Leonurine Preconditioning Attenuates Ischemic Acute Kidney Injury in Rats by Promoting Nrf2 Nuclear Translocation and Suppressing TLR4/NF-κB Pathway.

Despite the precise mechanisms for renal ischemia/reperfusion (I/R)-induced acute kidney injury (AKI) are poorly understood, nuclear factor erythroid 2 related factor 2 (Nrf2) and Toll-like receptor 4 (TLR4) pathways were considered as the important targets. Leonurine (LEO) is a special alkaloid extracted from Chinese motherwort (Leonurus japonicus Houtt), which has an anti-inflammatory effect and reduces oxidative stress. We conducted the study to explore the efficacy of LEO against I/R-induced AKI in rats and further investigated the underlying mechanisms. Ischemic renal injury was induced by temporary vascular clamping for 45 min. We have measured the levels of inflammation-related biomarkers and antioxidative stress markers. Next, Western blot analysis and Real-time PCR were performed to analyze whether the Nrf2 and TLR4/nuclear factor-kappaB (NF-κB) pathways were involved in this process. We found that LEO pretreatment remarkably decreased serum creatinine and blood urea nitrogen (BUN) in I/R rats and attenuated acute tubular damage. In addition, LEO markedly increased the expression of antioxidant proteins and decreased the levels of inflammatory factors. Further study revealed that LEO promoted Nrf2 into the nucleus, promoted the expression of heme oxygenase-1 (HO-1) and quinone oxidoreductase 1 (NQO-1), and suppressed the TLR4/NF-κB signal pathway in kidney tissues of ischemic AKI rats. The study reveals that LEO has a protective effect to prevent ischemic AKI through activation of Nrf2 nuclear translocation resisting oxidative stress injury and inhibition of the TLR4/NF-κB pathway mediated inflammatory gene expression.

Stepwise tracking strategy to screen ingredient from Galla Chinensis based on the "mass spectrometry guided preparative chromatography coupled with systems pharmacology".

ETHNOPHARMACOLOGICAL RELEVANCE: Galla chinensis, a traditional Chinese herbal medicine, was widely used to treat ulcerative colitis (UC) in folk prescriptions, however, its active ingredients and mechanism of action in the treatment of UC remain unclear. AIM OF THE STUDY: The aim of our study was to discover the lead compounds and anti-inflammatory active ingredients of Galla chinensis and clarify their molecular mechanism for UC treatment. MATERIALS AND METHODS: The ingredients of Galla chinensis were prepared by column and mass spectrometry guided preparative chromatography. Besides, the relationship among the ingredients of Galla chinensis and targets was predicted by systems pharmacology. Additionally, Lipopolysaccharide (LPS)-induced RAW264.7 macrophages were used as in vitro model. The cell viability, the level of the pro-inflammatory factors, the generation of reactive oxygen species (ROS), and trans epithelial electric resistance (TEER) values were detected to screen out the active ingredients of Galla chinensis. Moreover, 4% dextran sodium sulfate (DSS)-induced ulcerative colitis mice were used as the UC animal model. The disease activity index (DAI), pathological degree of colon tissue, activities of antioxidant-related enzymes and expression level of pro-inflammatory cytokines were performed to assess the anti-UC effects of the active ingredients. Meanwhile, the mRNA expression level of inflammatory factors and antioxidant related genes were analyzed by real-time quantitative polymerase chain reaction (Q-PCR). And the expression of nuclear factor erythroid-2 related factor 2 (Nrf2) pathway related proteins, intestinal mucosal proteins and nuclear factor kappa-B (NF-κB) pathway related proteins in colon tissues were analyzed by Western Blotting. RESULTS: Herein, a stepwise tracking strategy was adopted to screen out the anti-inflammatory active ingredients of Galla Chinensis based on "preparative chromatography pharmacology combined with mass spectrometry guidance and system". 11 categories of ingredients of Galla chinensis were prepared and ethyl gallate (EG) was screened out the lead compound and anti-inflammatory active ingredient of Galla Chinensis through in silico, in vitro and in vivo studies. In addition, EG had a significant therapeutic effect on ameliorating DSS-induced UC mice and protected intestinal mucosal integrity through Nrf2 and NF-κB signaling pathway. CONCLUSION: Ethyl gallate was the lead compound and anti-inflammatory active ingredient in Galla chinensis. And it was discovered for the first time that EG could treat mice with ulcerative colitis. This research not only found the lead compound of Galla Chinensis for UC treatment and determined the possible mechanism, but also provided valuable references for finding lead compounds from natural products by systems pharmacology coupled with equivalent components group technology.

Optimized extraction of polyphenolic antioxidants from the leaves of Himalayan Oak species.

In this study heat-assisted extraction conditions were optimized to enhance extraction yield of antioxidant polyphenols from leaves of Himalayan Quercus species. In initial experiments, a five-factor Plackett-Burman design including 12 experimental runs was tested against the total polyphenolic content (TP). Amongst, XA: extraction temperature, XC: solvent concentration and XE: sample-to-solvent ratio had shown significant influence on yield. These influential factors were further subject to a three-factor-three-level Box-Wilson Central Composite Design; including 20 experimental runs and 3D response surface methodology plots were used to determine optimum conditions [i.e. XA: (80°C), XC:(87%), XE: (1g/40ml)].This optimized condition was further used in other Quercus species of western Himalaya, India. The High-Performance Liquid Chromatography (HPLC) revealed occurrence of 12 polyphenols in six screened Quercus species with the highest concentration of catechin followed by gallic acid. Amongest, Q. franchetii and Q. serrata shared maximum numbers of polyphenolic antioxidants (8 in each). This optimized extraction condition of Quercus species can be utilized for precise quantification of polyphenols and their use in pharmaceutical industries as a potential substitute of synthetic polyphenols.

Characterization and Identification of Bioactive Polyphenols in the Trapabispinosa Roxb. Pericarp Extract.

In this study, we present the isolation and characterization of the structure of six gallotannins (1-6), three ellagitannins (7-9), a neolignan glucoside (10), and three related polyphenolic compounds (gallic acid, 11 and 12) from Trapa bispinosa Roxb. pericarp extract (TBE). Among the isolates, the structure of compound 10 possessing a previously unclear absolute configuration was unambiguously determined through nuclear magnetic resonance and circular dichroism analyses. The α-glucosidase activity and glycation inhibitory effects of the isolates were evaluated. Decarboxylated rugosin A (8) showed an α-glucosidase inhibitory activity, while hydrolyzable tannins revealed stronger antiglycation activity than that of the positive control. Furthermore, the identification and quantification of the TBE polyphenols were investigated by high-performance liquid chromatography coupled to ultraviolet detection and electrospray ionization mass spectrometry analysis, indicating the predominance of gallic acid, ellagic acid, and galloyl glucoses showing marked antiglycation properties. These findings suggest that there is a potential food industry application of polyphenols in TBE as a functional food with antidiabetic and antiglycation activities.

Pimenta dioica (L.) Merr. Bioactive Constituents Exert Anti-SARS-CoV-2 and Anti-Inflammatory Activities: Molecular Docking and Dynamics, In Vitro, and In Vivo Studies.

In response to the urgent need to control Coronavirus disease 19 (COVID-19), this study aims to explore potential anti-SARS-CoV-2 agents from natural sources. Moreover, cytokine immunological responses to the viral infection could lead to acute respiratory distress which is considered a critical and life-threatening complication associated with the infection. Therefore, the anti-viral and anti-inflammatory agents can be key to the management of patients with COVID-19. Four bioactive compounds, namely ferulic acid 1, rutin 2, gallic acid 3, and chlorogenic acid 4 were isolated from the leaves of Pimenta dioica (L.) Merr (ethyl acetate extract) and identified using spectroscopic evidence. Furthermore, molecular docking and dynamics simulations were performed for the isolated and identified compounds (1-4) against SARS-CoV-2 main protease (Mpro) as a proposed mechanism of action. Furthermore, all compounds were tested for their half-maximal cytotoxicity (CC50) and SARS-CoV-2 inhibitory concentrations (IC50). Additionally, lung toxicity was induced in rats by mercuric chloride and the effects of treatment with P. dioca aqueous extract, ferulic acid 1, rutin 2, gallic acid 3, and chlorogenic acid 4 were recorded through measuring TNF-α, IL-1ß, IL-2, IL-10, G-CSF, and genetic expression of miRNA 21-3P and miRNA-155 levels to assess their anti-inflammatory effects essential for COVID-19 patients. Interestingly, rutin 2, gallic acid 3, and chlorogenic acid 4 showed remarkable anti-SARS-CoV-2 activities with IC50 values of 31 µg/mL, 108 µg/mL, and 360 µg/mL, respectively. Moreover, the anti-inflammatory effects were found to be better in ferulic acid 1 and rutin 2 treatments. Our results could be promising for more advanced preclinical and clinical studies especially on rutin 2 either alone or in combination with other isolates for COVID-19 management.

Anti-damage effect of theaflavin-3'-gallate from black tea on UVB-irradiated HaCaT cells by photoprotection and maintaining cell homeostasis.

Keratinocytes are rich in lipids and are the main sensitive cells to ultraviolet (UV) rays. Theaflavins are the core functional components of black tea and are known as the "soft gold" in tea. In this study, ultraviolet-B (UVB) irradiation caused apoptosis and necrosis of human epidermal keratinocytes (HaCaT). EGCG and the four theaflavins had anti-UVB damage activity, among which theaflavin-3'-gallate (TF3'G) had the best activity. The results of biophysical and molecular biology experiments showed that TF3'G has anti-damage effects on UVB-irradiated HaCaT cells through the dual effects of photoprotection and maintenance of cell homeostasis. That is, TF3'G preincubation could absorb UV rays, reduce the accumulation of aging-related heterochromatin (SAHF) formation, increase mitochondrial membrane potential, downregulate NF-κB inflammation pathways, inhibit the formation of cytotoxic aggregates, and protect biological macromolecules Structure, etc. The accumulation of conjugated π bonds and the balance benzoquinone are the core functional structure of TF3'G with high efficiency and low toxicity. The study indicates that TF3'G has the potential to inhibit the photoaging and intrinsic aging of skin cells.

Changes in the Antioxidative Activity and the Content of Phenolics and Iridoids during Fermentation and Aging of Natural Fruit Meads.

The aim of the study was to investigate changes in the content of biologically active compounds during the fermentation and aging of natural meads with the addition of three Cornelian cherry juices from three cultivars: 'Koralovyi', 'Podolski' and 'Yantarnyi', in the amount of 10% v/v. After the fermentation process the content of gallic and ellagic acids significantly increased, in relation to wort. Whereas the greatest losses were observed among unstable anthocyanins. The three-month aging process also reduced the content of the analyzed compounds except for ellagic acid, the content of which increased by up to 90%. The content of biologically active compounds, including iridoids and antioxidant phenolics, are constantly changing in the process of fermentation and aging of fruit meads. The studies proved that the addition of Cornelian cherry juice allows significantly enriched classic meads with new biologically active compounds, such as: exceptional iridoids (loganic acid, cornuside, loganine, sweroside), flavonols, phenolic acids and anthocyanins.

Preparative isolation of antioxidative gallic acid derivatives from Saxifraga tangutica using a class separation method based on medium-pressure liquid chromatography and reversed-phase liquid chromatography.

Saxifraga tangutica is widely used as a medicinal herb to treat hepatic diseases. Here, we developed a class separation method to separate gallic acid derivatives 1,1-diphenyl-2-picrylhydrazyl inhibitors from the methanol extract of Saxifraga tangutica. Firstly, an MCI GEL CHP20P medium-pressure liquid chromatography was used to pretreat the crude extract from Saxifraga tangutica (500 g) and the target sample (fraction Fr1, 1.7 g) was obtained. Then, an online reversed-phase liquid chromatography-1,1-diphenyl-2-picrylhydrazyl assay was employed for recognizing potential 1,1-diphenyl-2-picrylhydrazyl inhibitors and six 1,1-diphenyl-2-picrylhydrazyl inhibitors fractions were recognized from fraction Fr1. Subsequently, the six 1,1-diphenyl-2-picrylhydrazyl inhibitors fractions were isolated via a ReproSil-Pur C18 AQ preparative column. During the separation process, the hydrophilic liquid chromatography was used to enrich the target compounds (Fr1-3-1-1 and Fr1-3-1-2) from the fraction Fr1-3, which were hardly isolated only by one step reversed-phase liquid chromatography. Finally, six gallic acid derivatives were obtained and identified as gallic acid (Fr1-1-1), gallic acid 3-O-ß-D-glucoside (Fr1-1-2), protocatechuic acid (Fr1-2), 4-O-galloyl-(-)-shikimic acid (Fr1-3-1-1), 5-O-galloyl-(-)-shikimic acid (Fr1-3-1-2), and 3-O-galloyl-shikimic acid (Fr1-4), respectively. Thus, the present study indicated that this method was highly efficient for the preparative separation of gallic acid derivatives 1,1-diphenyl-2-picrylhydrazyl inhibitors from natural products.

Phenolic Profiles of Ten Australian Faba Bean Varieties.

Although Australia is the largest exporter of faba bean globally, there is limited information available on the levels of bioactive compounds found in current commercial faba bean varieties grown in this country. This study profiled the phenolic acid and flavonoid composition of 10 Australian faba bean varieties, grown at two different locations. Phenolic profiling by HPLC-DAD revealed the most abundant flavonoid to be catechin, followed by rutin. For the phenolic acids, syringic acid was found in high concentrations (72.4-122.5 mg/kg), while protocatechuic, vanillic, p-hydroxybenzoic, chlorogenic, p-coumaric, and trans-ferulic acid were all found in low concentrations. The content of most individual phenolics varied significantly with the variety, while some effect of the growing location was also observed. This information could be used by food processors and plant breeders to maximise the potential health benefits of Australian-grown faba bean.

Phenolic Compounds, Antioxidant Activity and Fatty Acid Composition of Roasted Alyanak Apricot Kernel.

The oil recovery from Alyanak apricot kernel was 36.65% in control (unroasted) and increased to 43.77% in microwave-roasted kernels. The total phenolic contents in extracts from apricot kernel were between 0.06 (oven-roasted) and 0.20 mg GAE/100 g (microwave-roasted) while the antioxidant activity varied between 2.55 (oven-roasted) and 19.34% (microwave-roasted). Gallic acid, 3,4-dihydroxybenzoic acid, (+)-catechin and 1,2-dihydroxybenzene were detected as the key phenolic constituents in apricot kernels. Gallic acid contents varied between 0.53 (control) and 1.10 mg/100 g (microwave-roasted) and 3,4-dihydroxybenzoic acid contents were between 0.10 (control) and 0.35 mg/100 g (microwave-roasted). Among apricot oil fatty acids, palmitic acid contents ranged from 4.38 (oven-roasted) to 4.76% (microwave-roasted); oleic acid contents were between 65.73% (oven-roasted) and 66.15% (control) and linoleic acid contents varied between 26.55 (control) and 27.12% (oven-roasted).

Four New Gallate Derivatives from Wine-Processed Corni Fructus and Their Anti-Inflammatory Activities.

Four new gallate derivatives-ornusgallate A, ent-cornusgallate A, cornusgallate B and C (1a, 1b, 2, 3)-were isolated from the wine-processed fruit of Cornus officinalis. Among them, 1a and 1b are new natural compounds with novel skeletons. Their chemical structures were elucidated by comprehensive spectroscopy methods including NMR, IR, HRESIMS, UV, ECD spectra and single-crystal X-ray diffraction analysis. The in vitro anti-inflammatory activities of all compounds were assayed in RAW 264.7 cells by assessing LPS-induced NO production. As the result, all compounds exhibited anti-inflammatory activities at attested concentrations. Among the tested compounds, compound 2 exhibited the strongest anti- inflammatory activity.

Characterization of alcohol/salt aqueous two-phase system for optimal separation of gallic acids.

Gallic acid (GA) is a hydrophilic polyphenol which is noteworthy for strong antioxidant capacity. The drawbacks of conventional extraction approaches such as time-consuming and high processing cost are often viewed as a hurdle to extract GA from plant sources in industrial scale. Aqueous two-phase system (ATPS) is a separation approach which can be employed as an alternative to the conventional approaches. The partition behaviour of GA in an alcohol/salt ATPS was investigated in this study to aid the development of industrial scale ATPS to extract GA from natural sources. The separation of GA was characterized by determining the types of alcohol and salt, phase composition, sample load, pH of the system and addition of adjuvants applied in the alcohol/salt ATPS construction. The hydrophilic GA was targeted to the salt-rich phase of the alcohol/salt ATPS with a partition coefficient (KGA) of 25.00 ± 0.00. The optimum condition of ATPS for the maximum partition of GA was achieved in ATPS comprised of 24% (w/w) 1-propanol and 22% (w/w) phosphate salt at pH 8 with 5% (w/w) of 1 mg/mL sample loading and 2% (w/w) NaCl addition. The findings suggest that ATPS can be applied for separation of GA from various natural sources.

Bioassay-guided fractionation and identification of antidiabetic compounds from the rind of Punica Granatum Var. nana.

This study aimed to evaluate the antidiabetic potential of the rind of Punica granatum var. nana. Acute oral toxicity test revealed the safety profile of its ethanolic extract. The extract was administered at 200 mg/kg b.wt to streptozotocin-induced diabetic rats. Serum diagnostic markers of diabetes (insulin, glucose and glycated hemoglobin), inflammatory mediators (tumor necrosis factor-α, interleukin-6, and nitric oxide), and oxidative stress (total antioxidant capacity and reduced glutathione and malondialdehyde) were assayed. The ethanolic extract was further fractionated and assessed for the aforementioned bioactivities at two different doses (100 and 200 mg/kg b.wt). The results revealed that the ethyl acetate fraction of rind exhibited the highest activities. Using different chromatographic techniques, four compounds were isolated and identified as rutin, gallic acid, nictoflorin, and tulipanin. In conclusion: The ethyl acetate fraction of the rind of Punica granatum var. nana may provide a potential therapeutic approach for hyperglycemia.

Functional targeting of the TGF-ßR1 kinase domain and downstream signaling: A role for the galloyl moiety of green tea-derived catechins in ES-2 ovarian clear cell carcinoma.

The galloyl moiety is a specific structural feature which dictates, in part, the chemopreventive properties of diet-derived catechins. In ovarian cancer cells, galloylated catechins were recently demonstrated to target the transforming growth factor (TGF)-ß-mediated control of the epithelial-mesenchymal transition process. The specific impact of the galloyl moiety on such signaling, however, remains poorly understood. Here, we questioned whether the sole galloyl moiety interacted with TGF-ß-receptors to alter signal transduction and chemotactic migratory response in an ES-2 serous carcinoma-derived ovarian cancer cell model. In line with the LogP and LogS values of the tested molecules, we found that TGF-ß-induced Smad-3 phosphorylation and cell migration were optimally inhibited, provided that the lateral aliphatic chain of the galloyl moiety reached 8-10 carbons. Functional inhibition of the TGF-ß receptor (TGF-ßR1) kinase activity was supported by surface plasmon resonance assays showing direct physical interaction between TGF-ßR1 and the galloyl moiety. In silico molecular docking analysis predicted a model where galloylated catechins may bind TGF-ßR1 within its adenosine triphosphate binding cleft in a site analogous to that of Galunisertib, a selective adenosine triphosphate-mimetic competitive inhibitor of TGF-ßR1. In conclusion, our data suggest that the galloyl moiety of the diet-derived catechins provides specificity of action to galloylated catechins by positioning them within the kinase domain of the TGF-ßR1 in order to antagonize TGF-ß-mediated signaling that is required for ovarian cancer cell invasion and metastasis.

Elaeocarpus floribundus Bl. seeds as a new source of bioactive compounds with promising antioxidant and antimicrobial properties.

The fruits of Elaeocarpus floribundus Bl. (Elaeocarpaceae) are edible and are normally prescribed for treatment of diseases. The medicinal uses of the fruit create considerable quantities of seeds as wastes. In an attempt to valorise this biomass, we studied the antimicrobial and antioxidant activities of the ethanolic seed extract. The extract inhibited the growth of the tested pathogens and was also a very strong scavenger of DPPH free radicals. Consequently, the extract was phytochemically investigated and this study reports the initial isolation of five phenolic compounds from this source. The structures of the isolated compounds were elucidated by spectra analyses including HR-ESI-MS, 1D and 2D NMR experiments. The isolated compounds exhibited a wide range of antimicrobial activities against the tested pathogens. Gallic acid (4) showed the most activity against Bacillus subtilis with a minimum inhibitory concentration (MIC) value of 30 µg/mL, while the MIC values of the antimicrobial standards range between 10 and 35 µg/mL. Compound 4, crude ethanolic extract, and the ethyl acetate fraction were more a potent free radical scavenger of DPPH compared to ascorbic acid. Hence, the seeds of E. floribundus could be considered as a new source of bioactive compounds for pharmaceutical and food-related industries.

Subcritical water extraction of bioactive compounds from Orostachys japonicus A. Berger (Crassulaceae).

Subcritical-water extraction is an ecofriendly method for extracting antioxidant compounds only using water. The Subcritical-water extraction was employed for the extraction of bioactive compounds from Orostachys japonicus known as rock pine by investigating the use of various temperatures (110-260 °C) and extraction times (5-20 min). The Subcritical-water extraction condition at 220 °C for 15 min; the total phenolics content (39.9 ± 4.1 mg/g), flavonoids content (11.4 ± 0.6 mg/g), and antioxidant activities (90.3 ± 2.2%, 96.0 ± 2.9%, and 662.4 ± 17.2 mg/g) of Subcritical-water extract were higher under this condition than for extraction with either methanol or ethanol. Triterpene saponins were observed only in subcritical-water extraction condition at 220 °C for 15 min. Further, some of its phenolic constituents; gallic acid, quercetin, and kaempferol were quantified by high performance liquid chromatography. Subcritical-water extraction is an effective method for extracting valuable bioactive compounds from Orostachys japonicus.

Analysis of polyphenolic metabolites from in vitro gastrointestinal digested soft fruit extracts identify malvidin-3-glucoside as an inhibitor of PTP1B.

Protein-tyrosine phosphatase 1B (PTP1B, EC 3.1.3.48) is an important regulator of insulin signalling. Herein, we employed experimental and computational biology techniques to investigate the inhibitory properties of phenolics, identified from four in vitro gastrointestinal digested (IVGD) soft fruits, on PTP1B. Analysis by LC-MS/MS identified specific phenolics that inhibited PTP1B in vitro. Enzyme kinetics identified the mode of inhibition, while dynamics, stability and binding mechanisms of PTP1B-ligand complex were investigated through molecular modelling, docking, molecular dynamics (MD) simulations, and MM/PBSA binding free energy estimation. IVGD extracts and specific phenolics identified from the four soft fruits inhibited PTP1B (P < 0.0001) activity. Among the phenolics tested, the greatest inhibition was shown by malvidin-3-glucoside (P < 0.0001) and gallic acid (P < 0.0001). Malvidin-3-glucoside (Ki = 3.8 µg/mL) was a competitive inhibitor and gallic acid (Ki = 33.3 µg/mL) a non-competitive inhibitor of PTP1B. Malvidin-3-glucoside exhibited better binding energy than gallic acid and the synthetic inhibitor Dephostatin (-7.38 > -6.37 > -5.62 kcal/mol) respectively. Principal component analysis demonstrated malvidin-3-glucoside PTP1B-complex occupies more conformational space where critical WPD-loop displayed a higher degree of motion. MM/PBSA binding free energy for malvidin-3-glucoside to PTP1B was found to be higher than other complexes mediated by Van der Waals energy rather than electrostatic interaction for the other two inhibitors (-80.32 ± 1.25 > -40.64 ± 1.43 > -21.63 ± 1.73 kcal/mol) respectively. Altogether, we have established novel insights into the specific binding of dietary phenolics and have identified malvidin-3-glucoside as an PTP1B inhibitor, which may be further industrially developed for the treatment of type-2 diabetes.

Protection of the Geum japonicum Thunb. var. chinense extracts against oxygen-glucose deprivation and re-oxygenation induced astrocytes injury via BDNF/PI3K/Akt/CREB pathway.

Geum japonicum Tunb. var. chinense (GJ) is a traditional Chinese medicine usually used for the alleviation of dizziness and headache. Previous studies have reported that the GJ extracts could alleviate cerebral I/R injury by reducing apoptosis in vivo. To further elucidate the positive role and underlying mechanism of the GJ extracts in cerebral I/R injury, the current study investigated the effects of the GJ extracts on oxygen-glucose deprivation and re-oxygenation (OGD/R)-induced astrocytes injury in light of BDNF/PI3K/Akt/CREB signaling pathway with seropharmacological method. In the present study, the LC-MS profiling of the GJ extracts, obtain by reflux extraction, led to the identification of three possible active components were 5-desgalloylstachyurin, tellimagrandin II (TG II) and 3,4,5-Trihydroxybenzaldehyde (THBA). Drug-containing serum was collected from rats given different doses of the GJ extracts (0, 1.75 g/kg, 7 g/kg). Data indicated that the GJ extracts could increase the cell viability and decrease apoptosis and the expression of glial fibrillary acidic protein (GFAP) in OGD/R-induced astrocytes. In addition, the detection of apoptosis-related factors showed that the GJ extracts could obviously increase the expression of Bcl-2 and reduce the expression of Bax, Caspase-3 and cleaved-Caspase-3. Furthermore, the GJ extracts markedly increased the expression of BDNF, TrkB, PI3K, p-Akt and p-CREB. All these effects of the GJ extracts could be significantly reversed by LY294002, an inhibitor of PI3K. These data indicated that the GJ extracts could protect astrocytes against OGD/R-induced injury by inhibiting astrocytes reactivity and apoptosis, owing to the activation of the BDNF/PI3K/Akt/CREB pathway. The results support the application of the GJ extracts in the treatment of ischemic stroke and other ischemic encephalopathy.

Simultaneous extraction and separation of bioactive compounds from apple pomace using pressurized liquids coupled on-line with solid-phase extraction.

The objective of this work was the development of an on-line extraction/fractionation method based on the coupling of pressurized liquid extraction and solid-phase extraction for the separation of phenolic compounds from apple pomace. Several variables of the process were evaluated, including the amount of water of the first stage (0-120 mL), temperature (60-80 °C), solid-phase extraction adsorbent (Sepra, Isolute, Strata X and Oasis) and activation/elution solvent (methanol and ethanol). The best results were observed with the adsorbent Sepra. The temperature had a small effect on recovery, but significant differences were observed for phlorizin and a quercetin derivative. Results indicate that ethanol can be used to replace methanol as an activation, extraction/elution solvent. While using mostly green solvents (water, ethanol, and a small amount of methanol that could be reused), the developed method produced higher or similar yields of acids (2.85 ± 0.19 mg/g) and flavonoids (0.97 ± 0.11 mg/g) than conventional methods.

Potential Application of Hippophae Rhamnoides in Wheat Bread Production.

Sea buckthorn (Hippophae rhamnoides) berries are well known for their content in bioactive compounds, high acidity, bright yellow color, pleasant taste and odor, thus their addition in a basic food such as bread could be an opportunity for modern food producers. The aim of the present research was to investigate the characteristics and the effects of the berry' flour added in wheat bread (in concentration of 1%, 3% and 5%) on sensory, physicochemical and antioxidant properties, and also bread shelf life. Berry flour contained total polyphenols-1467 mg gallic acid equivalents (GAE)/100 g, of which flavonoids-555 mg GAE/100 g, cinnamic acids-425 mg caffeic acid equivalents (CAE)/100 g, flavonols-668 mg quercetin equivalents (QE)/100 g. The main identified phenolics were catechin, hyperoside, chlorogenic acid, cis- and trans-resveratrol, ferulic and protocatechuic acids, procyanidins B1 and B2, epicatechin, gallic acid, quercetin, p- and m-hydroxybenzoic acids. The antioxidant activity was 7.64 mmol TE/100 g, and carotenoids content 34.93 ± 1.3 mg/100 g. The addition of berry flour increased the antioxidant activity of bread and the shelf life up to 120 h by inhibiting the development of rope spoilage. The obtained results recommend the addition of 1% Hippophae rhamnoides berry flour in wheat bread, in order to obtain a product enriched in health-promoting biomolecules, with better sensorial and antioxidant properties and longer shelf life.