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Black carrot (Daucus carota ssp. sativus var. atrorubens Alef.) is widely recognized for its bioactive compounds and antioxidant properties. The black carrot of Cuevas Bajas (Málaga) is a local variety characterized by a black/purple core, which differs from other black carrot varieties. Therefore, this autochthonous variety was characterized according to the root size and the harvesting season by means of a study of its antioxidant capacity analyzed by three methods, its total carotenoids content, and its sugars and phenolic compounds profile by ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-MS). A total of 20 polyphenolic compounds were quantified in 144 samples analyzed. The anthocyanidins group was observed to be the most abundant, followed by the hydroxycinnamic acids group. Moreover, pelargonidin 3-sambubioside was observed in black carrot for the first time. The medium-sized carrots presented the highest content of phenolic compounds, largely due to their significantly higher anthocyanidins content. Comparatively, the small carrots showed a higher content of simple sugars than the large ones. Regarding the influence of season, significantly higher quantities of glucose and fructose were observed in the late-season carrots, while sucrose was the main sugar in early-season samples. No significant differences were observed in the total carotenoid content of black carrot.

The global demand for organic food products has rapidly increased over the last years, becoming an emerging niche market targeting the high-income segment. The higher retailing price for organic food products may increase the risk of fraudulent practices at the different stages of the food supply chain, and consequently, substantial control is needed. Currently, the authentication of organic food products, such as those of plant origin, remains a key challenge in analytical chemistry. While stable isotopes have emerged as a powerful tool for this purpose, most studies have focused on crops, missing the agricultural inputs used for fertilization that influence the isotopic values of the crops. In this study, we aimed to isotopically characterize commonly used fertilizers, soil conditioners, and micronutrient fertilizers in intensive organic agriculture in the largest organic production region in the world (Almería, Spain). Our goal was to clarify the limitations that nitrogen isotopic fingerprinting presents for the fertilizer input industry and to characterize the organic inputs. The conventional fertilizers analyzed in this study showed low δ15N values compared to their organic counterparts, except for some plant-based fertilizers, protein hydrolyzed fertilizers, and chelated nutrients. Both protein hydrolyzed fertilizers and micronutrient fertilizers presented a wide range of variability in their δ15N values, including some very low or even negative values, more similar to those of conventional fertilizers. The results of this study highlight the challenges of authenticating organic foods in agriculture when using nitrogen isotope analysis.

Existing direct and inverse kinematic models of planar parallel robots assume that the robot’s active joints are all at the bases. However, this approach becomes excessively complex when modeling a planar parallel robot in which the active joints are within one single kinematic chain. To address this problem, our article unveils an alternative for a 3RRR symmetric planar robot modeling technique for the derivation of the robot workspace and the analysis of its direct and inverse kinematics. The workspace was defined using a system of inequalities, and the direct and inverse kinematics models were generated using vectorial analysis and an optimized geometrical approach, respectively. The resulting models are systematically presented and validated. Two final model renditions are delivered supplying a thorough equation analysis and an applicability discussion based on the importance of the robot’s mobile platform orientation. The advantages of this model are discussed in comparison to the traditional modeling approach: whereas conventional techniques require the solution of complex eighth-degree polynomials for the analysis of the active joint configuration of these robots, these models provide an efficient back-of-the-envelope analysis approach that requires the solution of a simple second-degree polynomial.

Currently, immunotherapy based on PD-1/PD-L1 pathway blockade has improved survival of non-small cell lung cancer (NSCLC) patients. However, differential responses have been observed by sex, where men appear to respond better than women. Additionally, adverse effects of immunotherapy are mainly observed in women. Studies in some types of hormone-dependent cancer have revealed a role of sex hormones in anti-tumor response, tumor microenvironment and immune evasion. Estrogens mainly promote immune tolerance regulating T-cell function and modifying tumor microenvironment, while androgens attenuate anti-tumor immune responses. The precise mechanism by which sex and sex hormones may modulate immune response to tumor, modify PD-L1 expression in cancer cells and promote immune escape in NSCLC is still unclear, but current data show how sexual differences affect immune therapy response and prognosis. This review provides update information regarding anti-PD-1/PD-L immunotherapeutic efficacy in NSCLC by sex, analyzing potential roles for sex hormones on PD-L1 expression, and discussing a plausible of sex and sex hormones as predictive response factors to immunotherapy.

Mobile robotics has been used in recent years to provide various types of services in fields such as agriculture, surveillance, rehabilitation, space exploration, and logistics, among others. In many cases, mobile robots need to overcome complex obstacles where traditional wheels are not the best solution, and many researchers have proposed legged wheel hybrid designs. This paper presents a comprehensive study on the effect of the geometry of legged wheels on the performance of mobile robots in climbing stairs. The method used to develop this research is dynamic simulation, in which the parameters that affect the kinematics and dynamics of the robot are included. Subsequently, the results of how the robot manages to perform the test, the torque of the motors, and the contact force of the wheels are analyzed. The main hypothesis of this research is that the opening of the legs of the wheels is a geometric parameter that determines whether the structure will be able to climb the stairs. After 63 simulations, the proportional relationship between the diagonal of the stands and the opening of the wheel legs ranges between 1.11 and 1.53. This parameter showed a strong correlation with the torque of the motors and significant differences in terms of the simulations that succeeded in climbing the stairs and those that did not. These results were used to state a design method for flat, robotic structures using legged wheels. This method was validated by an additional simulation that was performed for a four-legged wheel. It can be concluded that the contribution of this work is a series of steps with which to design these mechanical structures to climb the stairs based on the proposed indicator.

Owing to their ease of implementation, proportional-integral-derivative (PID) control systems are widely used to control physical systems. However, when environmental disturbances or changes in system parameters occur, the complexity of tuning the gains of PID controllers increases because, in these cases, their performance decreases. To solve this problem, an AI-based online self-tuning algorithm adjusts the PID gains when system parameters are changed. Thus, the objective of this study was to develop a PID control algorithm with adaptive parameter tuning using artificial hydrocarbon networks, which is a supervised learning artificial intelligence technique inspired by hydrocarbon networks. Artificial hydrocarbon networks were designed to be trained using a prior set of data. Therefore, the motivation of this study was to demonstrate that they can be trained in real time for use in the control of nonlinear systems. Because this type of network has not been commonly used for this specific application, existing studies on adaptive control based on artificial neural networks were taken as a reference. The AMSGrad optimization algorithm was used to train the parameters in real time, for which a “continuous” model of artificial hydrocarbon networks was also proposed. Finally, an algorithm capable of adapting to variations in the operating conditions of a tank system was successfully designed, although its performance was similar to that of the Ziegler-Nichols method.

Abstract BIRC3 is monoallelically deleted in up to 80% of chronic lymphocytic leukemia (CLL) cases harboring del(11q). In addition, truncating mutations in the remaining allele of this gene can lead to BIRC3 biallelic inactivation, which has been shown to be a marker for reduced survival in CLL. Nevertheless, the biological mechanisms by which these lesions could contribute to del(11q) CLL pathogenesis and progression are partially unexplored. We implemented the CRISPR/Cas9-editing system to generate isogenic CLL cell lines harboring del(11q) and/or BIRC3 mutations, modeling monoallelic and biallelic BIRC3 loss. Our results reveal that monoallelic BIRC3 deletion in del(11q) cells promotes non-canonical NF-κB signaling activation via RelB-p52 nuclear translocation, being these effects allelic dose-dependent and therefore further enhanced in del(11q) cells with biallelic BIRC3 loss. Moreover, we demonstrate ex vivo in primary cells that del(11q) cases including BIRC3 within their deleted region show evidence of non-canonical NF-κB activation which correlates with high BCL2 levels and enhanced sensitivity to venetoclax. Furthermore, our results show that BIRC3 mutations in del(11q) cells promote clonal advantage in vitro and accelerate leukemic progression in an in vivo xenograft model. Altogether, this work highlights the biological bases underlying disease progression of del(11q) CLL patients harboring BIRC3 deletion and mutation.

The aim of this study was to evaluate the impact of in vitro gastrointestinal digestion and colonic fermentation on the polyphenol compounds from different varieties of pistachio by UHPLC-HRMS analysis. The total polyphenol content decreased significantly, mostly during oral (recoveries of 27 to 50%) and gastric digestion (recoveries of 10 to 18%), with no significant changes after the intestinal phase. After in vitro digestion, the hydroxybenzoic acids and the flavan-3-ols were the main compounds found in pistachio, with respective total polyphenol contents of 73 to 78% and 6 to 11%. More specifically, the main compounds determined after in vitro digestion were 3,4,5-trihydroxybenzoic acid, vanillic hexoside and epigallocatechin gallate. The colonic fermentation affected the total phenolic content of the six varieties studied, with a recovery range of 11 to 25% after 24 h of fecal incubation. A total of twelve catabolites were identified after fecal fermentation, the main compounds being the 3-(3′-hydroxyphenyl)propanoic, 3-(4′-hydroxyphenyl)propanoic, 3-(3′,4′-dihydroxyphenyl)propanoic, 3-hydroxyphenylacetic acids and 3,4-dihydroxyphenyl-ɣ-valerolactone. Based on these data, a catabolic pathway for colonic microbial degradation of phenolic compounds is proposed. The catabolites identified at the end of the process are potentially responsible for the health properties attributed to pistachio consumption.

Renal vasculature, which is highly innervated by sympathetic fibers, contributes to cardiovascular homeostasis. This renal sympathetic outflow is inhibited by 5-HT in normoglycaemic rats. Considering that diabetes induces cardiovascular complications, we aimed to determine whether diabetic state modifies noradrenergic input at renal level and its serotonergic modulation in rats. Alloxan diabetic rats were anaesthetized (pentobarbital; 60 mg/kg i.p.) and prepared for in situ autoperfusion of the left kidney to continuously measure systemic blood pressure (SBP), heart rate (HR), and renal perfusion pressure (RPP). Electrical stimulation of renal sympathetic outflow induces frequency-dependent increases (Δ) in RPP (23.9 ± 2.1, 59.5 ± 1.9, and 80.5 ± 3.5 mm Hg at 2, 4, and 6 Hz, respectively), which were higher than in normoglycaemic rats, without modifying HR or SBP. Intraarterial bolus of 5-HT and 5-CT (5-HT1/5/7 agonist) reduced electrically induced ΔRPP. Only L-694,247 (5-HT1D agonist) reproduced 5-CT inhibition on sympathetic-induced vasoconstrictions, whereas it did not modify exogenous noradrenaline-induced ΔRPP. 5-CT inhibition was exclusively abolished by i.v. bolus of LY310762 (5-HT1D antagonist). An inhibitor of guanylyl cyclase, ODQ (i.v.), completely reversed the L-694,247 inhibitory effect. In conclusion, diabetes induces an enhancement in sympathetic-induced vasopressor responses at the renal level. Prejunctional 5-HT1D receptors, via the nitric oxide pathway, inhibit noradrenergic-induced vasoconstrictions in diabetic rats.

In this study, in vitro cultures and micropropagated plants of two Mediterranean aromatic plants, Lavandula viridis L’Hér and Thymus lotocephalus López and Morales, were exposed to different temperatures (15, 20, 25, and 30 °C). The effect of temperature on the levels of hydrogen peroxide (H2O2), lipid peroxidation, and osmoprotectants (proline, soluble sugars, and soluble proteins), as well as on the phenolic profile by HPLC-HRMS and intermediates of the secondary metabolism (phenylalanine ammonia lyase (PAL) activity and shikimic acid content), was investigated. Moreover, the antioxidant activity of the plant extracts was also analyzed. Overall, considering the lipid peroxidation and H2O2 content, the extreme temperatures (15 and 30 °C) caused the greatest damage to both species, but the osmoprotectant response was different depending on the species and plant material. In both species, phenolic compounds and related antioxidant activity increased with the rise in temperature in the micropropagated plants, while the opposite occurred in in vitro cultures. L. viridis cultures showed the highest biosynthesis of rosmarinic acid (92.6 g/kgDW) at 15 °C and seem to be a good alternative to produce this valuable compound. We conclude that contrasting temperatures greatly influence both species’ primary and secondary metabolism, but the response is different depending on the plant micropropagation stage.

Vanadium (V) toxicity depends on its oxidation state; it seems that vanadium pentoxide (V2O5) is the most toxic to the living cells. It has been reported that oral administration induces changes in motor activity and learning; in rats, I.P. administration increases lipid peroxidation levels in the cerebellum and the concentration of free radicals in the hippocampus and cerebellum. Mice that inhaled V2O5 presented a reduced number of tubulin+ in Leydig and Sertoli cells; it has also been reported that inhaled V2O5 induces loss of dendritic spines, necrosis, and hippocampus neuropil alterations; considering the direct consequence of the interaction of V with cytoskeletal components, makes us believe that V2O5 exposure could cause neuronal death in the hippocampus similar to that seen in Alzheimer disease. This work aimed to determine pyramidal hippocampal CA1 cytoskeletal alterations with Bielschowsky stain in rats exposed to V2O5. Male Wistar rats inhaled 0.02 M of V2O5 one h two times a week for two and six months. We found that rats, which inhaled V2O5 reached 56,57% of dead neurons after six months of inhalation; we recognize strong argyrophilic and collapsed somas and typical flame-shaped in all V-exposed rats hippocampus CA1 compared to controls. We also observe somatodendritic distortions. Axons and dendrites displayed thick dark bands replaced by noticeable thickening and nodosities and the cytoskeleton fibrillary proteins' linear traces. Our findings suggest that V2O5 inhalation induces Alzheimer-like cell death with evident cytoskeletal alterations.

Owing to their compliance with most shapes, soft actuators are regarded as cost-effective solutions for grasping irregular objects. The material properties of nonlinear elastic polymers are considered necessary for the correct implementation of these actuators. The analysis tends to be complex even for simple movements defined by theoretically infinite degrees of freedom. This study offers a mathematical model that outlines a relationship between the energy provided by a pressure source and the expected behavior of multi-chamber pneumatic soft actuators through hyper-elastic material deformation interpretation, geometric approximations, and the vectorial representations of their segments. Digitally analyzed empirical results measured through lateral pictures of an actuator were taken at different pressure references. Direct comparisons between the average value of the tested angles and those calculated through the tuned mathematical model provide a maximum error of 0.647° for small deformations and an improved accuracy at higher pressure inputs. This study offers a valid tool applicable to the design of soft actuators and their further analysis without the need for overly complex methods.

The impact of drought stress induced by polyethylene glycol (PEG) on morphological, physiological, (bio)chemical, and biological characteristics of Thymus lotocephalus López and Morales shoot cultures have been investigated, as well as the potential of iron oxide nanoparticles, salicylic acid, and methyl jasmonate (MeJA) as alleviating drought stress agents. Results showed that PEG caused oxidative stress in a dose-dependent manner, raising H2O2 levels and reducing shoots’ growth, photosynthetic pigment contents, and phenolic compounds production, especially phenolic acids, including the major compound rosmarinic acid. Moreover, Fourier Transform Infrared Spectra analysis revealed that PEG treatment caused changes in shoots’ composition, enhancing terpenoids biosynthesis. PEG also decreased the biological activities (antioxidant, anti-tyrosinase, and photoprotective) of the eco-friendly extracts obtained with a Natural Deep Eutectic Solvent. MeJA was the most efficient agent in protecting cells from oxidative damage caused by drought, by improving the biosynthesis of phenolics, like methyl 6-O-galloyl-β-D-glucopyranoside and salvianolic acids, as well as improving the extracts’ antioxidant activity. Altogether, the obtained results demonstrated a negative impact of PEG on T. lotocephalus shoots and an effective role of MeJA as a mitigating agent of drought stress. Additionally, extracts showed a good potential to be used in the cosmetics industry as skincare products.

Accumulation of misfolded proteins in the brain is a common hallmark of most age-related neurodegenerative diseases. Previous studies from our group identified the presence of anti-inflammatory and antioxidant compounds in leaves derived from the Chilean berry Ugni molinae (murtilla), in addition to show a potent anti-aggregation activity in models of Alzheimer´s disease. However, possible beneficial effects of berry extracts of murtilla was not investigated. Here we evaluated the efficacy of fruit extracts from different genotypes of Chilean-native U. molinae on reducing protein aggregation using cellular models of Huntington´s disease and assess the correlation with their chemical composition. Berry extraction was performed by exhaustive maceration with increasing-polarity solvents. An unbiased automatic microscopy platform was used for cytotoxicity and protein aggregation studies in HEK293 cells using polyglutamine-EGFP fusion proteins, followed by secondary validation using biochemical assays. Phenolic-rich extracts from murtilla berries of the 19-1 genotype (ETE 19-1) significantly reduced polyglutamine peptide aggregation levels, correlating with the modulation in the expression levels of autophagy-related proteins. Using LC-MS and molecular network analysis we correlated the presence of flavonoids, phenolic acids, and ellagitannins with the protective effects of ETE 19-1 effects on protein aggregation. Overall, our results indicate the presence of bioactive components in ethanolic extracts from U. molinae berries that reduce the load of protein aggregates in living cells.

Pistachio (Pistacia vera L.) is a nut with a good adaptability to the Mediterranean conditions of cultivation, specifically in the Andalusian region, becoming an emerging crop. Moreover, it has been getting attention in the past years for the great content of bioactive compounds such as polyphenols. Although some studies have reported the polyphenolic profile of pistachios, most of them have analyzed the hull part, considered as a residue, and not the kernel which is the edible part. Therefore, characterization of eleven varieties of pistachios kernels cultivated in Andalusia and harvested in 2019 and 2020 was carried out by UHPLC-MS (ultra-high-performance liquid chromatography high-resolution mass spectrometry). The identification and quantification of 56 polyphenolic compounds was performed, being the hydroxybenzoic acids group the most abundant with a 71–86% of the total amount followed by flavan-3-ols group that accounted for 8–24%. Moreover, 3,4-dihydroxybenzoic acid was the main compound in most of the varieties, followed by vanillic acid hexoside. Larnaka, Avdat, Aegina, and Mateur presented the highest amount of total polyphenols, while Kalehghouchi, Joley, Lost Hills, Kerman, and Golden Hills were the varieties with the lowest content. Regarding the harvest season, no significant differences (p < 0.01) were found in the total amount of polyphenols between 2019 and 2020. In addition, the antioxidant activity was measured by DPPH (1,1-diphenyl-2-picryl-hydrazyl), ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), and ORAC (oxygen radical absorbance capacity) assays, showing a similar trend as that of the polyphenols.

El presente trabajo describe la experiencia de integración tecnológica en el caso de estudio de Studer Innotec, compañía que fabrica inversores/cargadores y reguladores en Suiza, utilizando un inversor/cargador se rediseño el sistema de transporte mejorando la eficiencia de la cadena agro-productiva de la post cosecha del banano, desarrollado en el sector atlántico de Honduras. Actualmente el sistema de transporte de banano se realiza en cable vía utilizando animales de carga (mulas) haciendo un proceso lento, además de generar costos por el cuido de estos animales o por máquinas subidas al cable vía que utilizan un motor de combustión diésel acoplado a una bomba de aceite, los cuales generan emisiones de gases que contaminan el ambiente e incluso el mismo banano. Con el objetivo de integrar equipos industriales que permitan recolectar una mayor cantidad de racimos que en el sistema actual, asegurando las propiedades de calidad del banano se presenta el rediseño de una maquina utilizadas para el transporte del banano, cambiando su motor diésel de 9 hp, por un motor trifásico de 5 hp, donde el mayor reto ha sido pasar la energía eléctrica almacenada en baterías por un inversor, hasta llegar a un variador de frecuencia, con el que se controló el torque y velocidad del motor, permitiendo un mejor uso del equipo con lo que se ha logrado transportar más de 70 racimos de banano en un recorrido mayor a 800 metros dentro de la finca.

A data-processing and statistical analysis workflow was proposed to evaluate the metabolic changes and its contribution to the sensory characteristics of different wines. This workflow was applied to rosé wines from different fermentation strategies. The metabolome was acquired by means of two high-throughput techniques: gas chromatography–mass spectrometry (GC-MS) and liquid chromatography–mass spectrometry (LC-MS) for volatile and non-volatile metabolites, respectively, in an untargeted approach, while the sensory evaluation of the wines was performed by a trained panel. Wine volatile and non-volatile metabolites modulation was independently evaluated by means of partial least squares discriminant analysis (PLS-DA), obtaining potential markers of the fermentation strategies. Then, the complete metabolome was integrated by means of sparse generalised canonical correlation analysis discriminant analysis (sGCC-DA). This integrative approach revealed a high link between the volatile and non-volatile data, and additional potential metabolite markers of the fermentation strategies were found. Subsequently, the evaluation of the contribution of metabolome to the sensory characteristics of wines was carried out. First, the all-relevant metabolites affected by the different fermentation processes were selected using PLS-DA and random forest (RF). Each set of volatile and non-volatile metabolites selected was then related to the sensory attributes of the wines by means of partial least squares regression (PLSR). Finally, the relationships among the three datasets were complementary evaluated using regularised generalised canonical correlation analysis (RGCCA), revealing new correlations among metabolites and sensory data.

The present study aimed at evaluating the effectiveness of different natural deep eutectic solvents (NADES) on the extraction of phenolic compounds from Lavandula pedunculata subsp. lusitanica (Chaytor) Franco, on the antioxidant activity, and acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and tyrosinase (Tyr) inhibitory capacities. Ten different NADES were used in this research and compared with conventional solvents. Ultrasound-assisted extraction (UAE) for 60 min proved to be the best extraction condition, and proline:lactic acid (1:1) and choline chloride:urea (1:2) extracts showed the highest total phenolic contents (56.00 ± 0.77 mgGAE/gdw) and antioxidant activity [64.35 ± 1.74 mgTE/gdw and 72.13 ± 0.97 mgTE/gdw in 2.2-diphenyl-1-picrylhydrazyl (DPPH) and 2.2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) methods, respectively]. These extracts also exhibited enzymes inhibitory capacity particularly against Tyr and AChE. Even so, organic acid-based NADES showed to be the best extractants producing extracts with considerable ability to inhibit enzymes. Twenty-four phenolic compounds were identified by HPLC-HRMS, being rosmarinic acid, ferulic acid and salvianolic acid B the major compounds. The results confirmed that the combination of UAE and NADES provide an excellent alternative to organic solvents for sustainable and green extraction, and have huge potential for use in industrial applications involving the extraction of bioactive compounds from plants.

This study aims to determine the changes in, and bioaccessibility of, polyphenols and organosulfur compounds (OSCs) during the simulated gastrointestinal digestion of black onion, a novel product derived from fresh onion by a combination of heat and humidity treatment, and to compare it with its fresh counterpart. Fresh and black onions were subjected to in-vitro gastrointestinal digestion, and their polyphenol and OSC profiles were determined by ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS). Although to a lesser extent than in the fresh onion, the phenolic compounds in the black variety remained stable during the digestion process, presenting a higher bioaccessibility index (BI) with recovery corresponding to 41.1%, compared with that of fresh onion (23.5%). As for OSCs, apart from being more stable after the digestion process, with a BI of 83%, significantly higher quantities (21 times higher) were found in black onion than in fresh onion, suggesting that the black onion production process has a positive effect on the OSC content. Gallic acid, quercetin, isorhamnetin, and ɣ-glutamyl-S-(1-propenyl)-L-cysteine sulfoxide were the most bioaccessible compounds in fresh onion, while isorhamnetin, quercetin-diglucoside, ɣ-glutamyl-S-methyl-L-cysteine sulfoxide and methionine sulfoxide were found in black onion. These results indicate that OSCs and polyphenols are more bioaccessible in black onion than in fresh onion, indicating a positive effect of the processing treatment.

Black carrot has been attracting increasing thanks to its high bioactive compound content. This study presents the polyphenol bio-accessibility of black carrot and two derived products (black carrot snack (BC snack) and black carrot seasoning (BC seasoning)) after in vitro gastrointestinal digestion and colonic fermentation. Additionally, antioxidant activity was measured by 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) assays. Nine flavonoids and eight anthocyanins were determined by ultra high-performance liquid chromatography high resolution mass spectrometry (UHPLC-HRMS) analysis, the predominant compounds being the hydroxycinnamic acids 3-O-feruloylquinic acid, 4-O-feruloylquinic acid and chlorogenic acid. The BC snack (108 µmol/g DW) presented the highest total polyphenol content, followed by BC seasoning (53 µmol/g DW) and black carrot (11.4 µmol/g DW). The main polyphenols still bio-accessible after in vitro digestion were the hydroxycinnamic acids, with mean recovery rates of 113 % for black carrot, 69% for BC snack and 81% for BC seasoning. The incubation of black carrot and its derived products with human faecal bacterial resulted in the complete degradation of anthocyanins and in the formation of mainly 3-(4′-hydroxyphenyl)propanoic acid as the major catabolic event. In conclusion, our results suggest that the black carrot matrix impacts significantly affects the bio-accessibility of polyphenols and, therefore, their potential health benefits.

Mango (Mangifera indica L.), a fruit with sensorial attractiveness and extraordinary nutritional and phytochemical composition, is one of the most consumed tropical varieties in the world. A growing body of evidence suggests that their bioactive composition differentiates them from other fruits, with mango pulp being an especially rich and diverse source of polyphenols. In this study, mango pulp polyphenols were submitted to in vitro gastrointestinal digestion and colonic fermentation, and aliquots were analyzed by HPLC-HRMS. The main phenolic compounds identified in the mango pulp were hydroxybenzoic acid-hexoside, two mono-galloyl-glucoside isomers and vanillic acid. The release of total polyphenols increased after the in vitro digestion, with an overall bioaccessibility of 206.3%. Specifically, the most bioaccessible mango polyphenols were gallic acid, 3-O-methylgallic acid, two hydroxybenzoic acid hexosides, methyl gallate, 3,4-dihydroxybenzoic acid and benzoic acid, which potentially cross the small intestine reaching the colon for fermentation by the resident microbiota. After 48 h of fecal fermentation, the main resultant mango catabolites were pyrogallol, gallic and 3,4-dihydroxybenzoic acids. This highlighted the extensive transformation of mango pulp polyphenols through the gastrointestinal tract and by the resident gut microbiota, with the resultant formation of mainly simple phenolics, which can be considered as biomarkers of the colonic metabolism of mango.

Numerous studies have reported health benefits associated with the consumption of fresh and black garlic, which are characterized by the presence of polyphenols and organosulfur compounds (OS). This study aims to analyze the bioaccessibility of the bioactive compounds in fresh and black garlic after in vitro gastrointestinal digestion by monitoring the individual profile of these compounds by ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS). Polyphenols decreased from the beginning of the digestive process, is mainly affected during intestinal digestion. Regarding the OS, the S-alk(en)yl-L-cysteine (SACs) derivatives were more influenced by the acidic conditions of the gastric digestion, while the γ-glutamyl-S-alk(en)yl-L-cysteine (GSAk) derivatives were more susceptible to intestinal digestion conditions in both the fresh and black garlic samples. In conclusion, after in vitro gastrointestinal digestion, the compounds with the highest bioaccessibility were vanillic acid (69%), caffeic acid (52%), γ−glutamyl-S-methyl-L-cysteine sulfoxide (GSMCS) (77%), and S-allylmercapto-L-cysteine (SAMC) (329%) in fresh garlic. Meanwhile, in black garlic, the main bioaccessible compounds were caffeic acid (65%), GSMCS (89%), methionine sulfoxide (262%), trans-S-(1-propenyl)-L-cysteine (151%), and SAMC (106%). The treatment (heating + humidity) to obtain black garlic exerted a positive effect on the bioaccessibility of OS compounds, 55.3% of them remaining available in black garlic, but only 15% in fresh garlic. Polyphenols showed different behavior regarding bioaccessibility.

Background: The t(12;21)(p13;q22), which fuses ETV6 and RUNX1 genes, is the most common genetic abnormality in children with B-cell precursor acute lymphoblastic leukaemia. The implication of the fusion protein in leukemogenesis seems to be clear. However, its role in the maintenance of the disease continues to be controversial. Methods: Generation of an in vitro ETV6/RUNX1 knock out model using the CRISPR/Cas9 gene editing system. Functional characterization by RNA sequencing, proliferation assays, apoptosis and pharmacologic studies, and generation of edited-cell xenograft model. Results: The expression of ETV6/RUNX1 fusion gene was completely eliminated, thus generating a powerful model on which to study the role of the fusion gene in leukemic cells. The loss of fusion gene expression led to the deregulation of biological processes affecting survival such as apoptosis resistance and cell proliferation capacity. Tumour cells showed higher levels of apoptosis, lower proliferation rate and a greater sensitivity to PI3K inhibitors in vitro along as a decrease in tumour growth in xenografts models after ETV6/RUNX1 fusion gene abrogation. Conclusions: ETV6/RUNX1 fusion protein seems to play an important role in the maintenance of the leukemic phenotype and could thus become a potential therapeutic target.

BackgroundEwing Sarcoma (EWS) is a mesenchymal-derived tumor that generally arises in bone and soft tissue. Intensive research regarding the pathogenesis of EWS has been insufficient to pinpoint the early events of Ewing sarcomagenesis. However, the Mesenchymal Stem Cell (MSC) is currently accepted as the most probable cell of origin.Materials and methodsIn an initial study regarding a deep characterization of MSC obtained specifically from EWS patients (MSC-P), we compared them with MSC derived from healthy donors (MSC-HD) and EWS cell lines. We evaluated the presence of the EWS-FLI1 gene fusion and EWSR1 gene rearrangements in MSC-P. The presence of the EWS transcript was confirmed by q-RT-PCR. In order to determine early events possibly involved in malignant transformation, we used a multiparameter quantitative strategy that included both MSC immunophenotypic negative/positive markers, and EWS intrinsic phenotypical features. Markers CD105, CD90, CD34 and CD45 were confirmed in EWS samples.ResultsWe determined that MSC-P lack the most prevalent gene fusion, EWSR1-FLI1 as well as EWSR1 gene rearrangements. Our study also revealed that MSC-P are more alike to MSC-HD than to EWS cells. Nonetheless, we also observed that EWS cells had a few overlapping features with MSC. As a relevant example, also MSC showed CD99 expression, hallmark of EWS diagnosis. However, we observed that, in contrast to EWS cells, MSC were not sensitive to the inhibition of CD99.ConclusionsIn conclusion, our results suggest that MSC from EWS patients behave like MSC-HD and are phenotypically different from EWS cells, thus raising important questions regarding MSC role in sarcomagenesis.

The present study examines the effects of divalent and trivalent Manganese (Mn2+/Mn3+) mixture inhalation on mice to obtain a novel animal model of Parkinson disease (PD) inducing bilateral and progressive dopaminergic cell death, correlate those alterations with motor disturbances, and determine whether L-DOPA treatment improves the behavior, to ensure that the alterations are of dopaminergic origin. CD-1 male mice inhaled a mixture of Manganese chloride and Manganese acetate, one hour twice a week for five months. Before Mn exposure, animals were trained to perform motor function tests and were evaluated each week after the exposure. By the end of Mn exposure, 10 mice were orally treated with 7.5 mg/kg L-DOPA. After 5 months of Mn mixture inhalation, striatal dopamine content decreased 71%, the SNc showed important reduction in the number of TH-immunopositive neurons, mice developed akinesia, postural instability, and action tremor; these motor alterations were reverted with L-DOPA treatment. Our data provide evidence that Mn2+/Mn3+ mixture inhalation produces similar morphological, neurochemical, and behavioral alterations to those observed in PD providing a useful experimental model for the study of this neurodegenerative disease.

Receptor endocytosis is critical for cell signaling. IGF1R mediates an autocrine loop that is de-regulated in Ewing Sarcoma (ES) cells. Here we study the impact of IGF1R internalization, mediated by clathrin and caveolin-1 (CAV1), in ES signaling. We used clathrin and CAV1-siRNA to interfere in clathrin- and caveolin-dependent endocytosis. Chlorpromazine (CPMZ) and methyl-beta-cyclo-dextrin (MCD) were also used in order to inhibit clathrin- and caveolin-dependent endocytosis, respectively. We analyzed IGF1R internalization and co-localization with clathrin and CAV1 upon ligand binding, as well as the status of the IGF1R pathway, cellular proliferation, and the apoptosis of interfered and inhibited ES cells. We performed a high-throughput tyrosine kinase phosphorylation assay to analyze the effects of combining the IGF1R tyrosine kinase inhibitor AEW541 (AEW) with CPMZ or MCD on the intracellular phospho-proteome. We observed that IGF1R is internalized upon ligand binding in ES cells and that this process is dependent on clathrin or CAV1. The blockage of receptor internalization inhibited AKT and MAPK phosphorylation, reducing the proliferative rate of ES cells and increasing the levels of apoptosis. Combination of AEW with CPMZ or MCD largely enhanced these effects. CAV1 and clathrin endocytosis controls IGF1R internalization and signaling and has a profound impact on ES IGF1R-promoted survival signaling. We propose the combination of tyrosine-kinase inhibitors with endocytosis inhibitors as a new therapeutic approach to achieve a stronger degree of receptor inhibition in this, or other neoplasms dependent on IGF1R signaling.