Therapeutic potential of oleic acid supplementation in myotonic dystrophy muscle cell models.

BACKGROUND: We recently reported that upregulation of Musashi 2 (MSI2) protein in the rare neuromuscular disease myotonic dystrophy type 1 contributes to the hyperactivation of the muscle catabolic processes autophagy and UPS through a reduction in miR-7 levels. Because oleic acid (OA) is a known allosteric regulator of MSI2 activity in the biogenesis of miR-7, here we sought to evaluate endogenous levels of this fatty acid and its therapeutic potential in rescuing cell differentiation phenotypes in vitro. In this work, four muscle cell lines derived from DM1 patients were treated with OA for 24 h, and autophagy and muscle differentiation parameters were analyzed. RESULTS: We demonstrate a reduction of OA levels in different cell models of the disease. OA supplementation rescued disease-related phenotypes such as fusion index, myotube diameter, and repressed autophagy. This involved inhibiting MSI2 regulation of direct molecular target miR-7 since OA isoschizomer, elaidic acid (EA) could not cause the same rescues. Reduction of OA levels seems to stem from impaired biogenesis since levels of the enzyme stearoyl-CoA desaturase 1 (SCD1), responsible for converting stearic acid to oleic acid, are decreased in DM1 and correlate with OA amounts. CONCLUSIONS: For the first time in DM1, we describe a fatty acid metabolism impairment that originated, at least in part, from a decrease in SCD1. Because OA allosterically inhibits MSI2 binding to molecular targets, reduced OA levels synergize with the overexpression of MSI2 and contribute to the MSI2 > miR-7 > autophagy axis that we proposed to explain the muscle atrophy phenotype.

Valorization of agri-food waste through the extraction of bioactive molecules. Prediction of their sunscreen action.

The aim of this work was to identify the phenolic composition of 18 different vegetable residues and to determine the relationship between their phenolic compounds, antioxidant capacity and sun protection factor. For this purpose, samples of agri-food residues were analyzed to quantify their antioxidant capacity, total polyphenol and flavonoid content, sun protection factor and individual phenolic compounds through HPLC-DAD-FLD. Among the different phenolic compounds found in the extracts, the phenolic acids, especially caffeic acid, chlorogenic acid, p-coumaric acid and protocatechuic acid were the ones that have been most frequently identified, and, therefore, are present in a wide range of extracts. Black chai tea, lemon ginger tea and peanut extracts were the most antioxidant and photoprotective extracts. Phenolic compounds in the extracts have been found to contribute to their antioxidant activity and are closely correlated to their photoprotective capacity. A regression model that allows predicting the photoprotective capacity of any extract based on its total phenol content has been developed as a tool to determine the most suitable industrial application for each vegetable extract.

Analysis of the Pallial Amygdala in Anurans: Derivatives and Cellular Components.

The amygdaloid complex plays a crucial role in socio-emotional conduct, learning, survival, and reproductive behaviors. It is constituted by a set of nuclei presenting a great cellular heterogeneity and embryonic origin diversity (pallial, subpallial, and even extra-telencephalic). In the last two decades, the tetrapartite pallial paradigm defined the pallial portion of the amygdala as a derivative of the lateroventral pallium. However, the pallial conception is currently being reanalyzed and one of these new proposals is to consider the mouse pallial amygdala as a radial histogenetic domain independent from the rest of the pallial subdomains. In anamniotes, and particularly in amphibian anurans, the amygdaloid complex was described as a region with pallial and subpallial components similar to those described in amniotes. In the present study carried out in Xenopus laevis, after a detailed analysis of the orientation of the amygdalar radial glia, we propose an additional amygdala derived from the pallial region. It is independent of the vomeronasal/olfactory amygdaloid nuclei described in anurans, expresses markers such as Lhx9 present in the mammalian pallial amygdala, and lacks Otp-expressing cells, detected in the adjacent medial amygdala. Further studies are needed to clarify the functional involvement of this area, and whether it is a derivative of the adjacent ventral pallium or an independent pallial domain.

Analysis of the Expression Pattern of Cajal-Retzius Cell Markers in the Xenopus laevis Forebrain.

Cajal-Retzius cells are essential for cortical development in mammals, and their involvement in the evolution of this structure has been widely postulated, but very little is known about their progenitor domains in non-mammalian vertebrates. Using in situhybridization and immunofluorescence techniques we analyzed the expression of some of the main Cajal-Retzius cell markers such as Dbx1, Ebf3, ER81, Lhx1, Lhx5, p73, Reelin, Wnt3a, Zic1, and Zic2 in the forebrain of the anuran Xenopus laevis, because amphibians are the only class of anamniote tetrapods and show a tetrapartite evaginated pallium, but no layered or nuclear organization. Our results suggested that the Cajal-Retzius cell progenitor domains were comparable to those previously described in amniotes. Thus, at dorsomedial telencephalic portions a region comparable to the cortical hem was defined in Xenopus based on the expression of Wnt3a, p73, Reelin, Zic1, and Zic2. In the septum, two different domains were observed: a periventricular dorsal septum, at the limit between the pallium and the subpallium, expressing Reelin, Zic1, and Zic2, and a related septal domain, expressing Ebf3, Zic1, and Zic2. In the lateral telencephalon, the ventral pallium next to the pallio-subpallial boundary, the lack of Dbx1 and the unique expression of Reelin during development defined this territory as the most divergent with respect to mammals. Finally, we also analyzed the expression of these markers at the prethalamic eminence region, suggested as Cajal-Retzius progenitor domain in amniotes, observing there Zic1, Zic2, ER81, and Lhx1 expression. Our data show that in anurans there are different subtypes and progenitor domains of Cajal-Retzius cells, which probably contribute to the cortical regional specification and territory-specific properties. This supports the notion that the basic organization of pallial derivatives in vertebrates follows a comparable fundamental arrangement, even in those that do not have a sophisticated stratified cortical structure like the mammalian cerebral cortex.

Assessment of the determinants of changes and test-retest reliability in the 6-min walk test performance over a 4-month period in healthy 6-12-year-old children.

PURPOSE: The purpose of the study is to establish the determinants of change in 6-min walk test (6MWT) performance observed in children aged 6-12 years over a 4-month period, and to provide test-retest reliability (4 months) to establish the minimal detectable change (MDC). METHODS: Healthy children aged 6-12 years performed two 6MWT trials separated by a period of 4 months. Multiple linear regression analysis was performed to estimate the percentage of variance explained by the variables potentially predictive of the change in the 6MWT. We employed the intraclass correlation coefficient to assess test-retest reliability. RESULTS: Fifty-nine children (28 boys and 31 girls) were assessed. The change in distance covered during the 6MWT was significantly correlated with the growth in their height (r = 0.679; p < 0.05) and the change in their weight (r = 0.473; p < 0.05). Multiple linear regression analysis shows that the change in distance covered in the 6MWT was only explained by its growth in height (46.0% explained variance). The test-retest reliability was fair-good. After 4 months, we established a 12% change from the initial measurement (79.69 m) as the MDC for a 90% confidence level (MDC90). CONCLUSIONS: The distance covered in the 6MWT improved as the children's age, weight and height increased. The growth children's height was the most important predictor of change in distance covered in the 6MWT. An increase of at least 79.69 m (MDC90) in distance covered in the 6MWT is necessary to attribute the improvement to an intervention and not to the individual's growth.

Unraveling the Composition of Insecticidal Crystal Proteins in Bacillus thuringiensis: a Proteomics Approach.

Bacillus thuringiensis (Bt) is the most widely used active ingredient for biological insecticides. The composition of δ-endotoxins (Cry and Cyt proteins) in the parasporal crystal determines the toxicity profile of each Bt strain. However, a reliable method for their identification and quantification has not been available, due to the high sequence identity of the genes that encode the δ-endotoxins and the toxins themselves. Here, we have developed an accurate and reproducible mass spectrometry-based method (liquid chromatography-tandem mass spectrometry-multiple reaction monitoring [LC-MS/MS-MRM]) using isotopically labeled proteotypic peptides for each protein in a particular mixture to determine the relative proportion of each δ-endotoxin within the crystal. To validate the method, artificial mixtures containing Cry1Aa, Cry2Aa, and Cry6Aa were analyzed. Determination of the relative abundance of proteins (in molarity) with our method was in good agreement with the expected values. This method was then applied to the most common commercial Bt-based products, DiPel DF, XenTari GD, VectoBac 12S, and Novodor, in which between three and six δ-endotoxins were identified and quantified in each product. This novel approach is of great value for the characterization of Bt-based products, not only providing information on host range, but also for monitoring industrial crystal production and quality control and product registration for Bt-based insecticides.IMPORTANCEBacillus thuringiensis (Bt)-based biological insecticides are used extensively to control insect pests and vectors of human diseases. Bt-based products provide greater specificity and biosafety than broad-spectrum synthetic insecticides. The biological activity of this bacterium resides in spores and crystals comprising complex mixtures of toxic proteins. We developed and validated a fast, accurate, and reproducible method for quantitative determination of the crystal components of Bt-based products. This method will find clear applications in the improvement of various aspects of the industrial production process of Bt. An important aspect of the production of Bt-based insecticides is its quality control. By specifically quantifying the relative proportion of each of the toxins that make up the crystal, our method represents the most consistent and repeatable evaluation procedure in the quality control of different batches produced in successive fermentations. This method can also contribute to the design of specific culture media and fermentation conditions that optimize Bt crystal composition across a range of Bt strains that target different pestiferous insects. Quantitative information on crystal composition should also prove valuable to phytosanitary product registration authorities that oversee the safety and efficacy of crop protection products.

Fruit and vegetable waste management: Conventional and emerging approaches.

Valorization of Fruit and Vegetable Wastes (FVW) is challenging owing to logistic-related problems, as well as to their perishable nature and heterogeneity, among other factors. In this work, the main existing routes for food waste valorization are critically reviewed. The study focuses on FVW because they constitute an important potential source for valuable natural products and chemicals. It can be concluded that FVW management can be carried out following different processing routes, though nowadays the best solution is to find an adequate balance between conventional waste management methods and some emerging valorization technologies. Presently, both conventional and emerging technologies must be considered in a coordinated manner to enable an integral management of FVW. By doing so, impacts on food safety and on the environment can be minimized whilst wasting of natural resources is avoided. Depending on the characteristics of FVW and on the existing market demand, the most relevant valorization options are extraction of bioactive compounds, production of enzymes and exopolysaccharides, synthesis of bioplastics and biopolymers and production of biofuels. The most efficient emergent processing technologies must be promoted in the long term, in detriment of the conventional ones used nowadays. In consequence, future integral valorization of FVW will probably comprise two stages: direct processing of FVW into value-added products, followed by processing of the residual streams, byproducts and leftover matter by means of conventional waste management technologies.

Therapeutic Applications of Rose Hips from Different Rosa Species.

Rosa species, rose hips, are widespread wild plants that have been traditionally used as medicinal compounds for the treatment of a wide variety of diseases. The therapeutic potential of these plants is based on its antioxidant effects caused by or associated with its phytochemical composition, which includes ascorbic acid, phenolic compounds and healthy fatty acids among others. Over the last few years, medicinal interest in rose hips has increased as a consequence of recent research that has studied its potential application as a treatment for several diseases including skin disorders, hepatotoxicity, renal disturbances, diarrhoea, inflammatory disorders, arthritis, diabetes, hyperlipidaemia, obesity and cancer. In this review, the role of different species of Rosa in the prevention of treatment of various disorders related to oxidative stress, is examined, focusing on new therapeutic approaches from a molecular point of view.

Identification of striatal and pallidal regions in the subpallium of anamniotes.

The telencephalic basal ganglia (BG) of amniotes consist of two subdivisions, striatum and pallidum, which share many features, including development, cell types, neurotransmitter organization and hodology. In particular, these two subdivisions during development are defined on the basis of discrete gene expression patterns (genoarchitecture or genoarchitectonics). The characterization of the BG in the subpallium of representatives of the different classes of anamniote vertebrates was first approached in studies dealing with their localization, hodology and main neurochemical characteristics. Thus, it was proposed that an impressive degree of conservation exists across species. New insights can be gained by the comparative analysis of the expression of conserved transcription factors that distinctly define the striatal and pallidal components of the BG in all vertebrates. In addition, the expression of other genes that characterize neighboring regions of the BG is also useful to define the boundaries of each subdivision. Following this approach, we have analyzed the BG in the brain of juvenile representatives of amphibians, lungfishes, holosteans, Polypteriformes and Chondrichthyes. In addition, we briefly review previous data in teleosts and agnathans. The markers used include islet 1 and Dlx as striatal markers, whereas Nkx2.1 is essential for the identification of the pallidum. In turn, Pax6 and in particular Tbr1 are expressed in the pallium. These markers have been systematically analyzed in combination with neuronal markers of specific subpallial territories and cell populations, such as tyrosine hydroxylase, γ-aminobutyric acid, nitric oxide synthase, substance P and enkephalin. The results highlight that many genes share common distribution patterns and are arranged in conserved combinations whose identification has served to define homologies between components of the BG in distant species.

A detailed 3D MRI brain atlas of the African lungfish Protopterus annectens.

The study of the brain by magnetic resonance imaging (MRI) in evolutionary analyses is still in its incipient stage, however, it is particularly useful as it allows us to analyze detailed anatomical images and compare brains of rare or otherwise inaccessible species, evolutionarily contextualizing possible differences, while at the same time being non-invasive. A good example is the lungfishes, sarcopterygians that are the closest living relatives of tetrapods and thus have an interesting phylogenetic position in the evolutionary conquest of the terrestrial environment. In the present study, we have developed a three-dimensional representation of the brain of the lungfish Protopterus annectens together with a rostrocaudal anatomical atlas. This methodological approach provides a clear delineation of the major brain subdivisions of this model and allows to measure both brain and ventricular volumes. Our results confirm that lungfish show neuroanatomical patterns reminiscent of those of extant basal sarcopterygians, with an evaginated telencephalon, and distinctive characters like a small optic tectum. These and additional characters uncover lungfish as a remarkable model to understand the origins of tetrapod diversity, indicating that their brain may contain significant clues to the characters of the brain of ancestral tetrapods.

Comparative MRI analysis of the forebrain of three sauropsida models.

The study of the brain by magnetic resonance imaging (MRI) allows to obtain detailed anatomical images, useful to describe specific encephalic structures and to analyze possible variabilities. It is widely used in clinical practice and is becoming increasingly used in veterinary medicine, even in exotic animals; however, despite its potential, its use in comparative neuroanatomy studies is still incipient. It is a technology that in recent years has significantly improved anatomical resolution, together with the fact that it is non-invasive and allows for systematic comparative analysis. All this makes it particularly interesting and useful in evolutionary neuroscience studies, since it allows for the analysis and comparison of brains of rare or otherwise inaccessible species. In the present study, we have analyzed the prosencephalon of three representative sauropsid species, the turtle Trachemys scripta (order Testudine), the lizard Pogona vitticeps (order Squamata) and the snake Python regius (order Squamata) by MRI. In addition, we used MRI sections to analyze the total brain volume and ventricular system of these species, employing volumetric and chemometric analyses together. The raw MRI data of the sauropsida models analyzed in the present study are available for viewing and downloading and have allowed us to produce an atlas of the forebrain of each of the species analyzed, with the main brain regions. In addition, our volumetric data showed that the three groups presented clear differences in terms of total and ventricular brain volumes, particularly the turtles, which in all cases presented distinctive characteristics compared to the lizards and snakes.

Expression of SATB1 and SATB2 in the brain of bony fishes: what fish reveal about evolution.

Satb1 and Satb2 belong to a family of homeodomain proteins with highly conserved functional and regulatory mechanisms and posttranslational modifications in evolution. However, although their distribution in the mouse brain has been analyzed, few data exist in other non-mammalian vertebrates. In the present study, we have analyzed in detail the sequence of SATB1 and SATB2 proteins and the immunolocalization of both, in combination with additional neuronal markers of highly conserved populations, in the brain of adult specimens of different bony fish models at key evolutionary points of vertebrate diversification, in particular including representative species of sarcopterygian and actinopterygian fishes. We observed a striking absence of both proteins in the pallial region of actinopterygians, only detected in lungfish, the only sarcopterygian fish. In the subpallium, including the amygdaloid complex, or comparable structures, we identified that the detected expressions of SATB1 and SATB2 have similar topologies in the studied models. In the caudal telencephalon, all models showed significant expression of SATB1 and SATB2 in the preoptic area, including the acroterminal domain of this region, where the cells were also dopaminergic. In the alar hypothalamus, all models showed SATB2 but not SATB1 in the subparaventricular area, whereas in the basal hypothalamus the cladistian species and the lungfish presented a SATB1 immunoreactive population in the tuberal hypothalamus, also labeled with SATB2 in the latter and colocalizing with the gen Orthopedia. In the diencephalon, all models, except the teleost fish, showed SATB1 in the prethalamus, thalamus and pretectum, whereas only lungfish showed also SATB2 in prethalamus and thalamus. At the midbrain level of actinopterygian fish, the optic tectum, the torus semicircularis and the tegmentum harbored populations of SATB1 cells, whereas lungfish housed SATB2 only in the torus and tegmentum. Similarly, the SATB1 expression in the rhombencephalic central gray and reticular formation was a common feature. The presence of SATB1 in the solitary tract nucleus is a peculiar feature only observed in non-teleost actinopterygian fishes. At these levels, none of the detected populations were catecholaminergic or serotonergic. In conclusion, the protein sequence analysis revealed a high degree of conservation of both proteins, especially in the functional domains, whereas the neuroanatomical pattern of SATB1 and SATB2 revealed significant differences between sarcopterygians and actinopterygians, and these divergences may be related to the different functional involvement of both in the acquisition of various neural phenotypes.

Msi2 enhances muscle dysfunction in a myotonic dystrophy type 1 mouse model.

BACKGROUND: Myotonic dystrophy type 1 (DM1) is a rare neuromuscular disease caused by a CTG repeat expansion in the 3' untranslated region of the DM1 protein kinase gene. Characteristic degenerative muscle symptoms include myotonia, atrophy, and weakness. We previously proposed an MSI2>miR-7>autophagy axis whereby MSI2 overexpression repressed miR-7 biogenesis that subsequently de-repressed muscle catabolism through excessive autophagy. Because the DM1 HSALR mouse model expressing expanded CUG repeats shows weak muscle-wasting phenotypes, we hypothesized that MSI2 overexpression was sufficient to promote muscle dysfunction in vivo. METHODS: By means of recombinant AAV murine Msi2 was overexpressed in neonates HSALR mice skeletal muscle to induce DM1-like phenotypes RESULTS: Sustained overexpression of the murine Msi2 protein in HSALR neonates induced autophagic flux and expression of critical autophagy proteins, increased central nuclei and reduced myofibers area, and weakened muscle strength. Importantly, these changes were independent of Mbnl1, Mbnl2, and Celf1 protein levels, which remained unchanged upon Msi2 overexpression. CONCLUSIONS: Globally, molecular, histological, and functional data from these experiments in the HSALR mouse model confirms the pathological role of Msi2 expression levels as an atrophy-associated component that impacts the characteristic muscle dysfunction symptoms in DM1 patients.

Distribution of the transcription factor islet-1 in the central nervous system of nonteleost actinopterygian fish: Relationship with cholinergic and catecholaminergic systems.

Islet-1 (Isl1) is one of the most conserved transcription factors in the evolution of vertebrates, due to its continuing involvement in such important functions as the differentiation of motoneurons, among other essential roles in cell fate in the forebrain. Although its functions are thought to be similar in all vertebrates, the knowledge about the conservation of its expression pattern in the central nervous system goes as far as teleosts, leaving the basal groups of actinopterygian fishes overlooked, despite their important phylogenetic position. In order to assess the extent of its conservation among vertebrates, we studied its expression pattern in the central nervous system of selected nonteleost actinopterygian fishes. By means of immunohistochemical techniques, we analyzed the Isl1 expression in the brain, spinal cord, and sensory ganglia of the cranial nerves of young adult specimens of the cladistian species Polypterus senegalus and Erpetoichthys calabaricus, the chondrostean Acipenser ruthenus, and the holostean Lepisosteus oculatus. We also detected the presence of the transcription factor Orthopedia and the enzymes tyrosine hydroxylase (TH) and choline acetyltransferase (ChAT) to better locate all the immunoreactive structures in the different brain areas and to reveal the possible coexpression with Isl1. Numerous conserved features in the expression pattern of Isl1 were observed in these groups of fishes, such as populations of cells in the subpallial nuclei, preoptic area, subparaventricular and tuberal hypothalamic regions, prethalamus, epiphysis, cranial motor nuclei and sensory ganglia of the cranial nerves, and the ventral horn of the spinal cord. Double labeling of TH and Isl1 was observed in cells of the preoptic area, the subparaventricular and tuberal hypothalamic regions, and the prethalamus, while virtually all motoneurons in the hindbrain and the spinal cord coexpressed ChAT and Isl1. Altogether, these results show the high degree of conservation of the expression pattern of the transcription factor Isl1, not only among fish, but in the subsequent evolution of vertebrates.

Antioxidant Properties of Bioactive Compounds in Fruit and Vegetable Waste.

In recent years, great interest has arisen in the study of compounds with antioxidant activity present in agri-food residues [...].

BlockmiR AONs as Site-Specific Therapeutic MBNL Modulation in Myotonic Dystrophy 2D and 3D Muscle Cells and HSALR Mice.

The symptoms of Myotonic Dystrophy Type 1 (DM1) are multi-systemic and life-threatening. The neuromuscular disorder is rooted in a non-coding CTG microsatellite expansion in the DM1 protein kinase (DMPK) gene that, upon transcription, physically sequesters the Muscleblind-like (MBNL) family of splicing regulator proteins. The high-affinity binding occurring between the proteins and the repetitions disallow MBNL proteins from performing their post-transcriptional splicing regulation leading to downstream molecular effects directly related to disease symptoms such as myotonia and muscle weakness. In this study, we build on previously demonstrated evidence showing that the silencing of miRNA-23b and miRNA-218 can increase MBNL1 protein in DM1 cells and mice. Here, we use blockmiR antisense technology in DM1 muscle cells, 3D mouse-derived muscle tissue, and in vivo mice to block the binding sites of these microRNAs in order to increase MBNL translation into protein without binding to microRNAs. The blockmiRs show therapeutic effects with the rescue of mis-splicing, MBNL subcellular localization, and highly specific transcriptomic expression. The blockmiRs are well tolerated in 3D mouse skeletal tissue inducing no immune response. In vivo, a candidate blockmiR also increases Mbnl1/2 protein and rescues grip strength, splicing, and histological phenotypes.

Development of an irradiation equipment to accelerate the degradation of rosé wine in antique green and flint bottles.

Flint bottles make rosé wines more attractive to the customers and, also, allow them to detect oxidation problems in the color of these wines. Nonetheless, transparent bottles do not protect wines from light. In this work, a device capable of accelerating the degradation of rosé wines using W lamps radiation for short times of exposure has been developed. This equipment has been used to accelerate the color photodegradation of rosé wines, allowing, thus, to identify which parameters can be used as markers of such degradation. The irradiation treatment applied to rosé wines bottled in different types of glass (Flint and Antique Green glass) influenced all the samples. However, the wines treated in Flint bottles displayed more important color variations, especially in color intensity (CI) and hue, than the wines treated in Antique Green bottles. These changes entailed a quality loss of rosé wines that can be appreciated with a naked eye. The yellow component of rosé wines treated in transparent bottles increased the detriment of the red and blue ones. Therefore, color parameters such as CI and a*, together with the total anthocyanin content, seem to be good markers of the loss of quality of rosé wines due to the light effects. The next step will be to find a physical, chemical or physical-chemical protection strategy that, when applied to transparent glass, allow to achieve the light-filtering properties of green glass bottles.

Stability and Bioaccessibility of Phenolic Compounds in Rosehip Extracts during In Vitro Digestion.

Rosehips, particularly dog rose fruits (Rosa canina L.), are a great source of antioxidant compounds, mainly phenolics. However, their health benefits directly depend on the bioaccessibility of these compounds affected by gastrointestinal digestion. Thus, the purpose of this research was to study the impact of gastrointestinal and colonic in vitro digestions on the concentration of total and individual bioaccessible phenolic compounds from a hydroalcoholic extract of rosehips (Rosa canina) and also their antioxidant capacity. A total of 34 phenolic compounds were detected in the extracts using UPLC-MS/MS. Ellagic acid, taxifolin, and catechin were the most abundant compounds in the free fraction, while gallic and p-coumaric acids were the main compounds in the bound phenolic fraction. Gastric digestion negatively affected the content of free phenolic compounds and the antioxidant activity measured using the DPPH radical method. However, there was an enhancement of antioxidant properties in terms of phenolic content and antioxidant activity (DPPH (2,2-diphenyl-1-picrylhydrazyl): 18.01 ± 4.22 mmol Trolox Equivalent (TE)/g; FRAP (Ferric Reducing Antioxidant Power): 7.84 ± 1.83 mmol TE/g) after the intestinal stage. The most bioaccessible phenolic compounds were flavonols (73.3%) and flavan-3-ols (71.4%). However, the bioaccessibility of phenolic acids was 3%, probably indicating that most of the phenolic acids were still bound to other components of the extract. Ellagic acid is an exception since it presented a high bioaccessibility (93%) as it was mainly found in the free fraction of the extract. Total phenolic content decreased after in vitro colonic digestion, probably due to chemical transformations of the phenolic compounds by gut microbiota. These results demonstrated that rosehip extracts have a great potential to be used as a functional ingredient.

Evaluation of grape stems and grape stem extracts for sulfur dioxide replacement during grape wine production.

Sulfur dioxide (SO2), the main preservative in wine, may affect the sensory properties of the wines, as well as cause allergic reactions and headaches in sensitive people. The aim of this work was to evaluate the replacement of SO2 in Tempranillo wines with Mazuelo grape stem products. Five Tempranillo red wines were elaborated: positive control (60 mg/L SO2); negative control with no preservatives; Mazuelo extract (200 mg/L); Mazuelo extract combined with SO2 (100 mg/L + 20 mg/L); and Mazuelo stem (400 mg/L). The oenological parameters, antioxidant capacity, total phenolic (TP), total flavonoids (TF) and total anthocyanins (TA) contents were determined. Additionally, individual phenols were analyzed by HPLC-DAD-FLD. The spectrophotometric analyses showed that the wines had similar antioxidant capacities and concentrations of TP and TF. However, TA was more affected by the lack of SO2 as anthocyanins presented higher concentrations in positive control samples. The concentrations of individual phenols followed a similar path in all samples. Wines containing sulfites were more similar than the other treatments. However, these similarities were not reflected on the sensory analysis performed, as triangle test did not show differences between the wine with extract addition and the positive control wine. Therefore, Mazuelo stem extract could be a possible strategy for SO2 replacement. Nevertheless, further studies are necessary to confirm the potential of grape stem extracts as wine preservative.