human welfare. Understanding driving factors shaping large-scale patterns of jellyfish proliferations is a pressing need in global ecology and sustainability science. Using a comprehensive dataset of the largest blooming scyphomedusae in southern European seas we test, over broad space–time scales, current hypotheses relating jellyfish dynamics to warming and eutrophication., [Location] Southern European seas., [Time period] 1875–2019., [Major taxa studied] Rhizostoma pulmo., [Methods] We have gathered historical and contemporaneous records of R. pulmo from the Mediterranean and Black Seas over the last two centuries (7,359 records). Generalized statistical models were used to assess the influence of thermal (latitudinal) and productivity (longitudinal) gradients on the biogeographical patterns, and the species’ phenology at large and regional scales., [Results] Rhizostoma pulmo abundance exhibited an enhanced magnitude and frequency in recent decades, concurrently with positive temperature anomalies. We found that the latitudinal temperature gradient, but not productivity, shaped long-term bloom intensity and biogeographical patterns of the species. Our analysis further uncovered a significant effect of the interannual variability of spring temperature on the species’ phenology over the period 2008–2018, with warmer springs favouring an earlier start (c. 3 months) and a longer duration (from 5 to 7 months) of jellyfish season., [Main conclusions] Among the current hypotheses linking jellyfish changes with anthropogenic disturbances, only the warming-based hypothesis gained support over wide space–time scales, while the eutrophication-based hypothesis mainly applied at local scales. Hence, biogeographical patterns of R. pulmo are shaped by the latitudinal temperature gradient, while the species bloom dynamics echo variations in ecoregion thermal regimes.

The reduction of the environmental impact represents an important challenge for the winemaking industry. Red grape by-products are composed of several health-promoting components such as polyphenols and dietary fiber, which may be employed as sustainable food ingredients. Although the bioactive properties of red grape pomace, one of the most abundant byproducts of the winemaking industry worldwide, have been extensively documented by in vitro studies, there is a lack of information on the bioaccessibility, bioavailability and food applications that will subsequently affect the contribution of grape byproduct bioactive compounds to these properties. Therefore, the present chapter aimed to present some of the latest grape byproducts in vitro bioactive properties, as well as novel and updated knowledge related to its bioaccessibility, bioavailability and food applications.

Although the soil bacterium Pseudomonas putida KT2440 bears a bona fide adenylate cyclase gene (cyaA), intracellular concentrations of 3′,5′-cyclic adenosine monophosphate (cAMP) are barely detectable. By using reporter technology and direct quantification of cAMP under various conditions, we show that such low levels of the molecule stem from the stringent regulation of its synthesis, efflux and degradation. Poor production of cAMP was the result of inefficient translation of cyaA mRNA. Moreover, deletion of the cAMP-phosphodiesterase pde gene led to intracellular accumulation of the cyclic nucleotide, exposing an additional cause of cAMP drain in vivo. But even such low levels of the signal sustained activation of promoters dependent on the cAMP-receptor protein (CRP). Genetic and biochemical evidence indicated that the phenomenon ultimately rose from the unusual binding parameters of cAMP to CRP. This included an ultratight cAMP-Crp affinity (K of 45.0 ± 3.4 nM) and an atypical 1:1 effector/dimer stoichiometry that obeyed an infrequent anti-cooperative binding mechanism. It thus seems that keeping the same regulatory parts and their relational logic but changing the interaction parameters enables genetic devices to take over entirely different domains of the functional landscape.

Biocatalytic cascades afford the development of economically sustainable and green processes. Herein we examined the unprecedented coupling of co-immobilised Gluconobacter oxydans and an isolated transaminase to synthesise serinol from glycerol. Through this approach, we manufactured up to 36 mM serinol, the highest titer ever reported for a non-fermentative biosynthesis. More importantly, similar productivities are obtained starting from the industrial by-product crude glycerol, demonstrating the possibilities of this hybrid heterogenenous biocatalyst for valorising bio-based raw materials.

In most ram breeds, there are annual changes in testosterone serum concentration and semen quality. This study aimed to determine if eCG administration to rams during the non-breeding season improves the reproductive activity. In the first study, we compared the effects of administering three doses of eCG (0, 400 or 700 IU). In the second experiment we compared the effects of administering eCG to rams from two breeds originated in different latitudes, and thus, with different expected reproductive seasonal pattern (Highlander; HL and Texel; TEX). The first study was performed during the mid non-breeding season with three groups of rams: treated with 400 IU (group eCG400) or 700 IU of eCG (group eCG700), or untreated controls (group CON). Treated rams received three doses of eCG administered every 6 days. Testosterone concentration, testicular traits (scrotal circumference, testicular and sexual skin flush, and ultrasonographic testes pixel color intensity) and seminal characteristics were determined from five days before the first eCG administration until Day 24. Testosterone concentration was greater in eCG700 than in eCG400 (P <0.001) and CON (P <0.0001) rams, and in eCG400 than in CON rams (P = 0.0002). However, the treatment did not modify the testicular traits and seminal quality. The second study was performed with 15 HL and 17 TEX rams; while 8 HL and 9 TEX rams received two doses of 1000 IU of eCG (Days 0 and 5), the other rams remained as untreated controls. Scrotal circumferences, seminal traits (fresh and thawed semen, and cryopreservation ratio –CR-) as well as the sexual behavior were determined, twice before eCG administration and during 3 weeks. The treatment did not modify the scrotal circumference nor the fresh sperm traits evaluated. However, the CR of motile and progressive motile sperm were greater in eCG treated rams than in control rams on Day 7 (P = 0.01 for both). Treated rams had their second and third ejaculations earlier than control rams

About 95 % of people diagnosed with glioblastoma die within five years. Glioblastoma is the most aggressive central nervous system tumour. It is necessary to make progress in the glioblastoma treatment so that advanced chemotherapy drugs or radiation therapy or, ideally, two‐in‐one hybrid systems should be implemented. Tyrosine kinase receptors–inhibitors and boron neutron capture therapy (BNCT), together, could provide a therapeutic strategy. In this work, sunitinib decorated‐carborane hybrids were prepared and biologically evaluated identifying excellent antitumoral‐ and BNCT‐agents. One of the selected hybrids was studied against glioma‐cells and found to be 4 times more cytotoxic than sunitinib and 1.7 times more effective than 10B‐boronophenylalanine fructose complex when the cells were irradiated with neutrons.

Efficient engagement with the envelope glycoprotein membrane-proximal external region (MPER) results in robust blocking of viral infection by a class of broadly neutralizing antibodies (bnAbs) against human immunodeficiency virus (HIV). Developing an accommodation surface that engages with the viral lipid envelope appears to correlate with the neutralizing potency displayed by these bnAbs. The nature of the interactions established between the antibody and the lipid is nonetheless a matter of debate, with some authors arguing that anti-MPER specificity arises only under pathological conditions in autoantibodies endowed with stereospecific binding sites for phospholipids. However, bnAb-lipid interactions are often studied in systems that do not fully preserve the biophysical properties of lipid bilayers, and therefore, questions on binding specificity and the effect of collective membrane properties on the interaction are still open. Here, to evaluate the specificity of lipid interactions of an anti-MPER bnAb (4E10) in an intact membrane context, we determine quantitatively its association with lipid bilayers by means of scanning fluorescence correlation spectroscopy and all-atom molecular dynamic simulations. Our data support that 4E10 establishes electrostatic and hydrophobic interactions with the viral membrane surface and that the collective physical properties of the lipid bilayer influence 4E10 dynamics therein. We conclude that establishment of peripheral, nonspecific electrostatic interactions with the viral membrane through accommodation surfaces may assist high-affinity binding of HIV-1 MPER epitope at membrane interfaces. These findings highlight the importance of considering antibody-lipid interactions in the design of antibody-based anti-HIV strategies.

The Gulf of Cádiz, located in the Southern Iberian Atlantic Basin, connects the North Atlantic Ocean and the Mediterranean Sea and receives freshwater input from three main estuaries: Guadalquivir, Guadiana, and Tinto-Odiel. These estuaries differ in their hydrology, basin characteristic, and land use. One of them, Tinto-Odiel, is one of the most polluted estuaries in the world. However, little is known about the export of DOM from these estuaries to the Gulf of Cádiz. In this work, the estuaries were sampled during the dry season. Dissolved organic matter (DOM) was chemically characterized along a longitudinal gradient to better understand the reactivity and fate of carbon exported to the Gulf of Cádiz. We also performed a tidal study at the mouth of each estuary to understand how the tide affects the composition of DOM and its export to the coastal zone. Fluorescent DOM (FDOM) modeling employing multivariate parallel factor analysis and pyrolysis-gas chromatography-mass spectroscopy analysis (Py-GC–MS) were used to characterize DOM. DOM from Guadalquivir and Guadiana estuaries presented a predominant allochthonous origin with humic-like compounds making up to ~80% of the total fluorescent DOM. These estuaries receive lateral inputs from surrounding watersheds and agricultural practices. Instead, in Tinto-Odiel estuary, DOM was predominantly autochthonous with a higher content of protein-like material associated with the prevalence of the marine influence over the low water discharges and anthropogenic pollution from industrial activities. Tidal cycles affected the distribution of DOM and its quality with higher humic-like material during low tide and protein-like substances predominating during high tide. During the dry season, the three estuaries represented a source of DOM to the Gulf of Cádiz. Guadalquivir was the main contributor with 1.25 Kg C s and 0.22 × 10 m s, for DOC and CDOM, respectively. Around 70% of the FDOM exported from the three estuaries was of

The antimicrobial properties of silver nanoparticles (AgNPs) have made them ubiquitous in a number of real-world industrial applications; however, the antimicrobial mode of action of biogenic AgNPs is not entirely understood. The use of Raman spectroscopy can provide molecular fingerprint information on various chemical and biochemical components in complex systems like microbial cultures, without the need for any complex sample pre-treatment. Consequently, the antimicrobial mechanism of AgNPs can be inferred through morphological and compositional changes of microbial cells that are monitored via changes in Raman band profiles. Here we show the synthesis of biogenic AgNPs using the extracellular cell-free filtrates of Penicillium expansum. The antimicrobial activity of the Penicillium expansum synthesized silver nanoparticles (hereafter PeNPs) was evaluated and the interactions between the nanoparticles and Escherichia coli were studied using Transmission Electron Microscopy (TEM) and Environmental Scanning Electron Microscopy (ESEM), showing the attachment of PeNPs to the surface of the bacteria and rupture of the bacterial cell membrane. Importantly, we show how Confocal Raman Microscopy can be used as an innovative approach to study the antimicrobial mechanisms, the results of which confirm that the PeNPs induce damage to bacterial and fungal cells, resulting in critical changes to polysaccharides, lipids, proteins and nucleic acids.

This paper presents the levels and distribution of 210Po in different compartments of a coastal lagoon on the east coast of Uruguay (South America). Activity concentrations of 210Po have been obtained in different matrices, such as water, superficial sediments, clams (Diplodon sp.), freshwaters snails (Pomacea sp.), zooplankton, and fishes (Jenynsia sp.), collected at different points of the lagoon and during several sampling campaigns. In addition, the organic matter content of the sediment was determined to study the variation of 210Po along the lagoon. The activity concentrations of 210Po in the water samples are in the range between 1.1 ± 0.2–3.5 ± 0.4 mBq/L while in the sediment samples vary between 17.1 ± 1.4 and 540 ± 12 Bq/kg, DW. In the case of biota, the ranges obtained were 182 ± 5–265 ± 6 Bq/kg, DW in clams and 134 ± 4–1245 ± 16 Bq/kg, DW in snail samples. A good correlation of 210Po with the organic fractions of the sediment was observed (r = 0.8798, p-value < 0.001), being obtained high values for the distribution coefficient Kd (104 -105). In the biota samples, a clear difference was observed in the 210Po concentration values in both species, mainly due to the different feeding habits of both aquatic organisms, as it is reflected in the associated concentration ratios (CR). In this paper, a good set of results of 210Po, Kd, and CR have been obtained in different matrices, enhancing the limited archives available for modelers concerning these parameters for 210Po and freshwater systems.

Microorganisms offer an alternative green and scalable technology for the synthesis of value added products. Fungi secrete high quantities of bioactive substances, which play dual-functional roles as both reducing and stabilizing agents in the synthesis of colloidal metal nanoparticles such as silver nanoparticles, which display potent antimicrobial properties that can be harnessed for a number of industrial applications. The aim of this work was the production of silver nanoparticles using the extracellular cell free extracts of Phanerochaete chrysosporium, and to evaluate their activity as antimicrobial and antibiofilm agents. The 45–nm diameter silver nanoparticles synthesized using this methodology possessed a high negative surface charge close to −30 mV and showed colloidal stability from pH 3–9 and under conditions of high ionic strength ([NaCl] = 10–500 mM). A combination of environmental SEM, TEM, and confocal Raman microscopy was used to study the nanoparticle-E. coli interactions to gain a first insight into their antimicrobial mechanisms. Raman data demonstrate a significant decrease in the fatty acid content of E. coli cells, which suggests a loss of the cell membrane integrity after exposure to the PchNPs, which is also commensurate with ESEM and TEM images. Additionally, these biogenic PchNPs displayed biofilm disruption activity for the eradication of E. coli and C. albicans biofilms.

One of the driving forces of carcinogenesis in humans is the aberrant activation of receptors; consequently, one of the most promising mechanisms for cancer treatment is receptor inhibition by chemotherapy. Although a variety of cancers are initially susceptible to chemotherapy, they eventually develop multi-drug resistance. Anti-tumor agents overcoming resistance and acting through two or more ways offer greater therapeutic benefits over single-mechanism entities. In this study, we report on a new family of bifunctional compounds that, offering the possibility of dual action (drug + radiotherapy combinations), may result in significant clinical benefits. This new family of compounds combines two fragments: the drug fragment is a lapatinib group, which inhibits the tyrosine kinase receptor activity, and an icosahedral boron cluster used as agents for neutron capture therapy (BNCT). The developed compounds were evaluated in vitro against different tyrosine kinase receptors (TKRs)-expressing tumoral cells, and in vitro–BNCT experiments were performed for two of the most promising hybrids, 19 and 22. We identified hybrid 19 with excellent selectivity to inhibit cell proliferation and ability to induce necrosis/apoptosis of glioblastoma U87 MG cell line. Furthermore, derivative 22, bearing a water-solubility-enhancing moiety, showed moderate inhibition of cell proliferation in both U87 MG and colorectal HT-29 cell lines. Additionally, the HT-29 cells accumulated adequate levels of boron after hybrids 19 and 22 incubations rendering, and after neutron irradiation, higher BNCT-effects than BPA. The attractive profile of developed hybrids makes them interesting agents for combined therapy.

Antler growth is a costly yearly process supplied by daily nutrition and body reserves. Conditions for antler growth are rarely optimal. Thus, certain decrease in antler quality due to depletion of resources, known as physiological exhaustion, is frequently observed along the main beam. Testosterone promotes the mineralization in the last phase of antler growth, and its concentration change in male deer according to the socio-sexual environment: it is greater in single males housed only with females than in males housed only with other males (female effect). We hypothesised that the socio-sexual environment affect antler mineralization; especially in the top of the antler which is the last part mineralising and subjected to the described physiological exhaustion, but also the one mineralising when testosterone levels are higher. We tested this in antlers of pampas deer (Ozotoceros bezoarticus) from captive animals living in the two social contexts described. All the animals had an ad libitum high nutrition plan, so results are not due to nutritional differences. The antlers from males living with females suffered a lower physiological exhaustion (i.e., lower percent decrease in the top compared to the base of the antler) in the two most important minerals: Ca and P, but also similar tendencies in other bone-biology related trace elements like Sr and Li. Our results highlight that the socio-sexual environment, particularly living continuously in close contact with females, promotes antler mineralization probably through modifications in testosterone secretion.

Hybrid and composite nanoparticles represent an attractive material for enzyme integration due to possible synergic advantages of the structural builders in the properties of the nanobiocatalyst. In this study, we report the synthesis of a new stable hybrid nanobiocatalyst formed by biomimetic silica (Si) nanoparticles entrapping both Horseradish Peroxidase (HRP) (EC 1.11.1.7) and magnetic nanoparticles (MNPs). We have demonstrated that tailoring of the synthetic reagents and post immobilization treatments greatly impacted physical and biocatalytic properties such as an unprecedented ~280 times increase in the half-life time in thermal stability experiments. The optimized nanohybrid biocatalyst that showed superparamagnetic behaviour, was effective in the batch conversion of indole-3-acetic acid, a prodrug used in Direct Enzyme Prodrug Therapy (DEPT). Our system, that was not cytotoxic per se, showed enhanced cytotoxic activity in the presence of the prodrug towards HCT-116, a colorectal cancer cell line. The strategy developed proved to be effective in obtaining a stabilized nanobiocatalyst combining three different organic/inorganic materials with potential in DEPT and other biotechnological applications.

Cancer is the second leading cause of death worldwide. Herein, a strategy to quickly and efficiently identify novel lead compounds to develop anticancer agents, using green multicomponent reactions followed by antiproliferative activity and structure–activity relationship studies, is described. A second-generation focused library of nitric oxide-releasing compounds was prepared by microwave-assisted Passerini and Ugi reactions. Nearly all compounds displayed potent antiproliferative activities against a panel of human solid tumor cell lines, with 1-phenyl-1-[(tert-butylamino)carbonyl]methyl 3-[(3-phenylsulfonyl-[1,2,5]oxadiazol-4-yl N-oxide)oxy]benzoate (4 k) and N-[1-(tert-butylaminocarbonyl)-1-phenylmethyl]-N-(4-methylphenyl)-3-(3-phenylsulfonyl-[1,2,5]oxadiazol-4-yl N-oxide)oxyphenyl carboxamide (6 d) exhibiting the strongest activity on SW1573 lung cell line (GI=110 and 21 nm) with selectivity indices of 70 and 470, respectively. Preliminary mechanistic studies suggest a relationship between NO release and antiproliferative activity. Our strategy allowed the rapid identification of at least two molecules as future candidates for the development of potent antitumor drugs.

[ES] La versatilidad de las aplicaciones de los hormigones reforzados con fibras (HRF) y los autocompactantes (HAC) en lugar del convencional los convierten en una alternativa de máximo interés para mejorar prestaciones y procesos industriales de elementos premoldeados existentes, mediante la aplicación de nuevos hormigones. Por ello se estudian ellos y la combinación de ambos (hormigón autocompactante con fibras, HACRF). Como referencia se tomó un H, usual en premoldeados locales, y en base a él se diseñó un HAC. Las variables analizadas fueron el tipo de fibras estructurales y su cuantía. Se estudiaron propiedades de los hormigones en estado fresco y endurecido. Los resultados obtenidos muestran claramente las diferencias de comportamiento en estado fresco y endurecido entre los hormigones estudiados. El empleo de HAC y la incorporación de fibras influyen significativamente en la reología, mejorando en estado endurecido la resistencia mecánica en relación al hormigón convencionalEn relación a la permeabilidad al aire y la resistencia a penetración de cloruros, los resultados obtenidos en todas las mezclas fueron similares.

El problema central de investigación de esta Tesis Doctoral es el estudio arqueológico de las lógicas que determinaron el uso y construcción del espacio por parte de grupos indígenas que habitaron las serranías de Uruguay. Concretamente, hemos focalizado en las construcciones de piedra monticulares y anulares encontradas en estos ambientes, denominadas en ámbitos locales como cairnes y vichaderos. Las evidencias sobre las que nos basamos fueron documentos y cartografía de época colonial e histórica y el registro arqueológico, analizados teniendo como base teórico-metodológica la Arqueología del Paisaje. El fenómeno de construir cairnes tiene una amplia dispersión a lo largo de las serranías del país, donde hemos reconocido cierta estandarización de morfologías, dimensiones, técnicas constructivas, configuraciones espaciales y emplazamientos, pero desconociéndose aún, al momento de iniciar esta investigación, su tipología, su funcionalidad, su cronología y su adscripción cultural. Para profundizar en estos aspectos hemos seleccionamos como caso de estudio a la sierra de Aguirre en el SE de Uruguay, un espacio acotado, abarcable para un estudio de estas características. Realizamos prospecciones arqueológicas en cumbres y planicies altas de la sierra y elegimos dos de las estructuras identificadas para realizar estudios de detalle y excavaciones arqueológicas. Los primeros trabajos realizados en sierra de Aguirre dejaron en evidencia un paisaje antropizado, que se manifiesta a través de la presencia de distintos tipos de arquitectura en piedra y transformación de espacios. La identificación del origen indígena de algunos tipos de estructuras en áreas serranas ha sido uno de los resultados principales. Estos grupos construyeron al menos dos tipos de estructuras de piedra y modificaron ciertos espacios de la sierra mediante transformaciones de bajo impacto.

Inspired by diverse marine bioactive compounds, the principle of molecular hybridization was applied to produce a series of new compounds combining diverse heterocyclic systems (oxa/thiazoles and tetrahydropyrans) via a triazole ring, attempting to increase the activity of individual building blocks. These new compounds exhibit a highly interesting antiproliferative activity against different human tumour cells and good selectivity when compared to normal cells. The formation of reactive oxygen species and the interaction with P-gp were also evaluated for the lead compounds.

Silver nanoparticles have particular properties that contribute to their very promising applications, novel in various fields of science, such as the development of biosensors, the diagnosis and treatment of cancer, the controlled release of drugs and the antimicrobial potential. The biological synthesis of nanoparticles is of great interest over other physical and chemical methods because the use of toxic chemicals and drastic reaction conditions are avoided. The extracellular biosynthesis using fungi could also make downstream processing much easier than the intracellular biosynthesis. One of the main applications of silver nanoparticles is their antimicrobial activity. Several studies have demonstrated the bactericidal properties of silver nanoparticles are different from silver ions, and that they are strongly influenced by their shape, size, concentration and colloidal state. In the present work, the ability of fungal strains from Uruguay to synthesize silver nanoparticles was studied. Eight fungi were able to synthesize nanoparticles. An extensive physicochemical characterization of the nanoparticles was carried out including ultraviolet-visible spectroscopy, transmission electron microscopy, dynamic light scattering, zeta-potential and gel electrophoretic mobility. According to the characterization and colloidal stability results, nanoparticles from three fungi were selected for antimicrobial activity assays. All nanoparticles were able to inhibit Escherichia coli growth, demonstrating their potential as effective antibacterial agent for use in biomedical applications.

Macrophages exert potent effector functions against invading microorganisms but constitute, paradoxically, a preferential niche for many bacterial strains to replicate. Using a model of infection by Salmonella Typhimurium, we have identified a molecular mechanism regulated by the nuclear receptor LXR that limits infection of host macrophages through transcriptional activation of the multifunctional enzyme CD38. LXR agonists reduced the intracellular levels of NAD+ in a CD38-dependent manner, counteracting pathogen-induced changes in macrophage morphology and the distribution of the F-actin cytoskeleton and reducing the capability of non-opsonized Salmonella to infect macrophages. Remarkably, pharmacological treatment with an LXR agonist ameliorated clinical signs associated with Salmonella infection in vivo, and these effects were dependent on CD38 expression in bone-marrow-derived cells. Altogether, this work reveals an unappreciated role for CD38 in bacterial-host cell interaction that can be pharmacologically exploited by activation of the LXR pathway.

A convenient, instrumentally simple, and efficient methodology to transform 1,2-dihydropyridines into benzoic esters is described. The generated multisubstituted benzoic esters feature different topologies spanning from simple aromatic rings to fused benzocycloalkane systems. As an extension of this methodology, these benzoic esters are efficiently transformed into an array of fluorenone frameworks featuring interesting and novel topological patterns.

Mitochondrial diseases are a group of clinically heterogeneous disorders that can be difficult to diagnose. We report a two and a half year old girl with clinical symptoms compatible with Leigh disease but with no definitive diagnosis. Using next generation sequencing we found that mutation 3697G > A was responsible for the patient's clinical symptoms. Corroboration was performed via segregation analysis in mother and sister and by evolutionary analysis that showed that the mutation is located in a highly conserved region across a wide range of species. Functional analyses corroborated the mutation effect and indicated that the pathophysiological alterations were partially restored by Coenzyme Q10. In addition, we proposed that the presence of the mutation at high frequencies causes the phenotype in the patient, while other family members with intermediate levels of heteroplasmy are symptoms-free.

Immobilization of antibodies (Ab) on hydrogels though long studied is still a challenge on account of the continuous development of new immune technologies. Enabling methodologies for antibody orientation, antibody stability and maximum recognition of their corresponding antigens is an object of intense study. Mini Ab such Fab fragments are less susceptible to conformational changes on surfaces, or unwanted reactivities compared to a whole Ab molecule. Herein, we have developed an immobilization protocol for a Fab anti epsilon toxin from Clostridium perfringens. Fine tuning of variables during immobilization showed a crucial role in the orientation and the spatial distribution of the Fab across the support surface. The final optimized immune-matrices demonstrated quantitative adsorption of antigen (1:1 molar ratio Fab to antigen) meaning that both the Fab biological activity was maintained after immobilization and an optimal orientation was achieved during the immobilization process. Immobilized Fab gained stability after immobilization as demonstrated by real time protein unfolding.

Biomimetic silica particles can be synthesized as a nanosized material within minutes in a process mimicked from living organisms such as diatoms and sponges. In this work, we have studied the effect of bovine serum albumin (BSA) as a template to direct the synthesis of silica nanoparticles (NPs) with the potential to associate proteins on its surface. Our approach enables the formation of spheres with different physicochemical properties. Particles using BSA as a protein template were smaller (∼250-380 nm) and were more monodisperse than those lacking the proteic core (∼700-1000 nm) as seen by dynamic light scattering (DLS), scanning electron microscopy (SEM), and environmental scanning electron microscopy (ESEM) analysis. The absence of BSA during synthesis produced silica nanoparticles without any porosity that was detectable by nitrogen adsorption, whereas particles containing BSA developed porosity in the range of 4 to 5 nm which collapsed on the removal of BSA, thus producing smaller pores. These results were in accordance with the pore size calculated by high-resolution transmission electron microscopy (HTEM). The reproducibility of the BSA-templated nanoparticle properties was determined by analyzing four batches of independent synthesizing experiments that maintained their properties. The high positive superficial charge of the nanoparticles facilitated adsorption under mild conditions of a range of proteins from an E. coli extract and a commercial preparation of laccase from Trametes versicolor. All of the proteins were quantitatively desorbed. Experiments conducted showed the reusability of the particles as supports for the ionic adsorption of the biomolecules. The protein loading capacity of the BSA-based biomimetic particles was determined using laccase as 98.7 ± 6.6 mg·g(-1) of particles.

Several breeding programs are under way to introduce resistance to bacterial wilt caused by Ralstonia solanacearum in solanaceous crops. The lack of screening methods allowing easy measurement of pathogen colonization and the inability to detect latent (i.e., symptomless) infections are major limitations when evaluating resistance to this disease in plant germplasm. We describe a new method to study the interaction between R. solanacearum and potato germplasm that overcomes these restrictions. The R. solanacearum UY031 was genetically modified to constitutively generate light from a synthetic luxCDABE operon stably inserted in its chromosome. Colonization of this reporter strain on different potato accessions was followed using life imaging. Bacterial detection in planta by this nondisruptive system correlated with the development of wilting symptoms. In addition, we demonstrated that quantitative detection of the recombinant strain using a luminometer can identify latent infections on symptomless potato plants. We have developed a novel, unsophisticated, and accurate method for high-throughput evaluation of pathogen colonization in plant populations. We applied this method to compare the behavior of potato accessions with contrasting resistance to R. solanacearum. This new system will be especially useful to detect latency in symptomless parental lines before their inclusion in long-term breeding programs for disease resistance.