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The present study was designed to evaluate the chemical extraction, chemical composition, and antioxidant and antibacterial properties of the total flavonoids in Willow Buds (TFW). We investigated the optimal extraction of TFW using response surface methodology (RSM). Chemical compounds were analyzed using Q-Orbitrap LC-MS/MS. The DPPH radical scavenging capacity, hydroxy radical inhibitory ability, and superoxide anion radical inhibitory ability were explored to determine the antioxidant properties of flavonoid extractions. The antibacterial effect was assessed via minimal inhibitory concentration. The results demonstrated that the optimal extraction conditions were an ethanol concentration of 50%, a time of 35 min, and a liquid/material ratio of 70:1 mL/g. Under these conditions, the yield of TFW was 7.57%. Eight flavonoids, a phenolic glycoside, and an alkaloid were enriched in the Willow Buds. The TFW exhibited significant antioxidant activity, with IC 50 values of 0.18-0.24 mg/mL and antimicrobial activity against Escherichia coli , Salmonella enterica , Staphylococcus aureus , and Streptococcus pneumoniae . TFW may be explored as potential and natural compounds in food and pharmacological applications.

Cajaninstilbene acid (CSA), an active compound separated from pigeon pea leaves, possesses the highly efficient antioxidant activities. Transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf2) is an important regulator of cellular oxidative stress. This study examined the role of Nrf2 in CSA-mediated antioxidant effects on human hepatocarcinoma (HepG2) cell line. The generation of reactive oxygen species (ROS) upon H2O2 and CSA treatment was lower than that of H2O2 alone. CSA activated Nrf2 as evaluated by Western blotting. A luciferase reporter assay also demonstrated that CSA-activated signaling resulted in the increased transcriptional activity of Nrf2 through binding to the antioxidant response element (ARE) enhancer sequence. Our study indicated that treatment of HepG2 cells with CSA induces Nrf2-dependent ARE activity and gene expression of heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase 1 (NQO1), and glutamate-cysteine ligase modifier subunits by activation of PI3K/AKT, ERK and JNK signaling pathways. Inhibition of Nrf2 by siRNA reduced CSA-induced upregulation of these Nrf2-related enzymes. These results suggest that the Nrf2/ARE pathway plays an important role in the regulation of CSA-mediated antioxidant effects in HepG2 cells., (Crown Copyright © 2013. Published by Elsevier Ireland Ltd. All rights reserved.)

Dihydroquercetin (DHQ) is a well-known antioxidant agent. In the present investigation, we reported for the first time that DHQ stimulates the expression of phase II detoxifying enzymes through the Nrf2-dependent signaling pathway. The IC50 values of DHQ for reduction of 2,2-diphenyl-1-picrylhydrazol (DPPH), reducing power assay, lipid peroxidation assay, and xanthine oxidase inhibition were 5.96, 4.31, 2.03, and 13.24 μM, respectively. DHQ possessed considerable protective activity from oxidative DNA damage. A luciferase reporter assay also demonstrated that DHQ-activated signaling resulted in the increased transcriptional activity of Nrf2 through binding to the ARE (antioxidant response element) enhancer sequence. Furthermore, Western blotting and luciferase assay revealed DHQ activated ERK1/2, Akt, and JNK signaling pathways, subsequently leading to Nrf2 nuclear translocation. DHQ upregulated the Nrf2-related antioxidant genes heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase-1 (NQO1), and glutamate-cysteine ligase modifier subunits. Inhibition of Nrf2 by siRNA reduced DHQ-induced upregulation of these antioxidant genes.

In this study, novel endophytic fungi producing cajaninstilbene acid (CSA) from pigeon pea [ Cajanus cajan (L.) Millsp.] were investigated and screened. CSA has prominent pharmacological activities. A total of 110 endophytic fungi isolates were grouped into 8 genera on the basis of morphological characteristics, and CSA-producing fungi were screened by liquid chromatography-tandem mass spectrometry (LC-MS/MS). According to ITS-rDNA sequences analysis, the CSA-producing fungi were identified as Fusarium solani (ERP-07), Fusarium oxysporum (ERP-10), and Fusarium proliferatum (ERP-13), respectively. The amount of CSA produced by the ERP-13 reached 504.8 ± 20.1 μg/L or 100.5 ± 9.4 μg/g dry weight of mycelium. In a DPPH radical-scavenging assay, when the concentration of fungal CSA was 500 μg/mL, inhibition percentage could reach 80%, which was almost the same as that of standard CSA. This study first reported the natural antioxidant CSA from endophytic fungi F. solani and F. proliferatum isolated from pigeon pea.

In this study, the aqueous and ethanolic extracts (leaves, stems and fruits) from Morus alba L., a traditional Chinese medicine, were evaluated for their antioxidant and antimicrobial properties. Ethanolic extracts showed higher contents of both total phenolics and flavonoids than aqueous extracts. The total phenolic content was in the order of: leaf extracts > fruit extracts > stem extracts, whereas the total flavonoids was: leaf extracts > stem extracts > fruit extracts. Using DPPH assays, the concentrations providing 50% inhibition (IC(50)) values of aqueous extracts from leaves, stems and fruits were 7.11 ± 1.45 mg/ml, 86.78 ± 3.21 mg/ml and 14.38 ± 2.83 mg/ml, respectively, whereas the IC(50) values of ethanolic extracts were 3.11 ± 0.86 mg/ml, 14.62 ± 2.45 mg/ml and 12.42 ± 2.76 mg/ml, respectively. In sum, the antioxidant activities of ethanolic extracts from M. alba L. were stronger than the aqueous extracts, and in the order of: leaf extracts > fruit extracts > stem extracts. The ethanolic extracts exhibited moderate antimicrobial activities, whereas the aqueous extracts showed poor antimicrobial properties in our test system. This study validated the medicinal potential of M. alba L.

The antioxidant properties, DNA damage protective activities, and xanthine oxidase (XOD) inhibitory effect of cajaninstilbene acid (CSA) derived from pigeon pea leaves were studied in the present work. Compared with resveratrol, CSA showed stronger antioxidant properties, DNA damage protective activity, and XOD inhibition activity. The IC(50) values of CSA for superoxide radical scavenging, hydroxyl radical scavenging, nitric oxide scavenging, reducing power, lipid peroxidation, and XOD inhibition were 19.03, 6.36, 39.65, 20.41, 20.58, and 3.62 μM, respectively. CSA possessed good protective activity from oxidative DNA damage. Furthermore, molecular docking indicated that CSA was more potent than resveratrol or allopurinol to interact with the active site of XOD (calculated free binding energy: -229.71 kcal mol(-1)). On the basis of the results, we conclude that CSA represents a valuable natural antioxidant source and may potentially be applicable in health food industry.

The antioxidant capacity and xanthine oxidase inhibitory effects of extracts and main polyphenolic compounds of Geranium sibiricum were studied in the present work. The antioxidant capacity was evaluated by ferric reducing antioxidant power, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, superoxide radical scavenging, nitric oxide scavenging, beta-carotene-linoleic acid bleaching, and reducing power assays. Among the extracts and four fractions, the ethyl acetate fraction showed the highest phenolic content (425.36 +/- 9.70 mg of gallic acid equivalent/g extracts) and the best antioxidant activity. The IC(50) values of the ethyl acetate fraction were 0.93, 3.32, 2.06, 2.66, and 1.64 microg/mL in the DPPH radical scavenging, superoxide radical scavenging, nitric oxide scavenging, beta-carotene-linoleic acid bleaching, and reducing power assays, respectively. Of the polyphenolic compounds separated from the ethyl acetate fraction, geraniin showed a higher activity than corilagin and gallic acid. The IC(50) values ranged from 0.87 to 2.53 microM, which were even lower than the positive control (except for allopurinol). All test samples except for the petroleum ether fraction showed xanthine oxidase inhibitory effects. We conclude that G. sibiricum represents a valuable natural antioxidant source and is potentially applicable in the healthy food industry.

Phellodendron amurense Rupr. bark extracts were examined for antioxidant activity, antimicrobial activity and antiviral activity on herpes simplex virus type 1 (HSV-1). Ethanol extract showed higher content of both total phenolic and flavonoid than aqueous extract. In DPPH (2, 2-diphenyl-1-picrylhydrazyl) assay, the concentration providing 50% inhibition (IC(50)) values were 6.73 +/- 0.87 mg/ml and 4.26 +/- 0.59 mg/ml for aqueous and ethanol extracts respectively. Ethanol extract had a much higher antimicrobial activity than aqueous extract. Furthermore, the antiviral activity of the ethanol extract on HSV-1 was also tested. The maximum noncytotoxic concentration was 44.12 microg/ml for RC-37 cells. Plaque formation was inhibited by 74+/-6% when HSV-1 was pretreated with the extract prior to adsorption, whereas pretreatment of the cells with the extract, added during adsorption or after the adsorption only exhibited above 10% antiviral effect. This study has to some extent validated the medicinal potential of P. amurense bark.

Artemisia absinthium L. is a highly medicinal plant with a broad range of biomedical applications. A. absinthium callus cultures were established in response to bio-fabricated single NPs (Ag and Cu) or a combination of both NPs (Ag and Cu) in different ratios (1:2, 2:1, 1:3, and 3:1) along with thidiazuron (TDZ) (4 mg/L) to elicit the biomass accumulation, production of non-enzymatic compounds, antioxidative enzymes, and antioxidant activity. Silver and copper nanoparticles (Ag and Cu NPs) were synthesized using the leaves of Moringa oleifera as reducing and capping agent and further characterized through UV-Visible spectroscopy and SEM. The 30 µg/L suspension of Ag and Cu NPs (1:2, 2:1) and 4 mg/L TDZ showed 100% biomass accumulation as compared to control (86%). TDZ in combination with Ag NPs enhanced biomass in the log phases of growth kinetics. The Cu NPs alone enhanced the superoxide dismutase activity (0.56 nM/min/mg FW) and peroxidase activity (0.31 nM/min/mg FW) in callus cultures. However, the combination of Ag and Cu NPs with TDZ induced significant total phenolic (7.31 µg/g DW) and flavonoid contents (9.27 µg/g DW). Furthermore, the antioxidant activity was highest (86%) in the Ag and Cu NPs (3:1) augmented media. The present study provides the first evidence of bio-fabricated single NPs (Ag and Cu) or a combination of both NPs (Ag and Cu) in different ratios (1:2, 2:1, 1:3, and 3:1) along with TDZ (4 mg/L) on the development of callus culture, production of endogenous enzymes, non-enzymatic components, and further antioxidant activity in callus cultures of A. absinthium.

The objective of this research was twofold: first, to optimize the extraction process of Lonicera japonica polyphenols using a response surface methodology, and second, to study the antioxidant activity and tyrosinase inhibitory capacity of the polyphenols of different purities. High‐speed shearing homogenization extraction was used to extract the polyphenols from L. japonica. The antioxidant activity and the effect of polyphenols on tyrosinase activity were studied using free radical scavenging assay and the tyrosinase method, respectively. The optimal extraction conditions with an extraction yield of 6.96% for polyphenols were determined as follows: ethanol volume fraction 57%, shearing time 3.30 min, and solid–liquid ratio 1:58. Lonicera japonica polyphenols exhibited potent scavenging activity on 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and 2, 2'‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS), and inhibitory capacity on tyrosinase. The results suggested that L. japonica polyphenols could be explored as a natural antioxidant and tyrosinase inhibitor. [ABSTRACT FROM AUTHOR]

Fig (Ficus carica L.) leaves are produced each year and often disposed, resulting in a waste of resources. Fig waste leaves are rich in flavonoids, which have strong antioxidant activity; however, the variety and chemical structure of antioxidants in fig leaves have not been reported in detail. To take full advantage of fig waste leaves, antioxidant capacity of different extracts (petroleum ether, ethyl acetate, and water) was evaluated by 1, 1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic) acid (ABTS), and ferric-ion-reducing antioxidant power (FRAP) methods. The results showed that flavonoids in ethyl acetate extraction had the highest content (83.92 ± 0.01 mg/g), maximum DPPH scavenging activity (IC50 0.54 mg/mL), highest ABTS scavenging rate (80.28%), and FRAP (3.46 mmol/g). Furthermore, an HPLC-QTOF-MS/MS-DPPH method was developed to identify 11 flavonoids in fig waste leaves. This rapid and efficient method can not only be used for screening the antioxidant components in fig waste leaves, but also can be combined with mass spectrometry to identify the compounds with antioxidant capacity. There are three flavonoids with significant antioxidant capacity, which are 3-O-(rhamnopyranosyl-glucopyranosyl)-7-O-(glucopyranosyl)-quercetin, isoschaftoside, and rutin. The results confirmed that fig waste leaves contain a variety of antioxidant components, which contributed to increase the value of fig waste leaves as antioxidants. [ABSTRACT FROM AUTHOR]

Abstract: This study evaluated the antioxidant activity of the main active compound of an endophytic fungus MD-R-1 from pigeon pea. This fungus was identified as Aspergillus fumigates by ITS sequence. The antioxidant capacity was evaluated by DPPH, hydroxyl radical scavenging, reducing power, lipid peroxidation and XOD inhibitory effect assays. Among the four fractions, the EtOAc fraction showed the best antioxidant activity (IC50 values were 16.38, 35.20, 19.69, 22.64 and 193.24μg/mL, respectively) and the highest phenolic and flavonoid content. Furthermore, it possessed good protective activity from oxidative DNA damage. The activities of superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) in HepG2 cells treated with MD-R-1 EtOAc extracts were also determined. The results showed that EtOAc extracts significantly increased the activities of SOD, CAT and GR in HepG2 cells in a dose-dependent manner. The LC–MS/MS was used to analyze EtOAc fraction and the result pointed to a notable antioxidant compound, luteolin. The present investigation suggests that MD-R-1 represents a valuable natural antioxidant source and may be applicable in health food industry potentially. [Copyright &y& Elsevier]

Abstract: A fungal endophyte MD89 with obvious antioxidant activities was isolated from pigeon pea and identified as Chaetomium globosum by ITS sequence. Different fractions from MD89 culture were compared and evaluated by total phenol (TP) content, total flavonoid (TFL) content, DPPH radical scavenging, reducing power and lipid peroxidation assays, respectively. Results showed that EtOAc extracts had high content of TP and TFL, and strong antioxidant activities (IC50 value was 6.87, 15.19, 16.78μg/mL, respectively). Furthermore, the EtOAc extracts were analyzed by LC–MS/MS and a good antioxidant compound, apigenin, was found. The activities of superoxide dismutase (SOD) and glutathione reductase (GR) in HepG2 cells treated with MD89 EtOAc extracts were also determined. It is revealed that EtOAc extracts increased the activities of SOD and GR in HepG2 cells significantly in a dose-dependent manner. On the basis of those results, we conclude that MD89 represents a valuable natural antioxidant source and may be applicable in health food industry potentially. [Copyright &y& Elsevier]

Abstract: Applicability of solvent-free microwave extraction (SFME) for extraction of the fruits of Schisandra chinensis essential oil was examined; the composition and antioxidant activities and antibacterial activities of the essential oil were assessed in vitro. An orthogonal experiment (L9 (3)4) was applied to optimize the extraction process. The optimum conditions were: extraction time, 45 min; microwave power, 800 W; diameter of powder particles, 0.25 mm; and proportion of water pretreatment, 30%. Under these conditions, the extraction yield was 1.75%. Thirty-five compounds, representing 91.12% of the oil, were identified, of which the major ones, ylangene (50.11%), β-himachalene (10.76%)，α-bergamotene (9.52%) and β-Chamigrene (5.41%), accounted for of 75.80% the oil. Antioxidant activity, IC50 value of the essential oil was determined as 3.87 mg/mL by DPPH assay, and the inhibition values of the essential oil at 1.8 mg/mL was 41.88% by β-Carotene–linoleic acid bleaching assay. The essential oil was screened for antibacterial activity against both Gram positive (Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis) and Gram negative (Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris) bacteria. The essential oil showed antibacterial effect against all the gram (+) bacteria and gram (−) bacteria tested. These results show that S. chinensis essential oil could be considered as a natural alternative to food antioxidants and preservatives. [Copyright &y& Elsevier]

Abstract: An enzyme incubation–water extraction (EI–WE) method was developed and optimised for the extraction of the natural antioxidant taxifolin and of the total flavonoids from wood sawdust of Larix gmelini (Rupr.) Rupr. A factorial design and a central composite design approach were used for method optimisation. Optimal conditions were 0.5mg/ml cellulase and 0.5mg/ml pectinase, a pH of 5.0, a temperature of 32°C and 18h incubation time. The flavonoids and taxifolin were extracted in hot water at 50°C for 30min, with a solid to liquid ratio of 1:20. Under optimised conditions, the yields of taxifolin and total flavonoids increased from 1.06±0.08 to 1.35±0.04mg/g and 4.13±0.17 to 4.96±0.29mg/g, respectively. DPPH and BHT assays revealed that the EI–WE samples had 1.8- and 1.68-fold higher antioxidant activities than the controls. SEM results revealed the structural disruption of wood sawdust with enzyme incubation. [ABSTRACT FROM AUTHOR]