Ephedra alata Seeds Confer Kidney Protection against Early Life Exposure to Acephate by Regulating Oxidative Insult and Activating Autophagy.

The aim of the current work was to examine for the first time the nephropreventive capacity of Ephedra alata seed extract (E) against maternal exposure to acephate in rat offspring. The in vivo results revealed that E. alata supplementation for 28 days (40 mg/kg b.w.) significantly attenuated the nephrotoxicity in adult offspring induced by acephate. In fact, it decreased the levels of creatinine and uric acid and increased the albumin content compared to the intoxicated group. The in utero studies showed that E. alata inhibited the renal oxidative stress generated by acephate exposure by reducing lipid peroxidation and enhancing antioxidant biomarker activities (GSH, CAT, and SOD). The inhibition of DNA fragmentation and the improvement of the ultrastructural changes highlighted the prophylactic effect of E. alata in renal tissue. Additionally, the immunofluorescence study showed the upregulation of LC3 gene expression, suggesting the capacity of E. alata extract to stimulate autophagic processes as a protective mechanism. Molecular docking analysis indicated that hexadecasphinganine, the major compound in E. alata, has a higher affinity toward the Na+/K+-ATPase, epithelial sodium channel (ENaC), and sodium hydrogen exchanger 3 (NHE3) genes than acephate. Hexadecasphinganine could be considered a potential inhibitor of the activity of these genes and therefore exerted its preventive capacity. The obtained findings confirmed that E. alata seed extract exerted nephropreventive capacities, which could be related to its bioactive compounds, which possess antioxidant activities.

Ephedra alata Subsp. Alenda as a Novel Source of Bioactive Phytochemicals: Characterization Based on the Mass Spectrometry and Profiling of Antioxidant and Anti-Inflammatory Properties.

The aim of the present study was to examine, for the first time, the phytochemical content of Ephedra alata pulp extract (EAP) and explore its antioxidant and anti-inflammatory capacities. High-performance liquid chromatography-electrospray ionization-quadrupole-time-of-flight mass spectrometry (HPLC-ESI-QTOF/MS) was used for phytochemical analysis and three in vitro antioxidant assays together with three in vitro anti-inflammatory tests were used for the assessment of biological activity. The HPLC-ESI-QTOF/MS analysis revealed the presence of 42 metabolites, including flavonoids, sphingolipides, fatty acids, ephedrine derivatives, and amino acid derivatives. In vitro findings revealed that EAP has interesting 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide, and ferrous ion chelating capacities (IC50 values were 0.57 mg/mL, 0.55 mg/mL, and 0.51 mg/mL for DPPH, superoxide radical, and ferrous ion, respectively). Furthermore, EAP showed a noticeable anti-inflammatory ability by inhibiting the two cyclooxygenase isoforms, COX-1 and COX-2 (IC50 of 59.1 and 58.8 µg/mL for COX-1 and COX-2, respectively), preventing protein denaturation (IC50 = 0.51 mg/mL), and protecting membrane stabilization (IC50 = 0.53 mg/mL). The results highlighted the use of Ephedra alata pulp as a potential source of natural compounds with therapeutic effects for the management of inflammatory disorders.

Grapevine Shoot Extract Rich in Trans-Resveratrol and Trans-ε-Viniferin: Evaluation of Their Potential Use for Cardiac Health.

A grapevine shoot extract (GSE) was obtained using ultrasound-assisted extraction and characterized. The main phenolic constituents were identified as stilbenoids. Among them, trans-resveratrol and trans-ε-viniferin stood out. The GSE was administered to an isoproterenol-induced myocardial injury animal model. The extract alleviated the associated symptoms of the administration of the drug, i.e., the plasma lipid profile was improved, while the disturbed plasma ion concentration, the cardiac dysfunction markers, the DNA laddering, and the necrosis of myocardial tissue were diminished. This effect could be related to the anti-oxidative potential of GSE associated with its antioxidant properties, the increased levels of endogenous antioxidants (glutathione and enzymatic antioxidants), and the diminished lipid peroxidative markers in the heart. The results also revealed angiotensin-converting enzyme (ACE)-inhibitory activity, which indicated the potential of GSE to deal with cardiovascular disease events. This work suggests that not only trans-resveratrol has a protective role in heart function but also GSE containing this biomolecule and derivatives. Therefore, GSE has the potential to be utilized in the creation of innovative functional ingredients.

Extraction Systems and Analytical Techniques for Food Phenolic Compounds: A Review.

Phenolic compounds are highly valuable food components due to their potential utilisation as natural bioactive and antioxidant molecules for the food, cosmetic, chemical, and pharmaceutical industries. For this purpose, the development and optimisation of efficient extraction methods is crucial to obtain phenolic-rich extracts and, for some applications, free of interfering compounds. It should be accompanied with robust analytical tools that enable the standardisation of phenolic-rich extracts for industrial applications. New methodologies based on both novel extraction and/or analysis are also implemented to characterise and elucidate novel chemical structures and to face safety, pharmacology, and toxicity issues related to phenolic compounds at the molecular level. Moreover, in combination with multivariate analysis, the extraction and analysis of phenolic compounds offer tools for plant chemotyping, food traceability and marker selection in omics studies. Therefore, this study reviews extraction techniques applied to recover phenolic compounds from foods and agri-food by-products, including liquid-liquid extraction, solid-liquid extraction assisted by intensification technologies, solid-phase extraction, and combined methods. It also provides an overview of the characterisation techniques, including UV-Vis, infra-red, nuclear magnetic resonance, mass spectrometry and others used in minor applications such as Raman spectroscopy and ion mobility spectrometry, coupled or not to chromatography. Overall, a wide range of methodologies are now available, which can be applied individually and combined to provide complementary results in the roadmap around the study of phenolic compounds.

Hepatopreventive properties of hydroxytyrosol and mannitol-rich extracts obtained from exhausted olive pomace using green extraction methods.

Exhausted olive pomace (EOP) is produced in olive-pomace oil extractors as a by-product. However, the obtention of bioactive compounds from EOP can reinsert it into the economy as a new bioresource before applying other exploitation ways. The objective of the present study was to investigate the phytochemical differences between aqueous and aqueous acetonic extracts from EOP (AE-EOP and AAE-EOP, respectively) obtained by hydrothermal and ultrasound-assisted extraction, respectively. The in vitro antioxidant activities and the in vivo hepatopreventive potential were evaluated. Using RP-HPLC-ESI-QTOF-MS, the chemical profile revealed that AE-EOP and AAE-EOP showed similar qualitative profiles, with some quantitative differences. Hydroxytyrosol and mannitol were the major compounds of the extracts. The investigation of antioxidant properties in vitro highlighted that AE-EOP was slightly more efficient in scavenging DPPH, ABTS, superoxide, and hydrogen peroxide radicals, when compared to AAE-EOP. Additionally, AE-EOP and AAE-EOP showed dose-dependent suppressive effects on pancreatic lipase activity. In vivo studies showed that AE-EOP and AAE-EOP presented interesting hepatopreventive capacities against CCl4 induced liver injury, as evidenced by (i) the preventive effects against DNA damage, (ii) the normalised hepatic biomarker parameters (ALT, AST, GGT, and LDH) and (iii) the normalised lipid profile (LDL-C, TC, TG, and HDL-C) through diminishing their levels, (iv) which was supported by Oil Red O analysis. Furthermore, AE-EOP and AAE-EOP reduced the oxidative stress in liver tissue by inhibiting lipid peroxidation together with the enhancement of the hepatic antioxidant activities (CAT, SOD and GPx) and GSH content. Additionally, AE-EOP and AAE-EOP exhibited an antifibrotic effect, which was clearly demonstrated by the histopathological examination using Picrosirius red staining. The obtained results support the use of EOP extracts in industries without further purification as antioxidants and against free radical induced damage.

Optimization of Microwave-Assisted Water Extraction to Obtain High Value-Added Compounds from Exhausted Olive Pomace in a Biorefinery Context.

Microwave-assisted water extraction (MAWE) was evaluated to obtain the valuable bioactive compounds hydroxytyrosol and mannitol from exhausted olive pomace (EOP). The influence of the operational parameters solid loading (3-15%, w/v), temperature (40-100 °C), and extraction time (4-40 min) was studied using an experimental design. The optimized conditions maximizing their joint extraction were 12% w/v solid loading, 100 °C temperature, and 16 min. It was possible to solubilize 5.87 mg of hydroxytyrosol/g EOP and 46.70 mg mannitol/g EOP. The extracts were also further characterized by liquid chromatography-mass spectrometry, which detected other hydroxytyrosol derivatives such as oleacein, verbascoside, and oleuropein. Moreover, the applied MAWE conditions promoted the co-extraction of proteinaceus material, which was also evaluated. In order to carry out an integral valorization of this waste, the extracted EOP solid was further evaluated chemically and microscopically before recovering the bioactive triterpenes. In particular, maslinic acid and oleanolic acid were obtained, 9.54 mg/g extracted solid and 3.60 mg/g extracted solid, respectively. Overall, MAWE can be applied as a first stage in the fractionation of EOP to support its valorization in a biorefinery framework.

A comparative study on the metabolites profiling of linseed cakes from Egyptian cultivars and antioxidant activity applying mass spectrometry-based analysis and chemometrics.

Linseed (Linum usitatissimum L.) is an economically important crop worldwide. After the oil extraction, the cake is generated as a by-product. This study addresses an untargeted profiling of the cakes of four Egyptian cultivars of linseed via RP-HPLC-DAD-QTOF-MS and MS/MS. Totally, 162 metabolites were detected in the samples, being classified into hydroxybenzoic acids (20), hydroxycinnamic acids (20), flavonoids (59), lignans (9), coumarins (1), amino acids (8), peptides (3), cyanogenic glycosides (6), jasmonates (15), nucleosides (2), organic acids (16), and sugars (3). Upon employing principal component analysis and hierarchical cluster analysis, the cultivars were adequately grouped, suggesting that this approach can serve for origin traceability. Besides, the total phenol content and antioxidant activity were determined and in the same line with the previous results. As far as we know, this is the first comprehensive metabolic profiling of linseed cakes that gives a prospect of its relevance for the food industry.

Sequential Extraction of Hydroxytyrosol, Mannitol and Triterpenic Acids Using a Green Optimized Procedure Based on Ultrasound.

Olive-derived biomasses contain bioactive compounds with health promoting effects as well as antioxidant and sweet-tasting properties. However, their sequential extraction has not been attained. In the present study, firstly antioxidants and mannitol were extracted from exhausted olive pomace (EOP) by an eco-friendly method, ultrasound-assisted water extraction (UAWE). The amplitude (20-80%), extraction time (2-18 min) and solid loading (2-15%, w/v) were evaluated according to a Box-Behnken experimental design. Using the response surface methodology, the optimal conditions for extraction were obtained: 80% amplitude, 11.5% solid loading and 16 min. It enabled the multi-response optimization of the total phenolic content (TPC) (40.04 mg/g EOP), hydroxytyrosol content (6.42 mg/g EOP), mannitol content (50.92 mg/g EOP) and antioxidant activity (ferric reducing power or FRAP, 50.95 mg/g EOP; ABTS, 100.64 mg/g EOP). Moreover, the phenolic profile of the extracts was determined by liquid chromatography-UV and mass spectrometry, identifying hydroxytyrosol as the main phenolic compound and other minor derivatives could be characterized. Scanning electron microscopy was used to analyze the morphological changes produced in the cellular structure of EOP after UAWE. In addition, the chemical composition of the extracted EOP solid was characterized for further valorization. Then, a second extraction step was performed in order to extract bioactive triterpenes from the latter solid. The triterpenes content in the extract was determined and the effect of the previous UAWE step on the triterpenes extraction was evaluated. In this case, the use of ultrasound enhanced the extraction of maslinic acid and oleanolic acid from pelletized EOP with no milling requirement. Overall, UAWE can be applied to obtain antioxidant compounds and mannitol as first extraction step from pelletized EOP while supporting the subsequent recovery of triterpenic acids.

Recovery of Bioactive Compounds from Industrial Exhausted Olive Pomace through Ultrasound-Assisted Extraction.

Exhausted olive pomace (EOP) is the main agro-industrial waste of the olive pomace extracting industries. It contains phenolic compounds and mannitol, so the extraction of these bioactive compounds should be considered as a first valorization step, especially if EOP is used as biofuel. Therefore, EOP was subjected to bath-type ultrasound-assisted extraction (UAE), and the effects of the acetone concentration (20-80%, v/v), solid load (2-15%, w/v), and extraction time (10-60 min) on the extraction of antioxidant compounds were evaluated according to a Box-Behnken experimental design. By means of the response surface methodology, the optimum conditions were obtained: 40% acetone, 8.6% solids, and 43 min. For all the extracts, the total phenolic content (TPC), flavonoid content (TFC), and antioxidant activity (DPPH, ABTS, and FRAP) were determined. With the aim of shortening the extraction time, a two-level factorial experiment design was also carried out using a probe-type UAE, keeping the solid load at 8.6% (w/v) and the acetone concentration at 40% (v/v), while the amplitude (30-70%) and the extraction time (2-12 min) were varied to maximize the aforementioned parameters. Finally, a maximum of phenolic compounds was reached (45.41 mg GAE/g EOP) at 12 min and 70% amplitude. It was comparable to that value obtained in the ultrasonic bath (42.05 mg GAE/g EOP), but, remarkably, the extraction time was shortened, which translates into lower costs at industrial scale. Moreover, the bioactive compound hydroxytyrosol was found to be the major phenolic compound in the extract, i.e., 5.16 mg/g EOP (bath-type UAE) and 4.96 mg/g EOP (probe-type UAE). Other minor phenolic compounds could be detected by capillary zone electrophoresis and liquid-chromatography-mass spectrometry. The sugar alcohol mannitol, another bioactive compound, was also found in the extract, and its content was determined. Thus, the use of this technology can support the valorization of this waste to obtain bioactive compounds, including mannitol, hydroxytyrosol, and other derivatives, before being applied for other uses.

HPLC-ESI-QTOF-MS/MS profiling and therapeutic effects of Schinus terebinthifolius and Schinus molle fruits: investigation of their antioxidant, antidiabetic, anti-inflammatory and antinociceptive properties.

The aim of the current work was to study the phytochemical variability among Schinus terebinthifolius (STE) and Schinus molle (SME) fruit extracts. The in vitro antioxidant, antihemolytic, antidiabetic, and macromolecule damage protective activities, as well as, the in vivo anti-inflammatory and antinociceptive capacities were assessed. Using the HPLC-ESI-QTOF/MS analysis, the chemical profile of fruit extract varied between S. terebinthifolius (30 compounds) and S. molle (16 compounds). The major compound was masazino-flavanone (5774.98 and 1177.65 µg/g sample for STE and SME, respectively). The investigations highlighted significant antioxidant proprieties when using ABTS radical (IC50; 0.12 and 0.14 mg/ml for STE and SME, respectively), superoxide (IC50; 0.17 and 0.22 mg/ml for STE and SME, respectively) and hydrogen peroxide (IC50; 014 and 0.17 mg/ml for STE and SME, respectively). In addition, STE and SME proved preventive effects against H2O2-induced hemolysis (IC50; 0.22 and 0.14 mg/ml for STE and SME, respectively). The in vitro antidiabetic effect revealed that STE and SME exhibited important inhibitory effects against α-amylase (IC50; 0.13 and 0.19 mg/ml for STE and SME, respectively) and α-glycosidase (IC50; 0.21 and 0.18 mg/ml for STE and SME, respectively) when compared with acarbose. Furthermore, the extracts showed potent inhibitory activity against AAPH-induced plasmid DNA damage, and protein oxidation. In vivo study revealed that STE and SME presented interesting antinociceptive and anti-inflammatory capacities. All observed effects highlighted the potential application of Schinus fruit extract in food and pharmaceutical industries against ROS-induced damage.

Metabolic Profiling of the Oil of Sesame of the Egyptian Cultivar 'Giza 32' Employing LC-MS and Tandem MS-Based Untargeted Method.

Sesame (Sesamum indicum L.) is a global oil crop. Sesame oil has been regarded as functional oil with antioxidant properties in several in vivo studies but little is known about its minor fraction. In this line, this study figures out the profile of the polar fraction of Egyptian cultivar Giza 32 sesame oil (SG32 oil) employing reversed-phase high-performance liquid chromatography coupled with diode array detection and electrospray ionization-quadrupole-time-of-flight-mass spectrometry and tandem MS. The characterization of the sesame oil metabolites depended on the observation of their retention time values, accurate MS, and MS/MS data, with UV spectra, and compared with relevant literature and available standards. Remarkably, 86 metabolites were characterized and sub-grouped into phenolic acids, lignans, flavonoids, nitrogenous compounds, and organic acids. From the characterized metabolites, 72 compounds were previously characterized in SG32 cake, which presented antioxidant properties, and hence it could contribute to SG32 oil antioxidant properties. Further studies are required to state the presence of such phenolics in commercial sesame oils and what of these compounds resist oil refining.

Olive Pomace-Derived Biomasses Fractionation through a Two-Step Extraction Based on the Use of Ultrasounds: Chemical Characteristics.

Olive-derived biomass is not only a renewable bioenergy resource but also it can be a source of bioproducts, including antioxidants. In this study, the antioxidant composition of extracted olive pomace (EOP) and a new byproduct, the residual fraction from olive pit cleaning (RFOPC or residual pulp) was characterized and compared to olive leafy biomass, which have been extensively studied as a source of antioxidants and other bioactive compounds with pharmacological properties. The chemical characterization showed that these byproducts contain a high amount of extractives; in the case of EOP, it was even higher (52.9%) than in olive leaves (OL) and olive mill leaves (OML) (35.8-45.1%). Then, ultrasound-assisted extraction (UAE) was applied to recover antioxidants from the extractive fraction of these biomasses. The solubilization of antioxidants was much higher for EOP, correlating well with the extractives content and the total extraction yield. Accordingly, this also affected the phenolic richness of the extracts and the differences between all biomasses were diminished. In any case, the phenolic profile and the hydroxytyrosol cluster were different. While OL, OML, and EOP contained mainly hydroxytyrosol derivatives and flavones, RFOPC presented novel trilignols. Other compounds were also characterized, including secoiridoids, hydroxylated fatty acids, triterpenoids, among others, depending on the bioresource. Moreover, after the UAE extraction step, alkaline extraction was applied recovering a liquid and a solid fraction. While the solid fraction could of interest for further valorization as a biofuel, the liquid fraction contained proteins, sugars, and soluble lignin, which conferred antioxidant properties to these extracts, and whose content depended on the biomass and conditions applied.

Schinus terebinthifolius fruits intake ameliorates metabolic disorders, inflammation, oxidative stress, and related vascular dysfunction, in atherogenic diet-induced obese rats. Insight of their chemical characterization using HPLC-ESI-QTOF-MS/MS.

ETHNOPHARMACOLOGY RELEVANCE: Schinus terebinthifolius is traditionally used for its anti inflammatory capacity, and indicated as a cardioprotective agent, whereas, its preventive effect against atherogenic diet fed (AD) induced metabolic disorders and the underlying mechanisms has not yet been explored. AIM OF THE STUDY: This study was undertaken to investigate the ameliorative role of Schinus terebinthifolius fruits extract (STFE) against cardiovascular problem, oxidative and inflammatory status related to obesity in rats fed an atherogenic diet. MATERIALS AND METHODS: The metabolites profile in STFE was evaluated using HPLC-DAD-ESI-QTOF-MS/MS analysis. In Wistar rats, atherogenic diet was added for 9 weeks to induce lipid accumulation simultaneously with STFE (50 mg/kg b. w) or saline treatment. Biochemical, oxidant, and inflammatory criteria together with hepatic and arterial integrity examination were assessed. RESULTS: A total of thirty three metabolites were identified using HPLC-DAD-ESI-QTOF-MS, among them masazino-flavanone was the major compound (2645.50 µg/g DW). The results indicated that STFE supplementation during 9 weeks (50 mg/kg b. w.) significantly attenuated the altered lipid profile by decreasing the levels of TC, TG, LDL-C and increasing the HDL-C content both in plasma and liver, when compared with the AD-group. The histological analysis using ORO staining revealed a decrease in the lipid droplet deposit in the cytoplasm of hepatocytes of STFE + AD group. The addition of STFE could improve the glycemic status of AD-treated rats by decreasing the glucose and insulin secretion, and ameliorating the hepatic glycogen synthesis. The harmful effects of atherogenic diet on hepatic oxidative stress indicators (MDA, PC, GSH, SOD, CAT, and GPx), biochemical markers (AST, ALT, LDH and ALP), and liver function, were found to be decreased by the addition of STFE. Moreover, the reduction of inflammatory markers (CRP, IL-6 and TNF-α), associated to alleviating of aortic oxidative stress and integrity, highlighted the positive anti-atherogenic effect of STFE. CONCLUSION: Overall, the pleiotropic protective effect observed with S. terebinthifolius fruits might be related to the presence of various bioactive compounds.

Integrated Profiling of Fatty Acids, Sterols and Phenolic Compounds in Tree and Herbaceous Peony Seed Oils: Marker Screening for New Resources of Vegetable Oil.

Tree peonies (Paeonia ostii and Paeonia rockii) are popular ornamental plants. Moreover, these plants have become oil crops in recent years. However, there are limited compositional studies focused on fatty acids. Therefore, this work aims to reveal compositional characteristics, regarding fatty acids, sterols, γ-tocopherol and phenolic compounds, of tree peony seed oils from all major cultivation areas in China, and to compare with herbaceous peony seed oil. For that, an integrative analysis was performed by GC-FID, GC-MS and UHPLC-Q-TOF-MS technologies. The main fatty acid was α-linolenic acid (39.0-48.3%), while ß-sitosterol (1802.5-2793.7 mg/kg) and fucosterol (682.2-1225.1 mg/kg) were the dominant phytosterols. Importantly, 34 phenolic compounds, including paeonol and "Paeonia glycosides" (36.62-103.17 µg/g), were characterized in vegetable oils for the first time. Conclusively, this work gives new insights into the phytochemical composition of peony seed oil and reveals the presence of bioactive compounds, including "Paeonia glycosides".

Extraction Strategies to Recover Bioactive Compounds, Incorporation into Food and Health Benefits: Current Works and Future Challenges.

There are numerous studies in the literature about bioactive products (extracts, essential oils, oleoresins, hydrolysates, etc [...].

Content of phenolic compounds and mannitol in olive leaves extracts from six Spanish cultivars: Extraction with the Soxhlet method and pressurized liquids.

Olive leaves are considered a promising source of bioactives such as phenolic compounds and mannitol. The extraction of high added value products is an issue of great interest and importance from the point of view of their exploitation. However, the content of these compounds can differ between cultivars and extraction methods. In this work, six olive leaves cultivars, including three wild cultivars, and two extraction processes (an innovative and alternative technique, pressurized liquid extraction, and a conventional Soxhlet extraction) were evaluated and compared towards the selective recovery of bioactive compounds. The wild cultivars showed the highest content of phenolic and flavonoid compounds, being oleuropein the compound present in higher amount. Findings also revealed that the highest mannitol content in the extracts was observed with the commercial cultivars, specifically in Arbequina. It is thus possible to decide which cultivars to use in order to obtain the highest yield of each bioproduct.

Zygophyllum album saponins prevent atherogenic effect induced by deltamethrin via attenuating arterial accumulation of native and oxidized LDL in rats.

The current study aimed to examine, for the first time, the relationship between exposure to deltamethrin (DLM) and atherogenic lipid profile disorders in adult Wistar rats, as well as, to verify the mechanism of the beneficial role of Zygophyllum album leaves extracts (ZALE). The experimental study was assessed using DLM (4 mg/kg b.w) either alone or co administered with ZALE (400 mg/kg b.w) orally for 90 days in rats. RP-HPLC-DAD-ESI-QTOF-MS was used to identify the bioactive metabolites present in ZALE. Plasmatic and aortic total cholesterol (TC), LDL-cholesterol (LDL-C), native LDL (LDL-apo B-100) and oxidized LDL (ox-LDL) were evaluated using auto-analyzer and a sandwich ELISA, respectively. The protein expressions of LDLR (native LDL receptor) and CD36 (Scavenger receptor class B) were evaluated in aorta or liver with a Western blot. The pathology has been confirmed with lipid stain (Oil Red O). Phytochemicals analysis revealed the presence of fifteen saponins in ZALE. Rats intoxicated with DLM revealed a significant increase in plasmatic and aortic lipid profile (TC, LDL-C, LDL-apo B-100 and ox-LDL), as well as, the concentration of the plasmatic cytokines include TNF-α, IL-2 and IL-6, compared to control. Hepatic native LDL and aortic CD36 receptor expression were increased in DLM treated group, however aortic LDL-R does not present any modification, when compared to control. The detected disturbances in lipid parameters were supported by Oil Red O applied. Due to their antioxidant activity, the bioactive compounds in ZALE as powerful agents able to prevent the pro-atherogenic effect observed in DLM-treated animals. These metabolites modulated most of inflammatory markers, prevented accumulation of lipid and lipoprotein biomarkers, regulated the major receptor regulators of hepatic cholesterol metabolism, as well as normalize lipid distribution in liver and aorta tissue.

Valorization of olive mill leaves through ultrasound-assisted extraction.

Olive leaves farmed from trees are valuable for the production of functional extracts. Nonetheless, olive leaves (containing thin branches), which are separated during olives cleaning in the mill, have received little attention. In this context, a multiple response optimization was performed to maximize at once the yield, total phenolic content, oleouropein and antioxidant activity obtained by ultrasound-assisted extraction of this low-cost byproduct. The optimum was achieved using the following operational parameters: solid-to-liquid ratio, 5.9%; ethanol concentration, 47%; extraction time, 50 min. This enabled to obtain an extract with both high level of oleuropein and antioxidant activity. Besides oleuropein, other minor phenolic compounds were characterized in the extract, which could contribute to the antioxidant activity as Pearson correlation suggested. After this extraction step, how the phenolic extraction affects the recovery/profile of other constituents was evaluated, looking for the integral valorization of this resource towards the zero-waste.

HPLC-DAD-ESI-QTOF-MS/MS profiling of Zygophyllum album roots extract and assessment of its cardioprotective effect against deltamethrin-induced myocardial injuries in rat, by suppression of oxidative stress-related inflammation and apoptosis via NF-κB signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Zygophyllum album is widely used to treat many cardiovascular diseases (CVDs) and as anti-inflammatory plant. AIM OF THE STUDY: This study aimed to investigate the mechanism of the potential protective effects of Zygophyllum album roots extract (ZARE) against myocardial damage and fibrosis induced by a chronic exposure to deltamethrin (DLM) in rats. MATERIALS AND METHODS: Bioactive compounds present in ZARE were analyzed by HPLC-DAD-ESI-QTOF-MS/MS. In vivo, DLM (4 mg/kg body weight), ZARE (400 mg/kg body weight) and DLM with ZARE were administered to rats orally for 60 days. Biochemical markers (LDH, ALT, CK, CK-MB and cTn-I) were assessed in the plasma by an auto-analyzer. Pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-6) were evaluated by a sandwich ELISA. NF-κB was quantified at mRNA levels by real time PCR. Heart tissue was used to determine cardiac oxidative stress markers (MDA, PC, SOD, CAT, and GPx). Masson's Trichrome (MT) and Sirius Red (SR) stainings were used for explored fibrosis statues. RESULTS: Phytochemical analysis using HPLC-DAD-ESI-QTOF-MS/MS revealed the presence of twenty six molecules including phenolic compounds and saponins. ZARE significantly improved the heart injury markers (LDH, ALT, CK, CK-MB and cTn-I), lipid peroxidation (MDA), protein oxidation (PC), antioxidant capacity (SOD, CAT, and GPx), and DNA structure, which were altered by DLM exposure. Moreover, ZARE cotreatment reduced the expressions of NF-κB, decreased plasmatic pro-inflammatory cytokines concentration (TNF-α, IL-1ß and IL-6), and suppressed the myocardial collagen deposition, as observed by Sirius Red and Masson's Trichrome staining. CONCLUSION: ZARE ameliorated the severity of DLM-induced myocardial injuries through improving the oxidative status and reducing profibrotic cytokines production. The ZARE actions could be mediated by downregulation of NF-κB mRNA.

Integrated Process for Sequential Extraction of Bioactive Phenolic Compounds and Proteins from Mill and Field Olive Leaves and Effects on the Lignocellulosic Profile.

The extraction of bioactive compounds in a biorefinery context could be a way to valorize agri-food byproducts, but there is a remaining part that also requires attention. Therefore, in this work the integrated extraction of phenolic compounds, including the bioactive oleuropein, and proteins from olive mill leaves was addressed following three schemes, including the use of ultrasound. This affected the total phenolic content (4475.5-6166.9 mg gallic acid equivalents/100 g), oleuropein content (675.3-1790.0 mg/100 g), and antioxidant activity (18,234.3-25,459.0 µmol trolox equivalents/100 g). No effect was observed on either the protein recovery or the content of sugars and lignin in the extraction residues. Concerning the recovery of proteins, three operational parameters were evaluated by response surface methodology. The optimum (63.1%) was achieved using NaOH 0.7 M at 100 °C for 240 min. Then, the selected scheme was applied to olive leaves from the field, observing differences in the content of some of the studied components. It also changed the lignocellulosic profile of the extraction residues of both leaf types, which were enriched in cellulose. Overall, these results could be useful to diversify the valorization chain in the olive sector.