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Nitric oxide (NO) is a gasotransmitter required in a broad range of mechanisms controlling plant development and stress conditions. However, little is known about the specific role of this signaling molecule during lipid storage in the seeds. Here, we show that NO is accumulated in developing embryos and regulates the fatty acid profile through the stabilization of the basic/leucine zipper transcription factor bZIP67. NO and nitro-linolenic acid target and accumulate bZIP67 to induce the downstream expression of FAD3 desaturase, which is misregulated in a non-nitrosylable version of the protein. Moreover, the post-translational modification of bZIP67 is reversible by the trans-denitrosylation activity of peroxiredoxin IIE and defines a feedback mechanism for bZIP67 redox regulation. These findings provide a molecular framework to control the seed fatty acid profile caused by NO, and evidence of the in vivo functionality of nitro-fatty acids during plant developmental signaling.

Prostate cancer is the second most frequent cancer in men worldwide after lung cancer. Its diagnosis is based on the identification of the Gleason score that evaluates the abnormality of cells in glands through the analysis of the different Gleason patterns within tissue samples. The recent advancements in computational pathology, a domain aiming at developing algorithms to automatically analyze digitized histopathology images, lead to a large variety and availability of datasets and algorithms for Gleason grading and scoring. However, there is no clear consensus on which methods are best suited for each problem in relation to the characteristics of data and labels. This paper provides a systematic comparison on nine datasets with state-of-the-art training approaches for deep neural networks (including fully-supervised learning, weakly-supervised learning, semi-supervised learning, Additive-MIL, Attention-Based MIL, Dual-Stream MIL, TransMIL and CLAM) applied to Gleason grading and scoring tasks. The nine datasets are collected from pathology institutes and openly accessible repositories. The results show that the best methods for Gleason grading and Gleason scoring tasks are fully supervised learning and CLAM, respectively, guiding researchers to the best practice to adopt depending on the task to solve and the labels that are available.

Background: Among all the cancers known today, prostate cancer is one of the most commonly diagnosed in men. With modern advances in medicine, its mortality has been considerably reduced. However, it is still a leading type of cancer in terms of deaths.The diagnosis of prostate cancer is mainly conducted by biopsy test. From this test, Whole Slide Images are obtained, from which pathologists diagnose the cancer according to the Gleason scale. Within this scale from 1 to 5, grade 3and above is considered malignant tissue. Several studies have shown an inter- observer discrepancy between pathologists in assigning the value of the Gleason scale. Due to the recent advances in artificial intelligence, its application to the computational pathology field with the aim of supporting and providing a second opinion to the professional is of great interest. Method:In this work, the inter-observer variability of a local dataset of 80 whole-slide images annotated by a team of 5 pathologists from the same group was analyzed at both area and label level. Four approaches were followed to train six different Convolutional Neural Network architectures, which were evaluated on the same data set on which the inter-observer variability was analyzed. Results: An inter-observer variability of 0.6946k was obtained, with 46% discrepancy in terms of area size of the annotations performed by the pathologists. The best trained models achieved 0.826±0.014k on the test set when trained with data from the same source. Conclusions: The obtained results show that deep learning-based automatic diagnosi s systems could help reduce the widely-known inter-observer variability that is present among pathologists and support them in their decision, serving as a second opinion or as a triage tool for medical centers.

The plantation of tree islets within deforested areas (applied nucleation) has been proposed as a restoration strategy able to catalyze tree colonization in the matrix with reasonable costs and labor compared to traditional restoration methods such as extensive plantations. However, recent work has questioned its effectiveness in comparison to natural forest regeneration, and observational studies have eventually reported a limited expansion of tree islets after years of planting. To infer the processes affecting the effectiveness of applied nucleation, we used a stochastic model able to predict avian movement and fruit consumption, tree seed dispersal and seedling establishment in a fragmented temperate forest landscape of northern Spain. First, we simulated scenarios of natural regeneration and compared them to applied nucleation in terms of tree colonization in the remaining deforested matrix. Subsequently, in applied nucleation scenarios we assessed the effects of islet size and matrix area recovered on tree recruitment into the matrix. Also, we evaluated if such effects were mediated by the scale of movement of frugivores. Our results showed that applied nucleation triggers matrix colonization by trees at higher rates than natural regeneration because it avoids frugivore entrapment within forest edges. Also, that large tree islets can hamper matrix colonization despite being visited more frequently. If large tree islets offer abundant resources, animals do not need to leave their boundaries, concentrating seed deposition within islets. As expected, such effects depended on the scale of movement of frugivores. Our work highlights that in nucleated landscapes, islets harboring fruit-producing trees should attract animal dispersers, and also foster animal inter-islet movement. Both conditions should be met to ensure seed deposition and seedling establishment in the deforested matrix, promoting forest recovery independent of islet expansion through vegetative growt

In this paper, we investigate Astronomical Observations Scheduling which is a type of Multi-Objective Combinatorial Optimization Problem, and detail its specific challenges and requirements and propose the Hybrid Accumulative Planner (HAP), a hybrid multi-start metaheuristic scheduler able to adapt to the different variations and demands of the problem. To illustrate the capabilities of the proposal in a real-world scenario, HAP is tested on the Atmospheric Remote-sensing Infrared Exoplanet Large-survey (Ariel) mission of the European Space Agency (ESA), and compared with other studies on this subject including an Evolutionary Algorithm (EA) approach. The results show that the proposal outperforms the other methods in the evaluation and achieves better scientific goals than its peers. The consistency of HAP in obtaining better results on the available datasets for Ariel, with various sizes and constraints, demonstrates its competence in scalability and adaptability to different conditions of the problem.

In recent years, locomotion mechanisms exhibited by vertebrate animals have been the inspiration for the improvement in the performance of robotic systems. These mechanisms include the adaptability of their locomotion to any change registered in the environment through their biological sensors. In this regard, we aim to replicate such kind of adaptability through a sCPG. This sCPG generates different loco motion (rhythmic) patterns which are driven by an external stimulus, that is, the output of a FSR sensor to provide feedback. The sCPG consists of a network of five populations of LIF neurons designed with a specific topology in such a way that the rhythmic patterns can be generated and driven by the aforemen tioned external stimulus. Therefore, eventually, the locomotion of an end robotic platform could be adapted to the terrain by using any sensor as input. The sCPG with adaptation has been numerically val idated at software and hardware level, using the Brian 2 simulator and the SpiNNaker neuromorphic plat form for the latest. In particular, our experiments clearly show an adaptation in the oscillation frequencies between the spikes produced in the populations of the sCPG while the input stimulus varies. To validate the robustness and adaptability of the sCPG, we have performed several tests by variating the output of the sensor. These experiments were carried out in Brian 2 and SpiNNaker; both implementa tions showed a similar behavior with a Pearson correlation coefficient of 0.905

The correct identi cation of burst events is crucial in many scenarios, ranging from basic neuroscience to biomedical applications. However, none of the burst detection methods that can be found in the literature have been widely adopted for this task. As an alternative to conventional techniques, a novel neuromorphic approach for real-time burst detection is proposed and tested on acquisitions from in vitro cultures. The system consists of a Neuromorphic Auditory Sensor, which converts the input signal obtained from electrophysiological recordings into spikes and decomposes them into di erent frequency bands. The output of the sensor is sent to a trained spiking neural network implemented on a SpiNNaker board that discerns between bursting and non-bursting activity. This data-driven approach was compared with 8 di erent conventional spike-based methods, addressing some of their drawbacks, such as being able to detect both high and low frequency events and working in an online manner. Similar results in terms of number of detected events, mean burst duration and correlation as current state-ofthe- art approaches were obtained with the proposed system, also bene ting from its lower power consumption and computational latency. Therefore, our neuromorphic-based burst detection paves the road to future implementations for neuroprosthetic applications.

Prostate cancer (PCa) is one of the most commonly diagnosed cancer and one of the leading causes of death among men, with almost 1.41 million new cases and around 375,000 deaths in 2020. Artificial Intelligence algorithms have had a huge impact on medical image analysis, including digital histopathology, where Convolutional Neural Networks (CNNs) are used to provide a fast and accurate diagnosis, supporting experts in this task. To perform an automatic diagnosis, prostate tissue samples are first digitized into gigapixel-resolution whole-slide images. Due to the size of these images, neural networks cannot use them as input and, therefore, small subimages called patches are extracted and predicted, obtaining a patch-level classification. In this work, a novel patch aggregation method based on a custom Wide & Deep neural network model is presented, which performs a slide-level classification using the patch-level classes obtained from a CNN. The malignant tissue ratio, a 10-bin malignant probability histogram, the least squares regression line of the histogram, and the number of malignant connected components are used by the proposed model to perform the classification. An accuracy of 94.24% and a sensitivity of 98.87% were achieved, proving that the proposed system could aid pathologists by speeding up the screening process and, thus, contribute to the fight against PCa.

OpenNAS is an open-source tool for automatically generating the source files to create a Neuromorphic Auditory Sensor (NAS) VHDL project for FPGA. OpenNAS guides the user with a friendly interface that allows configuring the NAS’ parameters using a five-step wizard for code generation. OpenNAS provides support to several audio input interfaces (AC’97 audio codec, I2S-ADC and PDM microphones), different processing architectures (cascade and parallel), and neuromorphic output interfaces (parallel AER, SpiNNaker). After NAS generation, users have everything ready for building, simulating, and synthesizing the VHDL project for a target FPGA. OpenNAS is fully modular, which allows providing support to new features in an easy way

Visualizing the output from event-based sensors and analyzing the information within it is crucial when developing new neuromorphic systems. A novel open-source, cross-platform Python package called pyNAVIS is proposed, which provides a set of functionalities to analyze and process spiking information obtained from neuromorphic cochleas, along with other tools to generate files and datasets for machine learning tasks. This modular package can be integrated in complex and higher-level projects, which could be useful for researchers working in neuromorphic audio processing tasks.

Objective: To reconstruct the etiology of a perimortem injury observed on a Neolithic – Chalcolithic cranium (5060 – 4400 yrs cal. BP). Materials: A cranium of an old adult male individual belonging to a collective burial from Cova Foradada site (Calafell, Tarragona, Spain). Methods: The cranium was analyzed macroscopically and microscopically using micro-CT scan. Results: The fracture on the right parietal bone presents characteristics of perimortem trauma. The morphology of the point of impact allows for the interpretation of this fracture as a result of impact by an object with a straight and sharp edge. In addition, the same cranium presents two healed antemortem injuries. Conclusions: The object that most likely caused the cranial fracture was a stone adze. The blow occurred from behind the individual, possibly by a right-handed attacker. Significance: The potential to link cranial fractures with specific tools increases our understanding of interpersonal violence during the Neolithic. Limitations: It is not possible to infer if this cranial injury was related to a large-scale intergroup confrontation or an intragroup violent event. Suggestions for further research: To investigate additional similar sites in the northeastern Iberian Peninsula to better characterize the fracture pattern caused by stone axes and adzes as well as other objects used as weapons during the Neolithic – Chalcolithic ages.

This study presents a new cyanobacterium of the genus Geitlerinema sp. isolated from the Chilean coastal desert as a rich source of bioactive compounds. Phylogenetic tree is shown along with a broad biochemical characterization using conventional and pressurized liquid extraction. Variables as extraction yield, phycobiliproteins, lipids and methyl palmitate (C16:0) were studied by means of two experimental designs based on a response surface methodology (RSM) employing two factorial design 32. Factors as temperature (20–200 °C) and percentage of solvents were selected resulting in Factorial Design A with 0–100% (v/v) of ethanol–water and Factorial Design B with 0–100 (v/v) of ethanol–limonene solvents. The main bioactive compounds were total proteins (∼52% wt), phycobiliproteins (especially C-phycocyanin) and methyl palmitate (C16:0). Data obtained showed that water and ethanol:water (1:1, v/v) improved the extraction yield and phycobiliproteins recovery. As for ethanol:limonene extracts, they were better for lipids and methyl palmitate extractions. In general, high temperatures only improved the extraction yield whereas low temperatures favor the extraction of phycobiliproteins. Therefore, results indicated that Geitlerinema sp. could be one of the several cyanobacteria genera that display functional properties for nutritional, pharmaceutical or industrial use as sustainable and cost-effective bioactive compounds in algal biorefinery model.

Lacustrine coasts, especially those associated with shallow highly fluctuating lakes, do not evolve in exactly the same way as marine coasts. Climate variability can significantly modify the morphology of the water body, affecting the lake surface area, the available fetch, and the water depth, resulting in possible changes to wave generation and propagation. To look more closely at the nature of coastal sediments generated under such specific environments, we selected examples of both shallow lacustrine and open ocean beach deposits. The first location was Gallocanta Lake, a shallow body of water in the Iberian Chain in central Spain; and the second was Mazagón Beach, in the Gulf of Cádiz, a mesotidal sandy beach exposed to Atlantic waves and tides. Both deposits are formed by beach and ridge and runnel systems. The sedimentary structures of the two deposits were compared, together with a genetic scheme for these. In both cases, applying wave propagation models facilitated the interpretation of the sedimentary processes responsible for their genesis. Although the general morphosedimentary characteristics of the two deposits are roughly similar, a more detailed analysis revealed significant differences, some of which could be used as diagnostic indicators of shallow lacustrine environments in geoarchives. Effective fetch seemed not to be a determining factor for explaining the textural differences between the two deposits. Local factors such as wind strength and direction, and approaching wave refraction patterns are important in controlling the final energy reaching both types of coasts, and, hence, the resulting geometry of the clastic coastal deposits. The type and maturity of the available sediment also influence the final textural properties of the deposits. Open ocean beaches receive sporadic high energetic storms; these partially erode the sedimentary record and reduce the preservation potential of coastal deposits compared with those from shallow lake beache

Initially, robots were developed with the aim of making our life easier, carrying out repetitive or dangerous tasks for humans. Although they were able to perform these tasks, the latest generation of robots are being designed to take a step further, by performing more complex tasks that have been carried out by smart animals or humans up to date. To this end, inspiration needs to be taken from biological examples. For instance, insects are able to optimally solve complex environment navigation problems, and many researchers have started to mimic how these insects behave. Recent interest in neuromorphic engineering has motivated us to present a real-time, neuromorphic, spike-based Central Pattern Generator of application in neurorobotics, using an arthropod-like robot. A Spiking Neural Network was designed and implemented on SpiNNaker. The network models a complex, online-change capable Central Pattern Generator which generates three gaits for a hexapod robot locomotion. Recon gurable hardware was used to manage both the motors of the robot and the real-time communication interface with the Spiking Neural Networks. Real-time measurements con rm the simulation results, and locomotion tests show that NeuroPod can perform the gaits without any balance loss or added delay.

Early embryonic development is driven exclusively by maternal gene products deposited into the oocyte. Although critical in establishing early developmental programs, maternal gene functions have remained elusive due to a paucity of techniques for their systematic disruption and assessment. CRISPR-Cas13 systems have recently been employed to degrade RNA in yeast, plants, and mammalian cell lines. However, no systematic study of the potential of Cas13 has been carried out in an animal system. Here, we show that CRISPR-RfxCas13d (CasRx) is an effective and precise system to deplete specific mRNA transcripts in zebrafish embryos. We demonstrate that zygotically expressed and maternally provided transcripts are efficiently targeted, resulting in a 76% average decrease in transcript levels and recapitulation of well-known embryonic phenotypes. Moreover, we show that this system can be used in medaka, killifish, and mouse embryos. Altogether, our results demonstrate that CRISPR-RfxCas13d is an efficient knockdown platform to interrogate gene function in animal embryos.

Here we present a novel G4-binding family of compounds based on a central core of phenyl ditriazole (PDTZ) modified with carbohydrates and phenyl pyrrolidinyl side-chains. Their synthesis was achieved using controlled click chemistry conditions to obtain both, symmetric and dissymmetric carb-PDTZ derivatives without any intermediate protecting steps through an optimized methodology. Binding of the new carb-PDTZ to a variety of G-quadruplex motifs was examined using different biophysical techniques. The symmetric carb-PDTZ derivatives were not able to stabilize G4, but the dissymmetric ones (containing one sugar and one phenyl pyrrolidinyl side-chain) did. Interestingly, the dissymmetric carb-PDTZ derivatives showed much higher G4 vs duplex DNA selectivity than the control compound PDTZ 1, which contains two phenyl pyrrodilinyl side-chains and no carbohydrates. Their potential antitumoral activity was also investigated by in vitro cytotoxicity measurements on different cancerous cell lines. All carb-PDTZ derivatives showed higher IC values than the control PDTZ 1, probably due to the lack of compound stability of some derivatives and to lower cellular uptake.

Introduction: The simple phenol hydroxytyrosol (OHTyr) has been associated with the beneficial health effects of extra virgin olive oil. Pre-clinical studies have identified Tyr hydroxylation, mediated by cytochrome P450 isoforms CYP2A6 and CYP2D6, as an additional source of OHTyr. Aim: We aimed to (i) confirm Tyr to OHTyr bioconversion in vivo in humans, (ii) assess the cardiovascular benefits of this bioconversion, and (iii) determine their interaction with a polygenic activity score (PAS) from CYP2A6 and CYP2D6 genotypes. Methods: Randomized, crossover, controlled study. Individuals at cardiovascular risk (n = 33) received: white wine (WW) (females 1, males 2 standard drinks/day), WW plus Tyr capsules (WW + Tyr) (25 mg Tyr capsule, one per WW drink), and water (control) ad libitum. Participants were classified by a PAS as low versus normal activity metabolizers. Results: OHTyr recovery following WW + Tyr was higher than after other interventions (P < 0.05). Low PAS individuals had lower OHTyr/Tyr ratios compared to individuals with normal PAS. WW + Tyr improved endothelial function, increased plasma HDL-cholesterol and antithrombin IIII, and decreased plasma homocysteine, endothelin 1, and CD40L, P65/RELA, and CFH gene expression in peripheral blood mononuclear cells (p < 0.05). Combining Tyr capsule(s) with WW abolished the increase in iNOS, eNOS, VEGFA, and CHF expressions promoted by WW (p < 0.05). Conclusions: Tyr, and its partial biotransformation into OHTyr, promoted cardiovascular health-related benefits in humans after dietary doses of Tyr. The study design allowed the health effects of individual phenols to be singled out from the dietary matrix in which they are naturally found.

Here we present new and original data on the endogenous conversion of tyrosol (Tyr) into hydroxytyrosol (OHTyr) in humans and its effects on the cardiovascular system. A randomized, crossover, controlled clinical trial was performed with individuals at cardiovascular risk (n = 33). They received white wine (WW) (females 1, males 2 standard drinks/day), WW plus Tyr capsules (WW + Tyr) (25mg Tyr capsule, one per WW drink), and water (control) ad libitum. Intervention periods were of 4 weeks preceded by three-week wash-out periods. We assessed the conversion of Tyr to OHTyr, its interaction with a polygenic activity score (PAS) from CYP2A6 and CYP2D6 genotypes, and the effects on cardiovascular risk markers. For further details and experimental findings please refer to the article >Cardiovascular benefits of tyrosol and its endogenous conversion into hydroxytyrosol in humans. A randomized, controlled trial> [1].

The neglected tropical diseases Human African Trypanosomiasis and leishmaniasis are caused by infection with trypanosomatid parasites Trypanosoma brucei and Leishmania spp, respectively. The genomes of these organisms contain multiple putative G-quadruplex (G4) forming sequences which have recently been proposed to mediate processes relevant for parasite survival. Therefore, G4 could be considered as potential targets for a novel approach towards the development of antiparasitic drugs. Recently, we have demonstrated that G4 ligands such as carbohydrate naphthalene diimide conjugates (carb-NDIs) possess notable antiparasitic activity. Herein, we have synthesized a new family of carb-NDIs, characterized by significant structural variability, and evaluated their anti-parasitic activity, with special focus on T brucei. The interaction with relevant G4 sequences was evaluated in vitro through independent biophysical methods (FRET melting assays under competing conditions with double stranded DNA, circular dichroism and fluorescence titrations). Finally, flow cytometry and confocal microscopy experiments demonstrated that the conjugates exhibit excellent uptake into T. brucei parasites, localizing in the nuclei and kinetoplasts. Promising antiparasitic activity and selectivity against control mammalian cells, together with their peculiar mechanism of action, render the carb-NDI conjugates as suitable candidates for the development of an innovative treatment of trypanosomiasis. (C) 2018 Elsevier Masson SAS. All rights reserved.

© 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0, Here we present new and original data on the endogenous con-version of tyrosol (Tyr) into hydroxytyrosol (OHTyr) in humans and its effects on the cardiovascular system. A randomized,crossover, controlled clinical trial was performed with individuals at cardiovascular risk (n¼33). They received white wine (WW)(females 1, males 2 standard drinks/day), WW plus Tyr capsules(WWþTyr) (25mg Tyr capsule, one per WW drink), and water(control)ad libitum. Intervention periods were of 4 weeks pre-ceded by three-week wash-out periods. We assessed the conversion of Tyr to OHTyr, its interaction with a polygenic activity score(PAS) fromCYP2A6andCYP2D6genotypes, and the effects on cardiovascular risk markers. For further details and experimental findings please refer to the article“Cardiovascular benefits of tyrosol and its endogenous conversion into hydroxytyrosol Inhumans. A randomized, controlled trial”, Peer Reviewed, Postprint (author's final draft)

Recent biological studies have focused on understanding animal interactions and welfare. To help biolo- gists to obtain animals’ behavior information, resources like wireless sensor networks are needed. More- over, large amounts of obtained data have to be processed off-line in order to classify different behaviors.There are recent research projects focused on designing monitoring systems capable of measuring someanimals’ parameters in order to recognize and monitor their gaits or behaviors. However, network unre- liability and high power consumption have limited their applicability.In this work, we present an animal behavior recognition, classification and monitoring system based ona wireless sensor network and a smart collar device, provided with inertial sensors and an embeddedmulti-layer perceptron-based feed-forward neural network, to classify the different gaits or behaviorsbased on the collected information. In similar works, classification mechanisms are implemented in aserver (or base station). The main novelty of this work is the full implementation of a reconfigurableneural network embedded into the animal’s collar, which allows a real-time behavior classification andenables its local storage in SD memory. Moreover, this approach reduces the amount of data transmittedto the base station (and its periodicity), achieving a significantly improving battery life. The system hasbeen simulated and tested in a real scenario for three different horse gaits, using different heuristics andsensors to improve the accuracy of behavior recognition, achieving a maximum of 81%.

This software presents diverse utilities to perform the first post-processing layer taking the neuromorphic auditory sensors (NAS) information. The used NAS implements in FPGA a cascade filters architecture, imitating the behavior of the basilar membrane and inner hair cells and working with the sound information decomposed into its frequency components as spike streams. The well-known neuromorphic hardware interface Address-Event-Representation (AER) is used to propagate auditory information out of the NAS, emulating the auditory vestibular nerve. Using the information packetized into aedat files, which are generated through the jAER software plus an AER to USB computer interface, NAVIS implements a set of graphs that allows to represent the auditory information as cochleograms, histograms, sonograms, etc. It can also split the auditory information into different sets depending on the activity level of the spike streams. The main contribution of this software tool is that it allows complex audio post-processing treatments and representations, which is a novelty for spike-based systems in the neuromorphic community and it will help neuromorphic engineers to build sets for training spiking neural networks (SNN).

Recent studies in animal systems have shown that NO can interact with fatty acids to generate nitro-fatty acids (NO-FAs). They are the product of the reaction between reactive nitrogen species and unsaturated fatty acids, and are considered novel mediators of cell signaling based mainly on a proven anti-inflammatory response. Although these signaling mediators have been described widely in animal systems, NO-FAs have scarcely been studied in plants. Preliminary data have revealed the endogenous presence of free and protein-adducted NO-FAs in extra-virgin olive oil (EVOO), which appear to be contributing to the cardiovascular benefits associated with the Mediterranean diet. Importantly, new findings have displayed the endogenous occurrence of nitro-linolenic acid (NO-Ln) in the model plant Arabidopsis thaliana and the modulation of NO-Ln levels throughout this plant's development. Furthermore, a transcriptomic analysis by RNA-seq technology established a clear signaling role for this molecule, demonstrating that NO-Ln was involved in plant-defense response against different abiotic-stress conditions, mainly by inducing the chaperone network and supporting a conserved mechanism of action in both animal and plant defense processes. Thus, NO-Ln levels significantly rose under several abiotic-stress conditions, highlighting the strong signaling role of these molecules in the plant-protection mechanism. Finally, the potential of NO-Ln as a NO donor has recently been described both in vitro and in vivo. Jointly, this ability gives NO-Ln the potential to act as a signaling molecule by the direct release of NO, due to its capacity to induce different changes mediated by NO or NO-related molecules such as nitration and S-nitrosylation, or by the electrophilic capacity of these molecules through a nitroalkylation mechanism. Here, we describe the current state of the art regarding the advances performed in the field of NO-FAs in plants and their implication in plant physiology.

Analysis of the geological record has made it possible to delimit for the Guadalquivir estuary the traces of extreme wave events (EWEs) during the Roman period in the Iberian Peninsula (218 BCE to 476 CE). The largest event occurred in the 2nd-3rd century CE. It generated clearly visible erosive effects in the coastal barriers, including washover fans and erosional scarps. In the inner estuary, however, the effects were minor: crevasse splays that broke levees and cheniers, as well as a residual sedimentary lag. The significant development of the spits protected the inner estuary from the marine incursion, which only caused a water level rise with low-regime waves. Correlation of the geomorphological and sedimentary marks left by this event with the archaeological and geological evidence of other events recognized elsewhere in the Gulf of Cadiz effectively argues for a tsunami as to the nature of the 2nd-3rd century CE event. Yet this and the other identified EWEs in the Guadalquivir estuary during the pre-Roman and the Roman period all fit a model of paleogeographic evolution dominated by processes of coastal progradation and estuarine infilling. Radiocarbon dating, geomorphological analysis, and historical references fail to warrant the so-called '218-209 BCE' Atlantic tsunami, as hypothesized in the received scientific literature. In pre-Roman and Roman times, human occupation at the mouth of the Guadalquivir River was strongly influenced by various geodynamic processes, the location of the settlements being contingent upon dependable, fast communication with the sea and, above all, upon adequate protection from EWEs, on the leeward side of spits. Progressive progradation of these coastal barriers combined with the gradual infilling of the estuary to make navigation to open sea increasingly difficult and, eventually, to result in the abandonment of settlements.

Demographic and genetic connectivity of fragmented plant populations will depend on effective propagule flow across the landscape. We analyze functional connectivity in a holm oak (Quercus ilex) fragmented landscape by considering three important stages driving recruitment: effective pollination, acorn production and acorn dispersal. We used a network approach to (1) determine if pollen-mediated gene exchange across the landscape was spatially structured; (2) estimate the effects of limited acorn dispersal on functional connectivity; (3) identify which landscape traits could drive source–sink dynamics of gene flow. Although long distance dispersal was relatively frequent, most effective pollen flow occurred over short distances (<100 m). This resulted in a significantly modular structure of the mating network, yielding higher gene flow among nearby fragments. Limited mouse acorn hoarding activity had a strong impact on landscape connectivity, decreasing male gametic immigration rates into forest patches by one order of magnitude Besides, our results show that big forest fragments (>10 ha) are the main pollen sources, while small ones (<1 ha) are important pollen sinks. Thus, big fragments are critical to maintain functional connectivity, while small forest fragments may provide acorn crops better representing regional genetic diversity. In addition to area effects, less isolated and more central fragments showed higher migration rates and exchanged effective pollen with more fragments. Hence, we expected that landscapes with uniform or clumped distribution of big forest fragments would show optimal connectivity traits. However, despite that simulated gene flow was more evenly distributed across the landscape, connectance and migration rates decreased. Our results call for caution before translating patch-level management guidelines to the landscape scale. They also show that the level of functional connectivity may change throughout the recruitment process, suggesti

Background: Guanine-rich oligonucleotides are capable of forming tetrahelical structures known as G-quadruplexes with interesting biological properties. We have investigated the effects of site-specific substitution in the loops and in the tetrads model G-quadruplexes using thymine glycol nucleic acid (GNA) units, . l-thymidine and 8-Br-2'-deoxyguanosine. Methods: Modified oligonucleotides were chemically synthesized and spectroscopic techniques were used to determine the relative stability of the modified G-quadruplex. The double 8-BrdG-modified quadruplexes were further characterized by Nuclear Magnetic Resonance. Binding to thrombin of selected quadruplex was analyzed by gel electrophoresis retention assay. Results: The most interesting results were found with a 8-bromoG substitution that had the larger stabilization of the quadruplex. NMR studies indicate a tight relationship between the loops and the tetrads to accommodate 8-bromoG modifications within the TBA. Conclusions: The substitutions of loop positions with GNA T affect the TBA stability except for single modification in T7 position. Single . l-thymidine substitutions produced destabilization of TBA. Larger changes on quadruplex stability are observed with the use of 8-bromoG finding a single substitution with the highest thermal stabilization found in thrombin binding aptamers modified at the guanine residues and having good affinity for thrombin. Double 8-BrdG modification in . anti positions of different tetrads produce a conformational flip from . syn to . anti conformation of 8-Br-dG to favor loop-tetrad interaction and preserve the overall TBA stability. General significance: Modified guanine-rich oligonucleotides are valuable tools for the search for G-quadruplex structures with higher thermal stability and may provide compounds with interesting protein-nucleic acid binding properties. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Danie

The antioxidant activity of gallic acid and a series of alkyl gallates (C4-C18) and glycosylated alkyl gallates (C4-C18) on fish oil-in-water emulsions was studied. Three types of emulsifiers, lecithin, Tween-20 and sodium dodecyl sulphate (SDS) were tested. A nonlinear behavior of the antioxidant activity of alkyl gallates when increasing alkyl chain length was observed for emulsions prepared with lecithin. Medium-size alkyl gallates (C6-C12) were the best antioxidants. In contrast, for emulsions prepared with Tween-20, the antioxidants seem to follow the polar paradox. Glucosyl alkyl gallates were shown previously to be better surfactants than alkyl gallates. Nevertheless, they exhibited a worse antioxidant capacity than their corresponding alkyl gallates, in emulsions prepared with lecithin or Tween-20, indicating the greater relevance of having three OH groups at the polar head in comparison with having improved surfactant properties but just a di-ortho phenolic structure in the antioxidant.

Oligoribonucleotide conjugates carrying apolar carbohydrates at the 5′-end and the corresponding siRNA duplexes have been prepared using phosphoramidite chemistry. All the carbohydrate-siRNA derivatives were compatible with RNA interference machinery if transfected with oligofectamine. In the absence of a transfection agent, some of them exerted certain reduction of gene expression. Double-tailed permethylated glucose conjugated to siRNA through a long spacer inhibited gene expression up to 26% compared to the scrambled duplex. Such modifications contribute positively to the stability of oligoribonucleotides against 5′-exonuclease degradation.

Oligoribonucleotide conjugates carrying apolar carbohydrates at the 5′-end and the corresponding siRNA duplexes have been prepared using phosphoramidite chemistry. All the carbohydrate-siRNA derivatives were compatible with RNA interference machinery if transfected with oligofectamine. In the absence of a transfection agent, some of them exerted certain reduction of gene expression. Double-tailed permethylated glucose conjugated to siRNA through a long spacer inhibited gene expression up to 26% compared to the scrambled duplex. Such modifications contribute positively to the stability of oligoribonucleotides against 5′-exonuclease degradation.

Polarized trafficking of adhesion receptors plays a pivotal role in controlling cellular behavior during morphogenesis. Particularly, clathrin-dependent endocytosis of integrins has long been acknowledged as essential for cell migration. However, little is known about the contribution of integrin trafficking to epithelial tissue morphogenesis. Here we show how the transmembrane protein Opo, previously described for its essential role during optic cup folding, plays a fundamental role in this process. Through interaction with the PTB domain of the clathrin adaptors Numb and Numbl via an integrin-like NPxF motif, Opo antagonizes Numb/Numbl function and acts as a negative regulator of integrin endocytosis in vivo. Accordingly, numb/numbl gain-of-function experiments in teleost embryos mimic the retinal malformations observed in opo mutants. We propose that developmental regulator Opo enables polarized integrin localization by modulating Numb/Numbl, thus directing the basal constriction that shapes the vertebrate retina epithelium.

Isoflavones are a group of substances that belong to the family of phytoestrogens. These natural substances may offer several benefits to human health. One of the most important sources for human isoflavone intake is soy and soybean food derivatives. An improved sample treatment followed by a high performance liquid chromatographic method for the determination of isoflavones in supplemented milk and juices is proposed and compared to the AOAC official method. Detection limits found were between 0.2 and 0.3 mg L−1 for daidzein and genistein respectively. Quantification limits found were between 0.7 mg L−1 for daidzein to 1.0 mg L−1 for genistin, while inter and intra-day variability was under 10% in all cases. Recoveries for spiked samples were over 90% and under 110%. The method was validated by comparison with the AOAC method and by recovery assay methodology.

Ojoplano (Opo) is a morphogenetic gene playing an important role during embryogenesis in medaka. This report focuses on the identification and characterization of the mouse Opo gene. We examined Opo expression by whole-mount in situ hybridization and in situ hybridization on sagittal sections during mouse embryogenesis. First expression in whole-mounts was detected at Theiler stages 15–17 (E 9.5–10.5 dpc) as a spotted specific staining in migrating neural crest cells and in placodal structures. A complex expression pattern was observed in Theiler stage 22–23 (E 14.5 dpc) in sagittal sections, including expression in skeletal structures (skull, vertebrae, ribs, bones of the locomotor system), in the nasal region, the heart and the eye. Fusion proteins revealed the localization of OPO within the cytoplasm with a reticular distribution that largely overlapped with the endoplasmic reticulum. Opo shows homology to human transcripts linked to a hereditary craniofacial malformation, orofacial cleft 1 (OFC1). The expression of mouse Opo in neural crest derivatives and skull elements further supports this link.

For over a century, the vertebrate eye has served as a paradigm for organogenesis. It forms through a complex sequence of morphogenetic events, involving the lateral evagination of the optic vesicles and their subsequent folding into the optic cups. Through intensive studies by experimental embryologists, anatomical descriptions of the process were available since many decades. Recent genetic and molecular work has illuminated essential features of the stereotyped cellular behaviour driving eye morphogenesis. The first pieces of the molecular machinery operating in each individual progenitor cell have been identified. These studies now set the groundwork for a system-wide approach towards understanding the cellular and molecular mechanisms involved in shaping the vertebrate eye.