Computational Principles of Supervised Learning in the Cerebellum.

Supervised learning plays a key role in the operation of many biological and artificial neural networks. Analysis of the computations underlying supervised learning is facilitated by the relatively simple and uniform architecture of the cerebellum, a brain area that supports numerous motor, sensory, and cognitive functions. We highlight recent discoveries indicating that the cerebellum implements supervised learning using the following organizational principles: ( a) extensive preprocessing of input representations (i.e., feature engineering), ( b) massively recurrent circuit architecture, ( c) linear input-output computations, ( d) sophisticated instructive signals that can be regulated and are predictive, ( e) adaptive mechanisms of plasticity with multiple timescales, and ( f) task-specific hardware specializations. The principles emerging from studies of the cerebellum have striking parallels with those in other brain areas and in artificial neural networks, as well as some notable differences, which can inform future research on supervised learning and inspire next-generation machine-based algorithms.

Dendritic Inhibition by Shh Signaling-Dependent Stellate Cell Pool Is Critical for Motor Learning.

Cerebellar inhibitory interneurons are important regulators of neural circuit activity for diverse motor and nonmotor functions. The molecular layer interneurons (MLIs), consisting of basket cells (BCs) and stellate cells (SCs), provide dendritic and somatic inhibitory synapses onto Purkinje cells, respectively. They are sequentially generated in an inside-out pattern from Pax2+ immature interneurons, which migrate from the prospective white matter to the ML of the cortex. However, little is known about how MLI subtype identities and pool sizes are determined, nor are their contributions to motor learning well understood. Here, we show that GABAergic progenitors fated to generate both BCs and SCs respond to the Sonic hedgehog (Shh) signal. Conditional abrogation of Shh signaling of either sex inhibited proliferation of GABAergic progenitors and reduced the number of Pax2+ cells, whereas persistent Shh pathway activation increased their numbers. These changes, however, did not affect early born BC numbers but selectively altered the SC pool size. Moreover, genetic depletion of GABAergic progenitors when BCs are actively generated also resulted in a specific reduction of SCs, suggesting that the specification of MLI subtypes is independent of Shh signaling and their birth order and likely occurs after Pax2+ cells settle into their laminar positions in an inside-out sequence. Mutant mice with reduced SC numbers displayed decreased dendritic inhibitory synapses and neurotransmission onto Purkinje cells, resulting in an impaired acquisition of eyeblink conditioning. These findings also reveal an essential role of Shh signaling-dependent SCs in regulating inhibitory dendritic synapses and motor learning.SIGNIFICANCE STATEMENT The cerebellar circuit that enables fine motor learning involves MLIs of BCs and SCs, which provide dendritic and somatic inhibitory synapses onto Purkinje cells. Little is known about how their identities and numbers are determined, nor are their specific contributions to motor learning well understood. We show that MLI subtypes are specified independent of Shh signaling and their birth orders but appear to occur in their terminal laminar positions according to the inside-out sequence. This finding challenges the current view that MLI subtypes are specified sequentially at the progenitor level. We also demonstrate that dendritic inhibition by Shh signaling-dependent SC pool is necessary for motor learning.

Relevance of well-being, resilience, and health-related quality of life to mental health profiles of European adolescents: results from a cross-sectional analysis of the school-based multinational UPRIGHT project.

PURPOSE: The existing evidence suggests that a complete evaluation of mental health should incorporate both psychopathology and mental well-being indicators. However, few studies categorize European adolescents into subgroups based on such complete mental health data. This study used the data on mental well-being and symptoms of mental and behavioral disorders to explore the mental health profiles of adolescents in Europe. METHODS: Data collected from adolescents (N = 3767; mean age 12.4 [SD = 0.9]) from five European countries supplied the information on their mental well-being (personal resilience, school resilience, quality of life, and mental well-being) and mental and behavioral disorder symptoms (anxiety, depression, stress, bullying, cyber-bullying, and use of tobacco, alcohol, or cannabis). Multiple correspondence analysis and cluster analysis were combined to classify the youths into mental health profiles. RESULTS: Adolescents were categorized into three mental health profiles. The "poor mental health" profile (6%) was characterized by low levels of well-being and moderate symptoms of mental disorders. The "good mental health" profile group (26%) showed high well-being and few symptoms of mental disorders, and the "intermediate mental health" profile (68%) was characterized by average well-being and mild-to-moderate symptoms of mental disorders. Groups with higher levels of well-being and fewer symptoms of mental disorders showed lower rates of behavioral problems. Mental well-being indicators strongly contributed to this classification. CONCLUSION: Adolescents with the "intermediate" or "poor" mental health profiles may benefit from interventions to improve mental health. Implications for school-based interventions are discussed. TRIAL REGISTRATION NUMBER (TRN) AND DATE OF REGISTRATION: ClinicalTrials.gov Identifier: NCT03951376. Registered 15 May 2019.

Endothelial Dysfunction: An Intermediate Clinical Feature between Urolithiasis and Cardiovascular Diseases.

An epidemiological relationship between urolithiasis and cardiovascular diseases has extensively been reported. Endothelial dysfunction is an early pathogenic event in cardiovascular diseases and has been associated with oxidative stress and low chronic inflammation in hypertension, coronary heart disease, stroke or the vascular complications of diabetes and obesity. The aim of this study is to summarize the current knowledge about the pathogenic mechanisms of urolithiasis in relation to the development of endothelial dysfunction and cardiovascular morbidities. METHODS: A non-systematic review has been performed mixing the terms "urolithiasis", "kidney stone" or "nephrolithiasis" with "cardiovascular disease", "myocardial infarction", "stroke", or "endothelial dysfunction". RESULTS: Patients with nephrolithiasis develop a higher incidence of cardiovascular disease with a relative risk estimated between 1.20 and 1.24 and also develop a higher vascular disease risk scores. Analyses of subgroups have rendered inconclusive results regarding gender or age. Endothelial dysfunction has also been strongly associated with urolithiasis in clinical studies, although no systemic serum markers of endothelial dysfunction, inflammation or oxidative stress could be clearly related. Analysis of urine composition of lithiasic patients also detected a higher expression of proteins related to cardiovascular disease. Experimental models of hyperoxaluria have also found elevation of serum endothelial dysfunction markers. CONCLUSIONS: Endothelial dysfunction has been strongly associated with urolithiasis and based on the experimental evidence, should be considered as an intermediate and changeable feature between urolithiasis and cardiovascular diseases. Oxidative stress, a key pathogenic factor in the development of endothelial dysfunction has been also pointed out as an important factor of lithogenesis. Special attention must be paid to cardiovascular morbidities associated with urolithiasis in order to take advantage of pleiotropic effects of statins, angiotensin receptor blockers and allopurinol.

Effect of heat treatment on oxalate and hydrocyanic acid levels of malanga corms of two cultivars (Xanthosoma sagittifolium and Colocasia esculenta) in a murine model.

Xanthosoma sagittifolium and Colocasia esculenta contain high levels of nutrients; but have naturally toxic compounds, oxalates and hydrocyanic acid (HCN). The objective of this work was to evaluate the effect of heat treatment on the concentration of antinutrients in malanga corms and its effect on mice. Malanga samples were heated to a boil for 0 to 120 min; oxalates and HCN were determined by spectrophotometry, at 710 and 510 nm, respectively. Pellets were prepared from raw malanga flour (15 and 50%), cooked malanga (15 and 50%) and wheat flour (control) and fed for nine weeks to five groups of six mice each. Cooking of X. sagittifolium corms for 80 min reduced oxalates present by 75% (143 to 35.6 mg/100 g sample), while oxalates in C. esculenta were reduced by 83% (345 to 57.8 mg/100 g sample). HCN levels became negligible after 20 min of cooking. During the nine weeks of feeding the different mice groups showed no significant difference (p > 0.05) between initial and final weight, with respect of the control; mice did not lose their appetite. The results indicate that the consumption of cooked malanga does not pose an evident risk to health, assessed by the reduced level of antinutrients, being an excellent alternative for feeding people in communities with prevalence of food insecurity.

P-sort: an open-source software for cerebellar neurophysiology.

Analysis of electrophysiological data from Purkinje cells (P-cells) of the cerebellum presents unique challenges to spike sorting. Complex spikes have waveforms that vary significantly from one event to the next, raising the problem of misidentification. Even when complex spikes are detected correctly, the simple spikes may belong to a different P-cell, raising the danger of misattribution. To address these identification and attribution problems, we wrote an open-source, semiautomated software called P-sort, and then tested it by analyzing data from P-cells recorded in three species: marmosets, macaques, and mice. Like other sorting software, P-sort relies on nonlinear dimensionality reduction to cluster spikes. However, it also uses the statistical relationship between simple and complex spikes to merge disparate clusters and split a single cluster. In comparison with expert manual curation, occasionally P-sort identified significantly more complex spikes, as well as prevented misattribution of clusters. Three existing automatic sorters performed less well, particularly for identification of complex spikes. To improve the development of analysis tools for the cerebellum, we provide labeled data for 313 recording sessions, as well as statistical characteristics of waveforms and firing patterns of P-cells in three species.NEW & NOTEWORTHY Algorithms that perform spike sorting depend on waveforms to cluster spikes. However, a cerebellar Purkinje-cell produces two types of spikes; simple and complex spikes. A complex spike coincides with the suppression of generating simple spikes. Here, we recorded neurophysiological data from three species and developed a spike analysis software named P-sort that relies on this statistical property to improve both the detection and the attribution of simple and complex spikes in the cerebellum.

From Data to Actions in Intelligent Transportation Systems: A Prescription of Functional Requirements for Model Actionability.

Advances in Data Science permeate every field of Transportation Science and Engineering, resulting in developments in the transportation sector that are data-driven. Nowadays, Intelligent Transportation Systems (ITS) could be arguably approached as a "story" intensively producing and consuming large amounts of data. A diversity of sensing devices densely spread over the infrastructure, vehicles or the travelers' personal devices act as sources of data flows that are eventually fed into software running on automatic devices, actuators or control systems producing, in turn, complex information flows among users, traffic managers, data analysts, traffic modeling scientists, etc. These information flows provide enormous opportunities to improve model development and decision-making. This work aims to describe how data, coming from diverse ITS sources, can be used to learn and adapt data-driven models for efficiently operating ITS assets, systems and processes; in other words, for data-based models to fully become actionable. Grounded in this described data modeling pipeline for ITS, we define the characteristics, engineering requisites and challenges intrinsic to its three compounding stages, namely, data fusion, adaptive learning and model evaluation. We deliberately generalize model learning to be adaptive, since, in the core of our paper is the firm conviction that most learners will have to adapt to the ever-changing phenomenon scenario underlying the majority of ITS applications. Finally, we provide a prospect of current research lines within Data Science that can bring notable advances to data-based ITS modeling, which will eventually bridge the gap towards the practicality and actionability of such models.

Bidirectional short-term plasticity during single-trial learning of cerebellar-driven eyelid movements in mice.

The brain is constantly monitoring its own performance, using error signals to trigger mechanisms of plasticity that help improve future behavior. Indeed, adaptive changes in behavior have been observed after a single error trial in many learning tasks, including cerebellum-dependent eyeblink conditioning. Here, we demonstrate that the plasticity underlying single-trial learning during eyeblink conditioning in mice is bidirectionally regulated by positive and negative prediction errors, has an ephemeral effect on behavior (decays in <1 min), and can be triggered in the absence of errors in performance. We suggest that these three properties of single-trial learning may be particularly useful for driving mechanisms of motor adaptation that can achieve optimal performance in the face of environmental disturbances with a fast timescale.

Pathophysiological Significance of Store-Operated Calcium Entry in Cardiovascular and Skeletal Muscle Disorders and Angiogenesis.

Store-Operated Ca2+ Entry (SOCE) is an important Ca2+ influx pathway expressed by several excitable and non-excitable cell types. SOCE is recognized as relevant signaling pathway not only for physiological process, but also for its involvement in different pathologies. In fact, independent studies demonstrated the implication of essential protein regulating SOCE, such as STIM, Orai and TRPCs, in different pathogenesis and cell disorders, including cardiovascular disease, muscular dystrophies and angiogenesis. Compelling evidence showed that dysregulation in the function and/or expression of isoforms of STIM, Orai or TRPC play pivotal roles in cardiac hypertrophy and heart failure, vascular remodeling and hypertension, skeletal myopathies, and angiogenesis. In this chapter, we summarized the current knowledge concerning the mechanisms underlying abnormal SOCE and its involvement in some diseases, as well as, we discussed the significance of STIM, Orai and TRPC isoforms as possible therapeutic targets for the treatment of angiogenesis, cardiovascular and skeletal muscle diseases.

Lipoxygenase and Its Relationship with Ethylene During Ripening of Genetically Modified Tomato (Solanum lycopersicum).

RESEARCH BACKGROUND: TomloxB is the main isoform of lipoxygenase associated with ripening and senescence of fruits. On the other hand, ethylene, a gaseous hormone, is essential for the regulation of ripening in climacteric fruits like tomatoes. However, the relationship between TomloxB and ethylene production has not been thoroughly studied. Therefore, we aim to assess the effect of exogenous ethylene in transgenic tomatoes that contain a silenced TomloxB gene, and subsequently evaluate lipoxygenase activity, 1-aminocyclopropane-1-carboxylic acid oxidase and ethylene production; as well as to quantify the expression of the genes encoding 1-aminocyclopropane-1-carboxylic acid oxidase and TomloxB. EXPERIMENTAL APPROACH: To investigate the effect of lipoxygenase and 1-aminocyclopropane-1-carboxylic acid oxidase activity, fruits harvested at the stages of break, turning and pink were used. Tomatoes at break stage collected from transgenic and wild type plants were used to determine ethylene production and gene expression. Genetically modified and wild type tomato fruits were exposed to 100 µL/L exogenous ethylene. Lipoxygenase activity was measured spectrophotometrically. Activity of 1-aminocyclopropane-1-carboxylic acid oxidase and ethylene production were determined by gas chromatography. Oligonucleotides for differentially expressed genes: 1-aminocyclopropane-1-carboxylic acid oxidase and TomloxB were used to determine gene expression by real-time PCR. RESULTS AND CONCLUSIONS: The data showed that silencing of TomloxB caused a reduction in lipoxygenase activity and ethylene production in tomato fruits, and also reduced 1-aminocyclopropane-1-carboxylic acid oxidase activity. Hence, the addition of exogenous ethylene increased lipoxygenase activity in all treatments and 1-aminocyclopropane-1-carboxylic acid oxidase activity only in transgenic lines at break stage, consequently there was a positive regulation between TomloxB and ethylene, as increasing the amount of ethylene increased the activity of lipoxygenase. The results suggest that lipoxygenase may be a regulator of 1-aminocyclopropane-1-carboxylic acid oxidase and production of ethylene at break stage. NOVELTY AND SCIENTIFIC CONTRIBUTION: These results lead to a better understanding of the metabolic contribution of TomloxB in fruit ripening and how it is linked to the senescence-related process, which can lead to a longer shelf life of fruits. Understanding this relationship between lipoxygenase and ethylene can be useful for better post-harvest handling of tomatoes.

Comparative transcriptomic analysis unveils interactions between the regulatory CarS protein and light response in Fusarium.

BACKGROUND: The orange pigmentation of the agar cultures of many Fusarium species is due to the production of carotenoids, terpenoid pigments whose synthesis is stimulated by light. The genes of the carotenoid pathway and their regulation have been investigated in detail in Fusarium fujikuroi. In this and other Fusarium species, such as F. oxysporum, deep-pigmented mutants affected in the gene carS, which encodes a protein of the RING-finger family, overproduce carotenoids irrespective of light. The induction of carotenogenesis by light and its deregulation in carS mutants are achieved on the transcription of the structural genes of the pathway. We have carried out global RNA-seq transcriptomics analyses to investigate the relationship between the regulatory role of CarS and the control by light in these fungi. RESULTS: The absence of a functional carS gene or the illumination exert wide effects on the transcriptome of F. fujikuroi, with predominance of genes activated over repressed and a greater functional diversity in the case of genes induced by light. The number of the latter decreases drastically in a carS mutant (1.1% vs. 4.8% in the wild-type), indicating that the deregulation produced by the carS mutation affects the light response of many genes. Moreover, approximately 27% of the genes activated at least 2-fold by light or by the carS mutation are coincident, raising to 40% for an 8-fold activation threshold. As expected, the genes with the highest changes under both regulatory conditions include those involved in carotenoid metabolism. In addition, light and CarS strongly influence the expression of some genes associated with stress responses, including three genes with catalase domains, consistent with roles in the control of oxidative stress. The effects of the CarS mutation or light in the transcriptome of F. oxysporum were partially coincident with those of F. fujikuroi, indicating the conservation of the objectives of their regulatory mechanisms. CONCLUSIONS: The CarS RING finger protein down-regulates many genes whose expression is up-regulated by light in wild strains of the two investigated Fusarium species, indicating a regulatory interplay between the mechanism of action of the CarS protein and the control by light.

UPRIGHT, a resilience-based intervention to promote mental well-being in schools: study rationale and methodology for a European randomized controlled trial.

BACKGROUND: Adolescence is crucial period for laying the foundations for healthy development and mental well-being. The increasing prevalence of mental disorders amongst adolescents makes promotion of mental well-being and prevention interventions at schools important. UPRIGHT (Universal Preventive Resilience Intervention Globally implemented in schools to improve and promote mental Health for Teenagers) is designed as a whole school approach (school community, students and families) to promote a culture of mental well-being and prevent mental disorders by enhancing resilience capacities. The present article aims at describing the rationale, conceptual framework, as well as methodology of implementation and evaluation of the UPRIGHT intervention. METHODS: UPRIGHT project is a research and innovation project funded by the European Union's Horizon 2020 Research and Innovation programme under grant agreement No. 754919 (Duration: 48 months). The theoretical framework has been developed by an innovative and multidisciplinary approach using a co-creation process inside the UPRIGHT Consortium (involving seven institutions from Spain, Italy, Poland, Norway, Denmark, and Iceland). Resulted is the UPRIGHT programme with 18 skills related to 4 components: Mindfulness, Coping, Efficacy and Social and Emotional Learning. Among the five Pan-European regions, 34 schools have been currently involved (17 control; 17 intervention) and around 6000 adolescents and their families are foreseen to participate along a 3-year period of evaluation. Effectiveness of the intervention will be evaluated as a randomized controlled trial including quantitative and qualitative analysis in the five Pan-European regions representative of the cultural and socioeconomic diversity. The cost-effectiveness assessment will be performed by simulation modelling methods. DISCUSSION: We expect a short- to medium-term improvement of mental well-being in adolescents by enhancing resilience capacities. The study may provide robust evidence on intrapersonal, familiar and social environmental resilience factors promoting positive mental well-being. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03951376 . Registered 15 May 2019.

Data Stream Mining Applied to Maximum Wind Forecasting in the Canary Islands.

The Canary Islands are a well known tourist destination with generally stable and clement weather conditions. However, occasionally extreme weather conditions occur, which although very unusual, may cause severe damage to the local economy. The ViMetRi-MAC EU funded project has among its goals, managing climate-change-associated risks. The Spanish National Meteorology Agency (AEMET) has a network of weather stations across the eight Canary Islands. Using data from those stations, we propose a novel methodology for the prediction of maximum wind speed in order to trigger an early alert for extreme weather conditions. The methodology proposed has the added value of using an innovative kind of machine learning that is based on the data stream mining paradigm. This type of machine learning system relies on two important features: models are learned incrementally and adaptively. That means the learner tunes the models gradually and endlessly as new observations are received and also modifies it when there is concept drift (statistical instability), in the modeled phenomenon. The results presented seem to prove that this data stream mining approach is a good fit for this kind of problem, clearly improving the results obtained with the accumulative non-adaptive version of the methodology.

Preliminary results from the use of intraoperative real-time biliary oxygen monitoring in liver transplantation.

BACKGROUND AND AIMS: Biliary anastomosis is a frequent area of complications after liver transplantation (LT) and a potential area of "microangiopathy". The concept of a "marginal bile duct" is unexplored. The main aim was to make a preliminary evaluation of the utility of an innovative real-time oxygen microtension (pO2mt) testing device for the assessment of bile duct viability during LT and to correlate these pO2mt values with microvascular tissue quality by histopathology and outcomes. PATIENTS AND METHODS: Observational prospective cohort study with 23 patients. Oxygen microtension measurements were made placing a micropO2 probe in different areas of recipient and donor's bile duct intraoperative. RESULTS: Mean pO2mt in the graft bile duct at the level of the anastomosis 103.82 (31-157) mm Hg, being 121.52 (55-174) mm Hg 1.5 cm proximal to the hilar plate (P < 0.001). Mean pO2mt in the recipient's bile duct was 117.87 (62-185) mm Hg, while a value of 137.30 (81-198) mm Hg was observed 1.5 cm distal to the anastomosis (P < 0.001). Cystic duct resection (12 cases) was also related with higher pO2mt values at anastomosis [117.8 (93-157) vs 88.54 (31-124) mm Hg] and distal to anastomosis [135.6 (111-174) vs 106.2 (55-133) mm Hg; P < 0.001]. Patients with 1-, 3-, and 12-month biliary complications had significantly lower pO2mt in the intraoperative measurements. CONCLUSION: Our preliminary results show that distal borders of donor and recipient bile ducts may be low-vascularized areas. Tissue pO2mt is significantly higher in areas close to the hilar plate and to the duodenum in donor and recipient's sides, respectively. Bile duct injury and biliary complications are associated with worse tissue pO2mt.

Using Ontologies for the Online Recognition of Activities of Daily Living.

The recognition of activities of daily living is an important research area of interest in recent years. The process of activity recognition aims to recognize the actions of one or more people in a smart environment, in which a set of sensors has been deployed. Usually, all the events produced during each activity are taken into account to develop the classification models. However, the instant in which an activity started is unknown in a real environment. Therefore, only the most recent events are usually used. In this paper, we use statistics to determine the most appropriate length of that interval for each type of activity. In addition, we use ontologies to automatically generate features that serve as the input for the supervised learning algorithms that produce the classification model. The features are formed by combining the entities in the ontology, such as concepts and properties. The results obtained show a significant increase in the accuracy of the classification models generated with respect to the classical approach, in which only the state of the sensors is taken into account. Moreover, the results obtained in a simulation of a real environment under an event-based segmentation also show an improvement in most activities.

Protein Activity of the Fusarium fujikuroi Rhodopsins CarO and OpsA and Their Relation to Fungus-Plant Interaction.

Fungi possess diverse photosensory proteins that allow them to perceive different light wavelengths and to adapt to changing light conditions in their environment. The biological and physiological roles of the green light-sensing rhodopsins in fungi are not yet resolved. The rice plant pathogen Fusarium fujikuroi exhibits two different rhodopsins, CarO and OpsA. CarO was previously characterized as a light-driven proton pump. We further analyzed the pumping behavior of CarO by patch-clamp experiments. Our data show that CarO pumping activity is strongly augmented in the presence of the plant hormone indole-3-acetic acid and in sodium acetate, in a dose-dependent manner under slightly acidic conditions. By contrast, under these and other tested conditions, the Neurospora rhodopsin (NR)-like rhodopsin OpsA did not exhibit any pump activity. Basic local alignment search tool (BLAST) searches in the genomes of ascomycetes revealed the occurrence of rhodopsin-encoding genes mainly in phyto-associated or phytopathogenic fungi, suggesting a possible correlation of the presence of rhodopsins with fungal ecology. In accordance, rice plants infected with a CarO-deficient F. fujikuroi strain showed more severe bakanae symptoms than the reference strain, indicating a potential role of the CarO rhodopsin in the regulation of plant infection by this fungus.

Orai1 and TRPC1 Proteins Co-localize with CaV1.2 Channels to Form a Signal Complex in Vascular Smooth Muscle Cells.

Voltage-dependent CaV1.2 L-type Ca2+ channels (LTCC) are the main route for calcium entry in vascular smooth muscle cells (VSMC). Several studies have also determined the relevant role of store-operated Ca2+ channels (SOCC) in vascular tone regulation. Nevertheless, the role of Orai1- and TRPC1-dependent SOCC in vascular tone regulation and their possible interaction with CaV1.2 are still unknown. The current study sought to characterize the co-activation of SOCC and LTCC upon stimulation by agonists, and to determine the possible crosstalk between Orai1, TRPC1, and CaV1.2. Aorta rings and isolated VSMC obtained from wild type or smooth muscle-selective conditional CaV1.2 knock-out (CaV1.2KO) mice were used to study vascular contractility, intracellular Ca2+ mobilization, and distribution of ion channels. We found that serotonin (5-HT) or store depletion with thapsigargin (TG) enhanced intracellular free Ca2+ concentration ([Ca2+]i) and stimulated aorta contraction. These responses were sensitive to LTCC and SOCC inhibitors. Also, 5-HT- and TG-induced responses were significantly attenuated in CaV1.2KO mice. Furthermore, hyperpolarization induced with cromakalim or valinomycin significantly reduced both 5-HT and TG responses, whereas these responses were enhanced with LTCC agonist Bay-K-8644. Interestingly, in situ proximity ligation assay revealed that CaV1.2 interacts with Orai1 and TRPC1 in untreated VSMC. These interactions enhanced significantly after stimulation of cells with 5-HT and TG. Therefore, these data indicate for the first time a functional interaction between Orai1, TRPC1, and CaV1.2 channels in VSMC, confirming that upon agonist stimulation, vessel contraction involves Ca2+ entry due to co-activation of Orai1- and TRPC1-dependent SOCC and LTCC.

Increased Postharvest Life of TomLox B Silenced Mutants of Tomato (Solanum lycopersicum) Var. TA234.

A healthy lifestyle includes fruits and vegetables consumption. Tomato is one of the most consumed vegetables, although it is susceptible to physical damage through postharvest handling, thus leading to important losses. Softening is an important variable during tomato ripening; excessive softening is undesirable and leads to postharvest losses. TomloxB plays an important role in ripening, mainly in the loss of cellular integrity caused by fatty acids released from the lipid matrix of membranes that initiate oxidative deterioration, which is in turn carried into senescence. In order to increase postharvest life, we produced transgenic tomato plants via Rhizobium radiobacter with tomato lipoxygenase B (TomloxB) antisense constructs under control of the cauliflower mosaic virus (CaMV) 35S promoter. Lipoxygenase activity and firmness were measured in tomato fruit and the fatty acids profile was determined. Transgenic fruits were maintained for 40 days at room temperature in optimal conditions, whereas wild type fruits remained in similar conditions for only six days. Firmness in pink and red stages was significantly lower in wild type fruits than in two transgenic lines. Linolenic acid was the most important fatty acid consumed by lipoxygenase in both turning and pink stages of ripening. Lipoxygenase activity was smaller in transformed fruits in comparison with the wild type. These results suggest that silencing the TomloxB gene promoted significant changes in the physiology of transformed tomatoes, being the increase in postharvest life the most important.

Phospholipase A2 as a Molecular Determinant of Store-Operated Calcium Entry.

Activation of phospholipases A2 (PLA2) leads to the generation of biologically active lipid products that can affect numerous cellular events. Ca(2+)-independent PLA2 (iPLA2), also called group VI phospholipase A2, is one of the main types forming the superfamily of PLA2. Beside of its role in phospholipid remodeling, iPLA2 has been involved in intracellular Ca(2+) homeostasis regulation. Several studies proposed iPLA2 as an essential molecular player of store operated Ca(2+) entry (SOCE) in a large number of excitable and non-excitable cells. iPLA2 activation releases lysophosphatidyl products, which were suggested as agonists of store operated calcium channels (SOCC) and other TRP channels. Herein, we will review the important role of iPLA2 on the intracellular Ca(2+) handling focusing on its role in SOCE regulation and its implication in physiological and/or pathological processes.

Epidemiology of Barrett's esophagus and esophageal adenocarcinoma in Spain. A unicentric study.

INTRODUCTION: Barrett's esophagus (BE) is an acquired disease defined by the presence of intestinal metaplasia with goblet cells in the distal esophagus. The prevalence of BE has increased dramatically over the last years. AIMS: The primary aims of the study were to analyze the characteristics of BE and esophageal adenocarcinoma (EAC) in a Spanish health district during a follow-up period. METHODOLOGY: Sociodemographic factors, alcohol consumption and cigarette smoking were analyzed. We also studied the histological behavior and cause of death in each group. RESULTS: In the present study 430 patients were included, 338 with BE and 92 with EAC. Incidence rates have risen from 2.25 and 1.25 per 100,000 inhabitants in 1996 to 6.5 and 4.75 per 100,000 in 2011, respectively. In the EAC group, male gender, age and alcohol consumption were higher in comparison to the BE group, and the overall survival was 23 months. In the BE group, the main causes of death were non-esophageal cancer and cardiovascular disease. CONCLUSIONS: The incidence and prevalence rates of AEC and BE have risen over the past years. Risk factors for these conditions were male gender, age and alcohol consumption. Long BE (> 3 cm) is involved in dysplasia progression. AEC diagnosis mainly occurs after neoplasia is detected and, in a few cases, due to a previous BE. Cardiovascular diseases and non-esophageal cancers have been found to be the main cause of death in BE patients.