Supercritical CO2 Processing of White Grape Must as a Strategy to Reduce the Addition of SO2.

In winemaking, sulfur dioxide addition is the most common procedure to prevent enzymatic and microbial alterations. However, the enological industry looks for safer alternatives to preserve enological products, and high-pressure treatments with supercritical CO2 are a suitable alternative. This study evaluates the effectiveness of this process in the stabilization and preservation of white grape must, studying the influence of time, pressure, and CO2 percentage on must characteristics. In spite of the percentage of CO2 turned out to be the variable that affects the most the process, no remarkable differences were observed in pH, acidity, and color intensity between untreated and treated musts. Moreover, this technique has proven to be very efficient in the reduction of aerobic mesophilic microorganisms as well as in the reduction of residual polyphenol oxidase activities, being lower than those obtained with SO2 addition (60 and 160 mg/L). Based on the results, the most convenient conditions were 100 bar and 10% CO2, for 10 min treatment.

Functional, Physical, and Volatile Characterization of Chitosan/Starch Food Films Functionalized with Mango Leaf Extract.

Active packaging is one of the currently thriving methods to preserve highly perishable foods. Nonetheless, the integration of active substances into the formulation of the packaging may alter their properties-particularly mass transfer properties-and therefore, the active compounds acting. Different formulations of chitosan (CH), starch (ST), and their blends (CH-ST), with the addition of mango leaf extract (MLE) have been polymerized by casting to evaluate their food preservation efficiency. A CH-ST blend with 3% MLE using 7.5 mL of the filmogenic solution proved to be the most effective formulation because of its high bioactivity (ca. 80% and 74% of inhibition growth of S. aureus and E. coli, respectively, and 40% antioxidant capacity). The formulation reduced the water solubility and water vapor permeability while increasing UV protection, properties that provide a better preservation of raspberry fruit after 13 days than the control. Moreover, a novel method of Headspace-Gas Chromatography-Ion Mobility Spectrometry to analyze the volatile profiles of the films is employed, to study the potential modification of the food in contact with the active film. These migrated compounds were shown to be closely related to both the mango extract additions and the film's formulation themselves, showing different fingerprints depending on the film.

Use of Winemaking By-Products for the Functionalization of Polylactic Acid for Biomedical Applications.

The addition of naturally active compounds to implantable polymers is an efficient strategy against inflammation issues that might lead to rejection, while promoting controlled re-endothelialization of the tissues. This work proposes the use of winemaking by-products with high active properties of biomedical interest to obtain bioactive PLA by using supercritical technologies. First, two red grape pomace extracts, obtained by high-pressure extraction with supercritical CO2 and cosolvents (either ethanol or water-ethanol), have been studied. Second, two impregnation methods have been studied with both extracts, traditional supercritical CO2-assisted impregnation (SSI) and a novel pressurized soaking method (PSI). The amount of extract impregnated as well as the bioactivity levels achieved-i.e., antioxidant, antimicrobial, and anti-inflammatory properties- have been determined for each extract and impregnation method at different pressure and temperature conditions. Both extracts obtained had good antioxidant, anti-inflammatory, and antibacterial capacities, especially the hydroethanolic one (0.50 ± 0.03 mg TE/g versus 0.24 ± 0.03 mg TE/g, respectively). Regarding impregnated filaments, impregnation loadings depended especially on the extract and P/T conditions, providing up to 8% (extract mass/polymer mass) of impregnation. The antioxidant capacity increased noteworthily by using the ethanolic extract by PSI, with values near 100 µg TE/g PLA.

The effects of mango leaf extract during adolescence and adulthood in a rat model of schizophrenia.

There is evidence that in schizophrenia, imbalances in inflammatory and oxidative processes occur during pregnancy and in the early postnatal period, generating interest in the potential therapeutic efficacy of anti-inflammatory and antioxidant compounds. Mangiferin is a polyphenolic compound abundant in the leaves of Mangifera indica L. that has robust antioxidant and anti-inflammatory properties, making it a potential candidate for preventive or co-adjuvant therapy in schizophrenia. Hence, this study set-out to evaluate the effect of mango leaf extract (MLE) in a model of schizophrenia based on maternal immune activation, in which Poly I:C (4 mg/kg) is administered intravenously to pregnant rats. Young adult (postnatal day 60-70) or adolescent (postnatal day 35-49) male offspring received MLE (50 mg/kg of mangiferin) daily, and the effects of MLE in adolescence were compared to those of risperidone, assessing behavior, brain magnetic resonance imaging (MRI), and oxidative/inflammatory and antioxidant mediators in the adult offspring. MLE treatment in adulthood reversed the deficit in prepulse inhibition (PPI) but it failed to attenuate the sensitivity to amphetamine and the deficit in novel object recognition (NOR) induced. By contrast, adolescent MLE treatment prevented the sensorimotor gating deficit in the PPI test, producing an effect similar to that of risperidone. This MLE treatment also produced a reduction in grooming behavior, but it had no effect on anxiety or novel object recognition memory. MRI studies revealed that adolescent MLE administration partially counteracted the cortical shrinkage, and cerebellum and ventricle enlargement. In addition, MLE administration in adolescence reduced iNOS mediated inflammatory activation and it promoted the expression of biomarkers of compensatory antioxidant activity in the prefrontal cortex and hippocampus, as witnessed through the reduction of Keap1 and the accumulation of NRF2 and HO1. Together, these findings suggest that MLE might be an alternative therapeutic or preventive add-on strategy to improve the clinical expression of schizophrenia in adulthood, while also modifying the time course of this disease at earlier stages in populations at high-risk.

Supercritical Impregnation of Mango Leaf Extract into PLA 3D-Printed Devices and Evaluation of Their Biocompatibility with Endothelial Cell Cultures.

The addition of natural substances with pharmacoactive properties to polymeric biomedical devices would provide beneficial regarding the assimilation of these endoprostheses when implanted into a patient's body. The added drug would facilitate endothelization by regulating the inflammatory processes that such interventions entail, preventing contamination hazards and favoring the angiogenesis or formation of blood vessels in the tissue. The present work used mango leaf extract (MLE) obtained through pressurized ethanol for this purpose. Polylactic acid (PLA) in the form of filaments or 3D-printed disks was impregnated by means of supercritical technology with MLE for the culture essays. The release kinetics has been studied and the polymer matrices have been examined by scanning electron microscopy (SEM). The impregnated devices were subjected to in vitro culture of colony-forming endothelial cells. The influence of the different impregnation conditions used for the production of the MLE impregnated polymeric devices on the development of the cell culture was determined by fluorescence microscopy. The best results were obtained from the calcein cultures on 35 °C MLE impregnated into 3D-printed polymer disks.

Structural Modification of Polymers Functionalized with Mango Leaf Extract by Supercritical Impregnation: Approaching of Further Food and Biomedical Applications.

Identifying new polymers from natural resources that can be effectively functionalized can have a substantial impact on biomedical devices and food preservation fields. Some of these polymers would be made of biodegradable, renewable and compostable materials, and present the kind of porosity required to effectively carry active compounds that confer on them the desired properties for their intended applications. Some natural extracts, such as mango leaf extract, have been proven to have high levels of antioxidant, antimicrobial or anti-inflammatory properties, making them good candidates for controlled-release applications. This work intends to investigate the supercritical impregnation of different types of polymers (ABS, PETG, TPU, PC and PCL) with mango leaf extract. The influence of temperature and pressure on the polymers' structure (swelling and foaming processes) and their different behaviors have been analyzed. Thus, TPU and PC experience minimal structural modifications, while PETG, PCL and ABS, on the other hand, suffer quite significant structural changes. TPU and PETG were selected as the representative polymers for each one of these behaviors to delve into mango leaf extract impregnation processes. The bioactive capacity of the extract is present in either impregnated polymer, with 25.7% antioxidant activity by TPU processed at 35 °C and 100 bar and 32.9% antioxidant activity by PETG impregnated at 75 °C and 400 bar.

Pro-Angiogenic Effects of Natural Antioxidants Extracted from Mango Leaf, Olive Leaf and Red Grape Pomace over Endothelial Colony-Forming Cells.

Cardiovascular diseases remain the leading cause of death worldwide, mainly triggered by the formation of atherosclerotic plaques that reduce blood flow. Angiogenic cell therapy based on endothelial colony forming cells (ECFCs) constitutes a promising alternative to promote vascular revascularization; however, under the oxidative environment that prevails in ischemic areas, these cells become impaired. Thus, it is necessary to investigate strategies to enhance their regenerative properties. Antioxidant substances, such as polyphenols, have been shown to be useful for this purpose. In the current study we evaluated the potential of mango leaves, olive leaves and red grape pomace extracts, rich in polyphenols, to promote ECFC reparative effects. For this, aqueous and ethanolic extracts of the aforementioned raw materials were obtained by pressurized liquid extraction (PLE). After evaluating the polyphenol content and the antioxidant activity, in vitro assays were carried out, and we found that ethanolic extracts at low concentrations improved angiogenic capacities of ECFCs and reduced proliferation, apoptosis, and the inflammatory response of these cells. Overall, mango leaves ethanolic extract provided the most promising results, but all three extracts ameliorated the functionality of ECFCs.

Health-Promoting Properties of Borage Seed Oil Fractionated by Supercritical Carbon Dioxide Extraction.

Borage (Borago officinalis L.) seed oil is an important source of γ-linolenic acid, which is normally used as a treatment against different pathologies. Since the fractionation of this interesting seed oil has many environmental, economic and biological benefits, two borage fractionation techniques after extraction with CO2 under supercritical conditions have been studied: precipitation in two cyclone separators and countercurrent extraction column. Both techniques have successfully collected free fatty acids in one fraction: (i) two separators set up in series obtained the highest concentration of free fatty acids in separator 2 at 90 bar/40 °C; (ii) when countercurrent extraction column was used, the acidity index of the raffinate stream was independent from the operating conditions (2.6 ± 0.5%). Furthermore, the composition of the fatty acids, as well as their antioxidant and cytotoxic activities, were determined. The profile of the fatty acids obtained by either of these two methods remained unaltered, so that the crude oil exhibited improved antioxidant and cytotoxic properties. All the extracts obtained in the two cyclone separators at the same pressure/temperature conditions displayed high tumouricidal activity against HL 60 promyelocytic leukaemia cells, even if the extracts at 50% concentration from separator 2 presented a lower inhibitory activity (IC50). The extracts from separator 2 at 90 bar/40 °C exhibited the highest anti-proliferative activity at low doses (IC50 of 0.3 µL/mL for the trypan blue exclusion test). To reach the lethal dose-IC50-with the product obtained through countercurrent column fractionation, a concentration of 2 µL/mL of crude borage oil raffinate was required.

Potential Use of Annona Genus Plants Leaf Extracts to Produce Bioactive Transdermal Patches by Supercritical Solvent Impregnation.

The objective of the present work was to develop a bioactive transdermal patch functionalized with Annona leaf extracts (ALE) by means of supercritical impregnation technique. The potential of six different Annona leaf extracts (ALE) obtained with the enhanced solvent system formed by carbon dioxide + ethanol/acetone was evaluated taking into account the antioxidant activity, total phenol composition and global extraction yields. For the impregnation of ALE, two drug supporting systems were tested: hydrocolloid sodium carboxymethyl cellulose (NaCMC) and polyester dressings (PD). The effect of the impregnation conditions, including pressure (P), temperature (T), percent of co-solvent (ethanol) and ALE/polymer mass ratio, was determined with regard to the loading and the functional activity of the impregnated samples. The optimal impregnation conditions of ALE were established at 55 °C and 300 bar which led to obtained transdermal patches with antioxidant and antimicrobial capacity. In order to understand the behavior of the process, the homogeneity of the samples in the vessels was also evaluated. The best results were obtained at higher proportions of co-solvent in the system.

Supercritical Impregnation of PLA Filaments with Mango Leaf Extract to Manufacture Functionalized Biomedical Devices by 3D Printing.

Polylactic Acid (PLA) filaments impregnated with ethanolic mango leaves extract (MLE) with pharmacological properties were obtained by supercritical impregnation. The effects of pressure, temperature and amount of extract on the response variables, i.e., swelling, extract loading and bioactivity of the PLA filaments, were determined. The analysis of the filaments biocapacities revealed that impregnated PLA filaments showed 11.07% antidenaturant capacity and 88.13% antioxidant activity, which after a 9-day incubation shifted to 30.10% and 9.90%, respectively. Subsequently, the same tests were conducted on printed samples. Before their incubation, the printed samples showed 79.09% antioxidant activity and no antidenaturant capacity was detected. However, after their incubation, the antioxidant activity went down to only 2.50%, while the antidenaturant capacity raised up to 23.50%. The persistence of the bioactive properties after printing opens the possibility of using the functionalized PLA filaments as the feed for a three-dimensional (3D) printer.

Application of a Natural Antioxidant from Grape Pomace Extract in the Development of Bioactive Jute Fibers for Food Packaging.

There is an increasing demand for the use of new food packaging materials. In this study, natural jute fibers impregnated with a Petit Verdot Red Grape Pomace Extract (RGPE) was proposed as a new active food packaging material. Pressurized Liquid Extraction (PLE) and Enhanced Solvent Extraction (ESE) techniques were employed to obtain the bioactive RGPE. Afterward the supercritical solvent impregnation conditions to obtain RGPE-natural jute fibers were studied, by varying pressure, modifier percentage and dried RGPE mass. PLE technique offered the highest bioactive extract at 20 MPa, 55 °C, 1 h residence time using C2H5OH:H2O (1:1 v/v), providing an EC50 of 3.35 ± 0.25 and antibacterial capacity against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa (MIC of 12.0, 1.5 and 4.0 mg/mL RGPE respectively). The natural jute fibers impregnated with 3 mL of that RGPE (90 mg/mL) at 50 MPa and 55 °C generated the most efficient packing material with regards to its food preservation potential.

Usage of supercritical fluid techniques to obtain bioactive alkaloid-rich extracts from cherimoya peel and leaves: extract profiles and their correlation with antioxidant properties and acetylcholinesterase and α-glucosidase inhibitory activities.

The agroindustrial sector is highly concerned with regards to reducing the environmental impact of waste from pruning activities (leaves, branches and bark) and from food industry processes (peels and seeds). In this sense, the wastes generated by cherimoya cultivation and processing industries should be contemplated as a valuable source of biologically active compounds. In this work, we have studied the bioactivity of alkaloid-rich Annona cherimola Mill. extracts obtain by means of supercritical fluid extraction techniques. The extracts were obtained from the peel and leaves using the following optimal conditions: 100 bar of pressure, 75 °C and 15% methanol as co-solvent. High antioxidant capacity (5304.23 ± 73.60 to 21 705.20 ± 1069.31 µmol Trolox equivalent per 100 g), and acetylcholinesterase (IC50 = 87.69 ± 3.42 to 515.02 ± 29.25 µg mL-1) and α-glucosidase (IC50 = 1097.76 ± 121.12 to 3206.88 ± 97.06 µg mL-1) inhibitory activities were exhibited by both peel and leaf extracts. Larger alkaloid contents were determined by UHPLC-ESI-MS analysis, with peel extracts presenting a high concentration of N-trans-feruloyl phenethylamine, while leaf extracts were rich in anonine. This work reports novel data on bioactivity of cherimoya peel and leaves and their potential as a source of bioactive compounds.

Long-Term Mangiferin Extract Treatment Improves Central Pathology and Cognitive Deficits in APP/PS1 Mice.

Alzheimer's disease (AD) is the most common cause of dementia; however, available treatments have had limited success. Therefore AD patients are in tremendous need of new pharmacological approaches that may delay or slow the progression of the disease. In addition to the classical neuropathological features, immunological and inflammatory processes are also involved in AD pathogenesis. Naturally occurring compounds, such as Mangifera indica Linn (MGF) extracts have previously been shown to significantly reduce peripheral inflammatory processes. In order to explore the role of MGF in AD central pathology, we have orally treated APP/PS1 mice for 22 weeks. While MGF did not affect amyloid pathology, tau hyperphosphorylation was significantly reduced in the cortex and hippocampus. Also, inflammatory processes, measured by microglia and astrocyte burdens, were diminished in MGF-treated mice. Moreover, neuronal morphological alterations, such as abnormal neurite curvature and dystrophies, highly increased in APP/PS1 mice, were significantly ameliorated by long-term MGF treatment. Reduction of all these pathological features were accompanied by compelling improvements of episodic and spatial memory in APP/PS1 mice treated with MGF. Altogether our data suggest that MGF may provide a useful tool to target different aspects of AD pathology and could lead to more effective future therapeutic or preventive strategies.

Mango leaf extract improves central pathology and cognitive impairment in a type 2 diabetes mouse model.

Epidemiological studies reveal that metabolic disorders, and specifically type 2 diabetes (T2D), are relevant risk factors to develop Alzheimer's disease (AD) and vascular dementia (VaD), the most common causes of dementia. AD patients are in a tremendous need of new therapeutic options because of the limited success of available treatments. Natural polyphenols, and concretely Mangifera indica Linn extract (MGF), have been reported to have antiinflammatory, antioxidant and antidiabetic activities. The role of MGF in central complications associated with T2D, after long-term treatment of db/db mice with MGF was analyzed. Metabolic parameters (body weight, glucose and insulin levels) as well as central complications including brain atrophy, inflammatory processes, spontaneous bleeding, tau phosphorylation and cognitive function in db/db mice treated with MGF for 22 weeks were assessed. MGF limits body weight gain in obese db/db mice. Insulin and C-peptide levels, indicative of pancreatic function, were longer maintained in MGF-treated animals. MGF reduced central inflammation by lowering microglia burden, both in the cortex and the hippocampus. Likewise, central spontaneous bleeding was significantly reduced in db/db mice. Cortical and hippocampal atrophy was reduced in db/db mice and tau hyperphosphorylation was lower after MGF treatment, resulting in partial recovery of learning and memory disabilities. Altogether, the data suggested that MGF treatment may provide a useful tool to target different aspects of AD and VaD pathology, and could lead to more effective clinical therapies for the prevention of metabolic related central complications associated with AD and VaD.

Helikaurolides A-D with a Diterpene-Sesquiterpene Skeleton from Supercritical Fluid Extracts of Helianthus annuus L. var. Arianna.

Four novel compounds (1-4) with an unprecedented skeleton that combines a sesquiterpene lactone and a kaurane diterpene acid were isolated from Helianthus annuus L. var. Arianna extract, which was obtained under supercritical conditions. The structures of 1-4 were elucidated by NMR and MS analyses. The biosynthetic routes involve sesquiterpene lactones and kauranic acid, both of which were previously isolated from this species.

Isolation of Bioactive Compounds from Sunflower Leaves (Helianthus annuus L.) Extracted with Supercritical Carbon Dioxide.

The work described herein is a continuation of our initial studies on the supercritical fluid extraction (SFE) with CO2 of bioactive substances from Helianthus annuus L. var. Arianna. The selected SFE extract showed high activity in the wheat coleoptile bioassay, in Petri dish phytotoxicity bioassays, and in the hydroponic culture of tomato seeds. Chromatographic fractionations of the extracts and a spectroscopic analysis of the isolated compounds showed 52 substances belonging to 10 different chemical classes, which were mainly sesquiterpene lactones, diterpenes, and flavonoids. Heliannuol M (31), helivypolides K and L (36, 37), and helieudesmanolide B (38) are described for the first time in the literature. Metabolites have been tested in the etiolated wheat coleoptile bioassay with good results in a noteworthy effect on germination. The most active compounds were also tested on tomato seeds, heliannuol A (30) and leptocarpin (45) being the most active, with values similar to those of the commercial herbicide.

Green extraction of antioxidants from different varieties of red grape pomace.

The extraction yield, phenolic content, anthocyanin content and antioxidant activity of extracts from different varieties of red grapes, Cabernet Sauvignon, Merlot, Petit Verdot, Syrah, Tempranillo and Tintilla, using pressurized green solvents have been analyzed. Two techniques were studied and compared: supercritical fluid extraction (SFE) with CO2 + 20% ethanol and pressurized liquid extraction (PLE) with either ethanol, water or an ethanol/water mixture as the extraction solvents. The Petit Verdot variety allowed the highest global and phenolic yield, and antioxidant activity. The best conditios for PLE obtained from the experimental design and kinetic study were 50% ethanol/water as the pressurized solvent at 90 bar, 120 °C, a flow rate of 5 g/min and, an extraction time of 90 min. A statistical analysis of variance has been performed and it was found that temperature is the only variable that has a statistical influence on the extraction yield. The antioxidant activity levels of the extracts are very promising and they are similar to those obtained with the antioxidant tocopherol.

SFE kinetics of bioactive compounds from Helianthus annuus L.

The kinetics of the supercritical fluid extraction of bioactive compounds from sunflower using CO(2) as solvent were studied in order to establish an efficient method for this extraction. The influence of time of extraction at different solvent flow rates was investigated. The extraction pressure and temperature were optimized in previous studies and these values used were 400 bar and 50 degrees C. The extraction yields and the bioactivity levels of the extracts were also analysed. The results indicate that the most appropriate extraction time is between 120 and 180 min depending on the solvent flow and the pretreatment of the sample. The dried sample extracted at 40 g/min, the congealed sample extracted at the same flow and 180 min, and the dried sample extracted at 25 g/min for 120 min showed the best activity profiles. The extractions were carried out in a pilot plant with an extraction vessel with a capacity of 2 L.

Supercritical CO(2) extraction of PAHs on spiked soil Co-solvent effect and solvent regeneration by ozonization.

The supercritical CO(2) extraction of four PAHs (acenaphthene, phenanthrene, anthracene and fluoranthene) from an artificially contaminated soil has been investigated. The effect of temperature (40-60 degrees C), pressure (300-500 bar) and extraction time (90-150 min) has been assessed by conducting a Box-Behnken experimental design. The results suggest the existence of perturbation variables other than the aforementioned controlled variables leading to a significant dispersion of extraction recoveries. With the exception of anthracene, an optimum in temperature (50 degrees C) is envisaged when extracting the PAHs. Analogously, with the exception of anthracene (positive effect), pressure does not have a significant influence. The recovery yield increases as extraction time is increased to a value of 120 min. No further improvement is experienced thereafter. If a co-solvent is used (H(2)O(2) aqueous solution) a beneficial effect can be noticed. Hydrogen peroxide concentration did exert no significant influence in the process. Methanol used to collect the extracted PAHs could be regenerated by gaseous ozone and reused in several consecutive runs.

Supercritical CO2 extraction of beta-carotene from a marine strain of the cyanobacterium Synechococcus species.

Dynamic extraction of carotenoids from a marine strain of Synechococcus sp. (Cyanophyceae) with supercritical CO2 (SC-CO2) was investigated with regard to operation pressure and temperature effects on extraction efficiency. Extraction yield (milligrams of pigment per gram of dry weight) for SC-CO2) was compared with the extraction yield for dimethylformamide (DMF). Carotenoids extracted with SC-CO2 were beta-carotene (Ct), zeaxanthin (Z), beta-cryptoxanthin (Cr), and equinenone; chlorophyll a was poorly extracted, whereas myxoxanthophyll, another major carotenoid, was not extracted under any experimental condition. The highest relative yield, which is defined here as y(r) = [(mg of pigment(SC-CO2)/mg of pigment(DMF))] x 100, was 76.1 +/- 8.6% for Ct, but it rose to 87.0 +/- 3.4% when 15% ethanol was used as cosolvent. The pressure effect on y(r) was found to be significant (p < 0.05) for both Cr and Z, along with total carotenoids, whereas the effect of square T (TT) was significant for only Ct. From empirical correlations, pairwise pressure (bar) and temperature (degrees C), respectively, for optimal extraction were determined to be (358, 50) for Ct, (454, 59) for Cr, and (500, 60) for Z. Cell disruption by sonication or detergent treatment of the biomass did not improve the extraction efficiency. Matrix structure together with material state could explain the low carotenoid extraction yield obtained with SC-CO2 as compared to DMF in Synechococcus sp. However, the process can be applied to selective extraction of different carotenoids.