Are the BPA analogues an alternative to classical BPA? Comparison between 2D and alternative 3D in vitro neuron model to assess cytotoxic and genotoxic effects.

BPA is used in a wide range of consumer products with very concern toxicological properties. The European Union has restricted its use to protect human health. Industry has substituted BPA by BPA analogues. However, there is a lack of knowledge about their impacts. In this work, BPA and 5 BPA analogues (BPS, BPAP, BPAF, BPFL and BPC) have been studied in classical SH-SY5Y and the alternative 3D in vitro models after 24 and 96 h of exposure. Cell viability, percentage of ROS, cell cycle phases as well as the morphology of the spheroids were measured. The 2D model was more sensitive than the 3D models with differences in cell viability higher than 60% after 24 h of exposure, and different mechanisms of ROS production. After chronic exposure, both models were more affected in comparison to the 24 h exposure. After a recovery time (96 h), the spheroids exposed to 2.5-40 µM were able to recover cell viability and the morphology. Among the BPs tested, BPFL>BPAF>BPAP and >BPC revealed higher toxicological effects, while BPS was the only one with lower effects than BPA. To conclude, the SH-SY5Y 3D model is a suitable candidate to perform more reliable in vitro neurotoxicity tests.

Bioaccessibility and Genoprotective Effect of Melanoidins Obtained from Common and Soft Bread Crusts: Relationship between Melanoidins and Their Bioactivity.

Bread crust constitutes an important by-product of the bakery industry, and its utilization for the isolation of melanoidins to be used as a functional ingredient can enhance its added value and contribute to health. The aim of this study was to evaluate the bioaccessibility, bioactivity, and genoprotective effect of melanoidins derived from bread crust. Bioaccessibility was assessed in gastric, intestinal digestion, and colonic fermentation fractions. The results revealed a relationship between bioaccessible melanoidins and their type (common or soft bread). No cytotoxicity effects were observed for bioaccessible fractions, as assessed by MTT and RTA methods, and they did not affect the distribution of E-cadherin in Caco-2 cells, confirming their ability to maintain membrane integrity. Furthermore, our study demonstrated that the gastrointestinal and colonic fermentation fractions successfully transported across the intestinal barrier, without affecting cell permeability, and showed antioxidant activity on the basolateral side of the cell monolayer. Remarkably, both fractions displayed a significant genoprotective effect in Caco-2 cells. Our findings provide crucial insights into the relationship between the melanoidins and their bioactivity and genoprotective effect. These results demonstrated the potential of bioaccessible melanoidins as valuable bioactive compounds for the development of functional foods, without showing toxic effects on gastrointestinal cells.

Beneficial Effects of Olive Oil Enriched with Lycopene on the Plasma Antioxidant and Anti-Inflammatory Profile of Hypercholesterolemic Patients.

Olive oil and lycopene are foods that have potent antioxidant activity. The objective was to determine the effects of consumption of olive oil enriched with lycopene on oxidative stress biomarkers in hypercholesterolemic subjects. We examined the effects of oil enriched with lycopene extract daily intake during 1 month on plasma antioxidant capacity, lipids profile (triacylgycerols, total cholesterol, cHDL; cLDL, ox-LDL), biomarkers of oxidative stress, and inflammatory markers related with atherosclerosis risk (C-reactive protein (CRP), IL-6; sDC4L) in subjects hypercholesteremics (cholesterol > 220 mg/dL). In the group consuming olive oil-lycopene, significant increases (p < 0.05) in the levels of plasma lycopene concentration (0.146 ± 0.03 versus 0.202 ± 0.04 (µmol/L)), α-carotene (0.166 ± 0.064 versus 0.238 ± 0.07) and in ß-carotene (0.493 ± 0.187 versus 0.713 ± 0.221) were observed. These results are linked with the increases of plasma antioxidants and decreases biomarkers of oxidative stress (carbonyl groups, malondialdehyde and 8-hydroxy-deoxiguanosine) observed in hypercholesterolemic group. In relation to lipid profile, a significant decrease was observed in the levels of ox-LDL (781 ± 302 versus 494 ± 200), remaining unchanged the levels of TG, cholesterol, HDL and LDL-c. Regarding inflammatory biomarkers, the levels of CRP and IL-6 decreased significantly. The positive results obtained in this study support the use of olive oil enriched with lycopene to reduce the risk of coronary disease.

Biomarkers of Oxidative Stress in Healthy Infants within the First Three Days after Birth.

The clinical relevance of stress biomarkers in newborns is well established. Currently, oxidative stress (OS) parameters are seen to play an important role in neonatal resuscitation guidelines, and a link has been observed between the amount of oxygen delivered and the level of OS and the development of various pathologies. The aim of the current study was to investigate changes in neonatal plasma and urine OS status during the first hours after birth. A lower antioxidant capacity (TAC) and higher levels of malondialdehyde in blood were observed in newborns at the time of birth compared with results 48 h postnatally. The urine revealed a significant and progressive increase in TAC and creatinine during the first 36 h of life, with a progressive decline thereafter. Meanwhile, malondialdehyde in urine samples showed no significant differences over time. Overall, the correlation between blood and urine parameters was poor, except for the relationship between umbilical vein glutathione reduced/oxidized ratio and urine malondialdehyde (r = 0.7; p = 0.004) and between TAC in the umbilical artery and urine (r = -0.547; p = 0.013). The biomarkers evaluated in this study could be established as reference values for neonatal OS.

Influence of the packaging systems on the phenolic profile and antioxidant properties of wine pomace used as seasoning in chicken meat.

White wine pomace products (wWPP) represent an innovative strategy as a functional food ingredient to be used as a seasoning both for their technological and functional properties. Nevertheless, the bioactive compounds of wWPP used as a seasoning could be modified during storage. The seasoning in the meat, regardless of the storage method used, modified its phenolic profile and in its bioaccessible fractions, while maintaining a high total antioxidant capacity and total polyphenol content. The contact of the seasoning with the meat can be considered safe as it does not show cytotoxicity in the Caco-2 cells. Additionally, the ability to modulate the cell oxidative stress of the bioaccessible fractions and the potential benefits on microbiota by the colonic fermentation fraction, suggest its potential use as a functional ingredient, without being affected by storage. These results are novel and may help to establish the value of this product as a functional ingredient.

Bread melanoidins as potential new sustainable bakery ingredients: a study using fat and fat-free bakery food models.

Melanoidins isolated from bakery by-products are proposed as new sustainable ingredients for bakery products. The colour, odour profile, texture, water activity, and antioxidant capacity of two bakery food models, fat and fat-free, enriched with 2% and 4% soft bread and common bread melanoidins, were analysed. The colour of the bakery food models with melanoidins was darker than that of the respective control; the fat-free models with melanoidins showed higher values of hardness than the control, while no significant effect was observed in the fat models; the water activity did not change compared to the control; the odour profile was significantly modified with different effects depending on the type of melanoidin quantity added and the food model (fat or fat-free); and the antioxidant capacity increased proportionally to the quantity of melanoidin added. In general, melanoidins from soft bread exhibited a higher effect than the melanoidins from common bread. The melanoidins isolated from both fat and fat-free bakery food models did not show cytotoxicity nor did they modify the levels of reactive oxygen species in Caco-2 cells. Therefore, the results seem to indicate the favourable potential of bread melanoidins as new sustainable ingredients for bakery products.

Multi-omic alterations of the SWI/SNF complex define a clinical subgroup in lung adenocarcinoma.

SWI/SNF complexes are major targets of mutations in cancer. Here, we combined multiple "-omics" methods to assess SWI/SNF composition and aberrations in LUAD. Mutations in lung SWI/SNF subunits were highly recurrent in our LUAD cohort (41.4%), and over 70% of the mutations were predicted to have functional impact. Furthermore, SWI/SNF expression in LUAD suffered an overall repression that could not be explained exclusively by genetic alterations. Finally, SWI/SNF mutations were associated with poorer overall survival in TCGA-LUAD. We propose SWI/SNF-mutant LUAD as a separate clinical subgroup with practical implications.

From winery by-product to healthy product: bioavailability, redox signaling and oxidative stress modulation by wine pomace product.

The wine pomace is the main winery by-products that suppose an economic and environmental problem and their use as a functional ingredient are being increasingly recognized as a good and inexpensive source of bioactive compounds. In this sense, it is known the potential health properties of wine pomace products in the prevention of disorders associated with oxidative stress and inflammation such as endothelial dysfunction, hypertension, hyperglycemia, diabetes, obesity. Those effects are due to the bioactive compounds of wine pomace and the mechanisms concern especially modulation of antioxidant/prooxidant activity, improvement of nitric oxide bioavailability, reduction of pro-inflammatory cytokines and modulation of antioxidant/inflammatory signal pathways. This review mainly summarizes the mechanisms of wine pomace products as modulators of oxidative status involved in cell pathologies as well as their potential therapeutic use for cardiovascular diseases. For this purpose, the review provides an overview of the findings related to the wine pomace bioactive compounds profile, their bioavailability and the action mechanisms for maintaining the redox cell balance involved in health benefits. The review suggests an important role for wine pomace product in cardiovascular diseases prevention and their regular food intake may attenuate the development and progression of comorbidities associated with cardiovascular diseases.

Effects of Hypothermia and Allopurinol on Oxidative Status in a Rat Model of Hypoxic Ischemic Encephalopathy.

Hypoxic ischemic encephalopathy (HIE) is one of the main causes of morbidity and mortality during the neonatal period, despite treatment with hypothermia. There is evidence that oxidative damage plays an important role in the pathophysiology of hypoxic-ischemic (HI) brain injury. Our aim was to investigate whether postnatal allopurinol administration in combination with hypothermia would reduce oxidative stress (OS) biomarkers in an animal model of HIE. Postnatal 10-day rat pups underwent unilateral HI of moderate severity. Pups were randomized into: Sham operated, hypoxic-ischemic (HI), HI + allopurinol (HIA), HI + hypothermia (HIH), and HI + hypothermia + allopurinol (HIHA). Biomarkers of OS and antioxidants were evaluated: GSH/GSSG ratio and carbonyl groups were tested in plasma. Total antioxidant capacity (TAC) was analyzed in plasma and cerebrospinal fluid, and 8-iso-prostaglandin F2α was measured in brain tissue. Plasma 2,2'-azinobis-(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) levels were preserved in those groups that received allopurinol and dual therapy. In cerebrospinal fluid, only the HIA group presented normal ferric reducing ability of plasma (FRAP) levels. Protein oxidation and lipid peroxidation were significantly reduced in all groups treated with hypothermia and allopurinol, thus enhancing neuroprotection in HIE.

Wine Pomace Product Inhibit Listeria monocytogenes Invasion of Intestinal Cell Lines Caco-2 and SW-480.

Red wine pomace products (WPP) have antimicrobial activities against human pathogens, and it was suggested that they have a probable anti-Listeria effect. This manuscript evaluates the intestinal cell monolayer invasive capacity of Listeria monocytogenes strains obtained from human, salmon, cheese, and L. innocua treated with two WPP (WPP-N and WPP-C) of different polyphenol contents using Caco-2 and SW480 cells. The invasion was dependent of the cell line, being higher in the SW480 than in the Caco-2 cell line. Human and salmon L. monocytogenes strains caused cell invasion in both cell lines, while cheese and L. innocua did not cause an invasion. The phenolic contents of WPP-N are characterized by high levels of anthocyanin and stilbenes and WPP-C by a high content of phenolic acids. The inhibitory effect of the WPPs was dependent of the strain and of the degree of differentiation of the intestinal cells line. The inhibition of Listeria invasion by WPPs in the SW480 cell line, especially with WPP-C, were higher than the Caco-2 cell line inhibited mainly by WPP-N. This effect is associated with the WPPs' ability to protect the integrity of the intestinal barrier by modification of the cell-cell junction protein expression. The gene expression of E-cadherin and occludin are involved in the L. monocytogenes invasion of both the Caco-2 and SW480 cell lines, while the gene expression of claudin is only involved in the invasion of SW480. These findings suggest that WPPs have an inhibitory L. monocytogenes invasion effect in gastrointestinal cells lines.

Phyllosilicate-content influence on the spectroscopic properties and antioxidant capacity of Iberian Cretaceous clays.

Kaolinite-rich Cretaceous clay sediment samples from Burgos (Spain) have been analyzed by elemental analysis, X-ray fluorescence, inductively coupled plasma mass spectrometry, X-ray diffraction and different spectroscopic techniques, as Fourier Transform Infrared, ultraviolet-visible and electron paramagnetic resonance. The clay sediment samples mainly contain quartz, muscovite and kaolinite. Different radicals, as A- and B-Centers in kaolinite and organic paramagnetic species, are detected. An illite/kaolinite FTIR band ratio parameter (IKB) is proposed to infer the illite/kaolinite proportion, which can be useful to graphically visualize the iron-substituted Al(III) sites. Studies of the activity as scavengers of DPPH and ABTS radicals show that samples with a larger amount of orthorhombic Fe(III) ions replacing Al(III) ions exhibit a higher antioxidant capacity.

Cytotoxicity study of bakery product melanoidins on intestinal and endothelial cell lines.

Melanoidins contribute to organoleptic properties of processed foods and exert benefits in health. The aim of this study was to isolate and characterize melanoidins from baked products (common bread, soft bread and biscuits), evaluate their cytotoxicity and determine their suitability as functional additives. Extraction yield, spectrophotometric characteristics, colorimetric properties, antioxidant capacity, and cytotoxicity of melanoidins were assessed. Among the studied products, soft bread had the highest extraction throughput. Melanoidins from biscuit showed the highest antioxidant capacity, closely followed by those of soft bread. Melanoidins did not exert cytotoxic effects on Caco-2 and HUVEC cells (viability was >80%). Nevertheless, incubation of HUVEC cells with melanoidins from common bread and biscuit slightly decreased viability, whereas gastrointestinal digestion of such melanoidins softened the decrease in cell viability. This study point to soft bread as a safe and efficient source of melanoidins, that could be potentially used in the future as functional ingredient.

Bioavailable wine pomace attenuates oxalate-induced type II epithelial mesenchymal transition and preserve the differentiated phenotype of renal MDCK cells.

The functional renal epithelium is composed of differentiated and polarized tubular cells with a strong actin cortex and specialized cell-cell junctions. If, under pathological conditions, these cells have to resist higher kidney osmolarity, they need to activate diverse mechanisms to survive external nephrotoxic agents such as inflammation and oxidative stress. Wine pomace polyphenols exert protective effects on renal cells. In this study, two wine-pomace products and their protective effects upon promotion and preservation of normal cell differentiation and attenuation of oxalate-induced type II epithelial mesenchymal transition (EMT) are evaluated. Treatment with gastrointestinal and colonic bioavailable fractions from red (rWPP) and white (wWPP) wine pomaces, both in the presence and the absence of oxalate, showed similar cell numbers and nuclear size than the non-treated differentiated MDCK cells. Immunofluorescence analysis showed the reduction of morphological changes and the preservation of cellular junctions for the rWPP and wWPP pre-treatment of cells exposed to oxalate injury. Hence, both rWPP and wWPP attenuated oxalate type II EMT in MDCK cells that conserved their epithelial morphology and cellular junctions through the antioxidant activities of grape pomace polyphenols.

The protective effects of wine pomace products on the vascular endothelial barrier function.

Endothelial dysfunction is associated with cardiovascular diseases and involves a chronic inflammatory process that together with oxidative stress increases the permeability of the vascular endothelium. The aim of this study was to evaluate the role of red and white wine pomace products (rWPPs and wWPPs) in the maintenance of endothelial integrity in hyperglycemia of EA.hy926 endothelial cells. EA.hy926 endothelial cells exposed to hyperglycemia were treated with the in vitro digested fractions of rWPPs and wWPPs. A Real Time Cellular Analysis (RTCA) system was used to evaluate the endothelial monolayer integrity after INF-γ stimulation of pre-treated endothelial cells with the digested fractions. The changes in cell viability, NO, ROS and NOX4 were recorded and actin cytoskeleton and E-cadherin junctions were evaluated by immunofluorescence. All digested fractions prevent the hyperglycemic actions in the cell viability and NO/ROS balance. The inflammatory mediator INF-γ and hyperglycemia caused a decrease in RTCA adhesion of the EA.hy926 endothelial cell monolayer. Pre-treatment with all digested fractions enhanced the EA.hy926 endothelial monolayer integrity and maintained actin cytoskeleton and E-cadherin junctions. These in vitro studies elucidate that the anti-hyperglycemic and anti-inflammatory actions of wine pomace products involve a decrease in ROS production and the stabilization of junction proteins via modulation of VE-cadherin and actin cytoskeleton suggesting a potential prevention of endothelial damage by these natural products.

The value of lncRNA FENDRR and FOXF1 as a prognostic factor for survival of lung adenocarcinoma.

It is increasingly evident that non-coding RNAs play a significant role in tumour development. However, we still have a limited knowledge of the clinical significance of long non-coding RNAs (lncRNAs) in lung cancer. The FENDRR is a long coding RNA (also named FOXF1-AS1) located in the vicinity of the protein-coding gene FOXF1 at 16q24.1 chromosomal region. The present study aimed to define the clinic pathological significance of the long-non-coding RNA FENDRR in lung adenocarcinomas. FENDRR expression measured by quantitative PCR was found significantly downregulated (p<0.001) in lung adenocarcinoma samples in comparison with their normal adjacent tissues (n=70). RNA in situ hybridization (RNA-FISH) corroborated independently the down-regulation of FENDRR. Interestingly, the expression of FENDRR correlated positively (p<0.001) with the expression of its protein-coding neighbor gene FOXF1. Additionally, FOXF1 expression was also found downregulated in adenocarcinomas compared to normal samples (p<0.001) and its expression was significantly correlated with overall survival alone (p=0.003) or in combination with FENDRR expression (p=0.01). In conclusion, our data support that FENDRR and FOXF1 expression is decreased in lung adenocarcinoma and should be considered as new potential diagnostic/prognosis biomarkers.

The dose-response effect on polyphenol bioavailability after intake of white and red wine pomace products by Wistar rats.

Wine pomace by-products are an important source of phenolic acids with significant health benefits. However, phenolic acid bioavailability in vivo has been little studied and there are few comparative studies on bioavailability between red and white wine pomace and the effect of intake of different doses. The qualitative and quantitative profile of phenolic acid metabolites in plasma and urine samples from Wistar rats was obtained by gas chromatography/mass detection, after oral administration of four doses (50, 100, 150, and 300 mg) of both the red and the white wine pomace products (rWPP and wWPP, respectively). The antioxidant capacity of the plasma samples assessed by both the ABTS and the FRAP levels was also evaluated. The results showed that neither the bioavailability nor the antioxidant capacity in vivo of the rWPP increased at high doses. However, both parameters were dependent on the intake of the wWPP.

Assessment of a New ROS1 Immunohistochemistry Clone (SP384) for the Identification of ROS1 Rearrangements in Patients with Non-Small Cell Lung Carcinoma: the ROSING Study.

INTRODUCTION: The ROS1 gene rearrangement has become an important biomarker in NSCLC. The College of American Pathologists/International Association for the Study of Lung Cancer/Association for Molecular Pathology testing guidelines support the use of ROS1 immunohistochemistry (IHC) as a screening test, followed by confirmation with fluorescence in situ hybridization (FISH) or a molecular test in all positive results. We have evaluated a novel anti-ROS1 IHC antibody (SP384) in a large multicenter series to obtain real-world data. METHODS: A total of 43 ROS1 FISH-positive and 193 ROS1 FISH-negative NSCLC samples were studied. All specimens were screened by using two antibodies (clone D4D6 from Cell Signaling Technology and clone SP384 from Ventana Medical Systems), and the different interpretation criteria were compared with break-apart FISH (Vysis). FISH-positive samples were also analyzed with next-generation sequencing (Oncomine Dx Target Test Panel, Thermo Fisher Scientific). RESULTS: An H-score of 150 or higher or the presence of at least 70% of tumor cells with an intensity of staining of 2+ or higher by the SP384 clone was the optimal cutoff value (both with 93% sensitivity and 100% specificity). The D4D6 clone showed similar results, with an H-score of at least 100 (91% sensitivity and 100% specificity). ROS1 expression in normal lung was more frequent with use of the SP384 clone (p < 0.0001). The ezrin gene (EZR)-ROS1 variant was associated with membranous staining and an isolated green signal FISH pattern (p = 0.001 and p = 0.017, respectively). CONCLUSIONS: The new SP384 ROS1 IHC clone showed excellent sensitivity without compromising specificity, so it is another excellent analytical option for the proposed testing algorithm.

Revisiting the thiosemicarbazonecopper(II) reaction with glutathione. Activity against colorectal carcinoma cell lines.

Thiosemicarbazones (TSCs), and their copper derivatives, have been extensively studied mainly due to the potential applications as antitumor compounds. A part of the biological activity of the TSC-CuII complexes rests on their reactivity against cell reductants, as glutathione (GSH). The present paper describes the structure of the [Cu(PTSC)(ONO2)]n compound (1) (HPTSC=pyridine-2-carbaldehyde thiosemicarbazone) and its spectroscopic and magnetic properties. ESI studies performed on the reaction of GSH with 1 and the analogous [{Cu(PTSC*)(ONO2)}2] derivative (2, HPTSC*=pyridine-2-carbaldehyde 4N-methylthiosemicarbazone) show the absence of peaks related with TSC-Cu-GSH species. However GSH-Cu ones are detected, in good agreement with the release of CuI ions after reduction in the experimental conditions. The reactivity of 1 and 2 with cytochrome c and myoglobin and their activities against HT-29 and SW-480 colon carcinoma cell lines are compared with those shown by the free HPTSC and HPTSC* ligands.

Irrigation water consumption modelling of a soilless cucumber crop under specific greenhouse conditions in a humid tropical climate / Modelação de consumo de água de irrigação de pepino sem solo em condições específicas de uma casa de vegetação em um clima tropical húmido

ABSTRACT: The irrigation water consumption of a soilless cucumber crop under greenhouse conditions in a humid tropical climate has been evaluated in this paper in order to improve the irrigation water and fertilizers management in these specific conditions. For this purpose, a field experiment was conducted. Two trials were carried out during the years 2011 and 2014 in an experimental farm located in Vinces (Ecuador). In each trial, the complete growing cycle of a cucumber crop grown under a greenhouse was evaluated. Crop development was monitored and a good fit to a sigmoidal Gompertz type growth function was reported. The daily water uptake of the crop was measured and related to the most relevant indoor climate variables. Two different combination methods, namely the Penman-Monteith equation and the Baille equation, were applied. However, the results obtained with these combination methods were not satisfactory due to the poor correlation between the climatic variables, especially the incoming radiation, and the crop's water uptake (WU). On contrary, a good correlation was reported between the crop's water uptake and the leaf area index (LAI), especially in the initial crop stages. However, when the crop is fully developed, the WU stabilizes and becomes independent from the LAI. A preliminary model to simulate the water uptake of the crop was adjusted using the data obtained in the first experiment and then validated with the data of the second experiment.

RESUMO: Com o objetivo de melhorar a gestão da água de irrigação e fertilizantes em condições específicas, foi avaliado, neste trabalho, o consumo de água de irrigação de pepinos cultivados sem solo em casa de vegetação. Para avaliar este efeito, conduziu-se um experimento de campo. O trabalho de pesquisa foi composto de dois ensaios realizados durante os anos de 2011 e 2014 em Vinces (Equador), onde predomina o clima tropical. Em cada ensaio, avaliou-se o ciclo de crescimento completo da cultura do pepino cultivada em estufa.O desenvolvimento da cultura foi monitorado e um bom ajuste para a função de crescimento tipo sigmoidal Gompertz foi encontrado. A absorção de água diária da cultura foi medida e relacionada às variáveis climáticas mais relevantes. Com o objetivo de modelar o consumo de água das culturas, dois métodos de combinação diferentes, isto é, a equação de Penman-Monteith e a equação Baille, foram aplicadas. No entanto, os resultados obtidos com estes métodos de combinação não foram satisfatórios devido a fraca correlação entre as variáveis climáticas, especialmente, a radiação incidente e absorção de água pela cultura (WU). Por outro lado, foi encontrada uma boa correlação entre a captação de água da cultura e o índice de área foliar (LAI), especialmente, nos estágios iniciais da cultura. No entanto, quando a cultura está completamente desenvolvida, o WU estabiliza e torna-se independente do LAI. Um modelo preliminar para simular a absorção de água da cultura foi ajustado e utilizado e, os dados obtidos do primeiro experimento foram comparados com os dados do segundo.