Enterolactone restricts the proliferation of the LNCaP human prostate cancer cell line in vitro

Ecological data suggest a long‐term diet high in plant material rich in biologically active compounds, such as the lignans, can significantly influence the development of prostate cancer over the lifetime of an individual. The capacity of a pure mammalian lignan, enterolactone (ENL), to influence the proliferation of the LNCaP human prostate cancer cell line was investigated as a function of cell density, metabolic activity, expression and secretion of prostate specific antigen (PSA), cell cycle profile, and the expression of genes involved in development and progression of prostate cancer. Treatment with a subcytotoxic concentration of ENL (60 μM for 72 h) was found to reduce: cell density (57.5%, SD 7.23, p < 0.001), metabolic activity (55%, SD 0.03, p < 0.001), secretion of PSA (48.50% SD 4.74, p = 0.05) and induce apoptosis (8.33‐fold SD 0.04, p = 0.001) compared to untreated cells. Cotreatment with 10 μM etoposide was found to increase apoptosis by 50.17% (SD 0.02, p < 0.001). Additionally, several key genes (e. g. MCMs, survivin and CDKs) were beneficially regulated by ENL treatment (p < 0.05). The data suggest that the antiproliferative activity of ENL is a consequence of altered expression of cell cycle associated genes and provides novel molecular evidence for the antiproliferative properties of a pure lignan in prostate cancer.     

Heat treatment enhances the NO-suppressing and peroxynitrite-intercepting activities of kumquat (Fortunella margarita Swingle) peel

In Taiwan, folk remedies containing dried kumquats (Fortunella margarita Swingle) are used to cure inflammatory respiratory disorders. The induction of inducible nitric oxide (NO) synthase in inflammatory cells and increased airway production of NO and peroxynitrite, its derivative, are key events in such disorders. Although heat is known to affect the antioxidant activity of citrus peels, the effects of dehydration and heating on NO suppression and on the interception of peroxynitrite are unclear. We determined the NO-suppressing activities of freeze-dried, oven-dried, and heat-treated kumquat extracts by measuring their inhibition of NO production in lipopolysaccharide-activated RAW 264.7 macrophages. Furthermore, we evaluated the attenuation of peroxynitrite-mediated nitrotyrosine formation in albumin. Heating, but not oven drying, enhanced the ability of kumquat peels to suppress NO and intercept peroxynitrite, as compared with freeze drying. However, heat treatment and oven drying of kumquat flesh attenuated these activities; these effects were at least partially attributed to heat-susceptible ascorbate.     

Effect of heat treatments on the essential oils of kumquat (Fortunella margarita Swingle)

Kumquats (Fortunella margarita Swingle) cultivated in Taiwan are eaten raw or made into candied fruit or fruit tea. For the experiments described in this paper, essential oils were obtained from kumquat peels or whole fruit by cold pressing, steam distillation or heating in water at 90 °C for 15 min followed by steam distillation. The volatile components contained in the essential oils were identified by direct injection (DI) or headspace-solid phase microextraction (HS-SPME) coupled with gas chromatography (GC). A total of 43 compounds were identified, of which 37 were verified by DI/GC and 31 by HS-SPME/GC. Hot water heating increased the yields of essential oils from both peels and whole fruit. The principal constituents of the oils were similar except for the minor compounds, including linalool, terpinen-4-ol and α-terpineol, the levels of which increased after steam distillation. The whole fruit also contained higher levels of terpene alcohols.     

Radical scavenging and antibacterial properties of the extracts from different Thymus pulegioides L. chemotypes

Radical scavenging and antibacterial properties of large thyme extracts isolated from five chemotypes of Thymus pulegioides L. growing wild in Lithuania were studied. The chemotypes were defined according to the main essential oil components: linalool (L), geranial/geraniol/neral (G/G/N), thymol (T), carvacrol/γ-terpinene/p-cymene (C/γT/pC) and thymol/carvacrol/γ-terpinene/p-cymene (T/C/γT/pC). The contents of phenolic compounds, flavonoids and flavonols were determined. It was found that the extracts of phenolic chemotypes containing remarkable concentrations of thymol and/or carvacrol were stronger DPPH and ABTS free radical scavengers in the model systems. The antibacterial activity of the extracts depended on the plant chemotype, extract preparation, solvent used and finally the sensitivity of bacteria. Bacillus cereus, Micrococcus luteus, Staphylococcus epidermidis and Staphylococcus aureus were the most sensitive to the all extracts applied, whereas Escherichia coli, Salmonella typhimurium and Enterobacter aerogenes remained resistant.     

Kumquat (Fortunella japonica Swingle) juice: Flavonoid distribution and antioxidant properties

The flavonoids composition of kumquat (Fortunella japonica) crude juice, obtained from uripe and ripe fruits, were investigated by reverse-phase LC-DAD-ESI-ITMS analysis. Thirteen compounds (C- and O-glycosyl flavonoids) were identified. Acacetin 3,6-di-C-glucoside (1), vicenin-2 (2), lucenin-2 4′-methyl ether (3) narirutin 4′-O-glucoside (4) and apigenin 8-C-neohesperidoside (5) were identified for the first time in kumquat juice. Furthermore, we report on the remarkable antioxidant properties of crude juice, of selected flavonoids fractions and of the main component, phloretin 3′,5′-di-C-glucoside (6).     

Antioxidant capacities of Gundelia tournefortii L. extracts and inhibition on glutathione-S-transferase activity

Gundelia tournefortii L. is an important food source and a well-known medicinal plant in Eastern Anatolia. Therapeutic effects of medicinal plants are known to be closely related to their antioxidant capacities. Antioxidant activities of G. tournefortii, both for the aerial parts and seeds, were investigated by using both 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and lipid peroxidation inhibition methods. The seeds were found to have higher antioxidant potential than the aerial, with IC₅₀ values of 0.073 mg/mL for DPPH scavenging and 0.146 mg/mL for lipid peroxidation inhibition capacities. In addition, total phenolic contents of the Gundelia tournefortii L. extracts, especially the seed extracts correlates to its high antioxidant activity with 105.1 ± 8.7 μg gallic acid equivalents (GAEs) per mg of seed extract. Plant extracts with high phenolics content are known to have important effects on various enzymes, as well as glutathione-S-transferases, which are important detoxification enzymes in phase II systems with an important role in developing multi-drug resistance to chemotherapy in tumour cells. Consequently, the effects of G. tournefortii extracts on crude cytosolic glutathione-S-transferase was also studied and the seed extracts have shown effective inhibition of cytosolic GST activity, with an IC₅₀ of 97.51 μg/mL.     

Apoptosis-mediated proliferation inhibition of human colon cancer cells by volatile principles of Citrus aurantifolia

Fruits of Citrus aurantifolia were subjected to hydro-distillation using Clevenger type apparatus to obtain volatile oil. Chemical composition of volatile oil was analysed by GC–MS. Twenty-two compounds representing more than 89.5% of the volatile oil were identified. d-limonene (30.13%) and d-dihydrocarvone (30.47%) were found to be the major compounds in the lime volatile oil. This oil showed 78% inhibition of human colon cancer cells (SW-480) with 100 μg/ml concentration at 48 h. Lime volatile oil showed DNA fragmentation and induction of caspase-3 up to 1.8 and 2- folds after 24 h and 48 h, respectively, which may be due to the involvement of apoptosis. Analysis of apoptosis-related protein expression further confirmed apoptosis induction by lime volatile oil. The above results suggested that lime volatile oil has potential benefits in colon cancer prevention. This is the first report, showing the possible mechanism of antiproliferative effect of lime volatile oil for the prevention of colon cancer in cell culture models.     

Major anthocyanins and antioxidant activity of Nasturtium flowers (Tropaeolum majus)

Major anthocyanins, ascorbic acid content, total phenolic content, and the radical scavenging activity against ABTS and DPPH radicals in petals of orange Nasturtium flowers (Tropaeolum majus), were investigated. Anthocyanin (ACN) content in the petals was 72 mg/100 g FW and pelargonidin 3-sophoroside represented 91% of the total ACN content. The ascorbic acid content was 71.5 mg/100 g and the total phenolic content as determined by the Folin–Ciocalteau method was 406 mg GAE/100 g FW. The radical scavenging activities against ABTS and DPPH radicals were 458 and 91.87 μm trolox eq/g FW, respectively. The excellent free radical scavenging activities along with high phenolic and ascorbic acid content of Nasturtium flowers suggest that they could be source of natural pigments and antioxidants for applications in functional foods.     

Radical scavenging and reducing properties of extracts of cashew shoots (Anacardium occidentale)

The total phenolic content and antioxidant activities of methanol, hexane and ethyl acetate extracts of the shoots of Anacardium occidentale were measured. Total phenolic content was assessed by the Folin–Ciocalteau assay whereas antioxidant activities were assessed by measuring the ability of the extracts to scavenge the ABTS^·+ and DPPH^· radicals, superoxide anion radicals and nitric oxide radicals as well as their ability to reduce ferric ions. Results indicated that the methanol extract of A. occidentale was the most potent reducing agent and radical-scavengers compared to the other two extracts. The ethyl acetate extract exhibited some antioxidant activities whereas the hexane extract is the least reactive. The order of the antioxidant potency of the plant extract is methanol > ethyl acetate > hexane. The methanol extract contained more than 7 fold of total phenolic content compared to the hexane and ethyl acetate extracts indicating the likely possibility that the observed antioxidant activities were partly contributed by the phenolics. The results suggest that the shoots of A. occidentale are a source of natural antioxidants.     

Antioxidative activity of Mungoong,an extract paste,from the cephalothorax of white shrimp (Litopenaeus vannamei)

The antioxidative activity of Mungoong, an extract paste, from the cephalothorax of white shrimp (Litopenaeus vannamei) was studied. Extraction media were shown to affect the antioxidative activity and properties of resulting extracts from Mungoong. Distilled water exhibited the highest efficacy in extracting the antioxidants from Mungoong, as evidenced by the highest ABTS (2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) and DPPH (2,2-diphenyl-1-picryl hydrazyl) radical scavenging activity as well as ferric reducing activity power (FRAP), compared with distilled water/ethanol mixture (1:1, 1:2 and 2:1) and ethanol. UV-absorbances at both 280 and 295 nm (A₂₈₀, A₂₉₅), browning intensity (A₄₂₀) and fluorescence intensity were also highest in the extract using distilled water. ABTS and DPPH radical scavenging activity and FRAP of water extract, increased linearly with increasing concentration. Good correlation between ABTS and DPPH radical scavenging activity; DPPH radical scavenging activity and FRAP; ABTS radical scavenging activity and FRAP were observed, suggesting that antioxidants in the extract, possessed the capability of scavenging the radicals together with reducing power. Antioxidants in the water extract from Mungoong showed high stability over the wide pH ranges (2–11) and temperature up to 100 °C, in which the activity of more than 80% remained. MALDI-TOF analysis revealed that water extract contained the peptides having the mass ranges of m/z 400–1000 and 4000–7000.     

Antioxidant and Free Radical Scavenging Activities of Indian Acacias: Acacia Leucophloea (Roxb.) Willd., Acacia Ferruginea Dc., Acacia Dealbata Link. and Acacia Pennata (L.) Willd

The acetone and methanol extracts of barks of A. leucophloea, A. ferruginea, A. dealbata, and A. pennata were evaluated for their total phenolics and flavonoid contents and antioxidative activities. Acetone extracts of barks exhibited higher contents of total phenolics and flavonoids. Further, the acetone extracts showed higher activity in DPPH^?, ABTS^?+, and OH^?; FRAP; metal chelation; phosphomolybdenum reduction; and peroxidation inhibition. In conclusion, the results implied that the barks could be considered as health supplements and nutraceuticals/functional foods.     

Characterization of the phytochemicals and antioxidant properties of extracts from Teaw (Cratoxylum formosum Dyer)

The leaves of the Thai vegetable, Teaw (Cratoxylum formosum Dyer) were extracted with ethanol to provide an extract that had antioxidant properties. The composition of the extract was studied by high-performance liquid chromatography with a diode array detector, and by electrospray ionization mass spectrometry. The main antioxidant component (peak 1) was chlorogenic acid, which was present at 60% of the extract. Three minor components were present at 7%, 3% and 2%, and other components that were present at lower concentrations were also observed. Treatment of the Teaw extract with 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) caused a similar reduction in peak area of 55.2–58.1% for chlorogenic acid and the three minor components, indicating that these components had common structural features. Component 2 was identified as dicaffeoylquinic acid, and compounds 3 and 4 were identified as ferulic acid derivatives. The radical-scavenging activity of the Teaw extract was compared with -tocopherol, BHT and chlorogenic acid, using the DPPH and 2,2′-azinobis (3-ethylbenzothialozinesulfonic acid) radical cation (ABTS⁺) assays. The Teaw extract scavenged both free radicals more strongly than did -tocopherol and BHT, and the activity of the extract was consistent with the concentration of chlorogenic acid that was present, confirming that this component is a major contributor to the antioxidant activity. The acute toxicity of the Teaw leaf extract was investigated in mice, and it was found that the LD₅₀ of the extract was >32 g/kg. Consequently, this plant is a promising source of a natural food antioxidant.     

Chemical composition of white (Morus alba), red (Morus rubra) and black (Morus nigra) mulberry fruits

In this study, the chemical composition of white (Morus alba L.), red (Morus rubra L.) and black (Morus nigra L.) mulberry fruits grown in the East Anatolia Region of Turkey was investigated. The highest total phenolic and flavonoid contents were observed in black mulberry (1422 mg gallic acid equivalents/100 g fresh matter and 276 mg quercetin equivalents/100 g fresh matter). M. alba had the highest total fat content (1.10%), followed by M. nigra (0.95%) and M. rubra (0.85%), respectively. The major fatty acids in mulberry fruits were linoleic acid (54.2%), palmitic acid (19.8%) and oleic acid (8.41%), respectively. The total soluble solids content of mulberry species varied between 15.9% (M. rubra L.) and 20.4% (M. alba L.), acidity between 0.25% (M. alba L.) and 1.40% (M. nigra L.), pH between 3.52 (M. nigra L.) and 5.60 (M. alba L.), ascorbic acid 19.4 mg/100 g (M. rubra L.) and 22.4 mg/100 g (M. alba L.), respectively. Mineral compositions of the mulberry species were 0.83% N, 235 mg/100 g P, 1141 mg/100 g K, 139 mg/100 g Ca, 109 mg/100 g Mg, 60 mg/100 g Na, 4.3 mg/100 g Fe, 0.4 mg/100 g Cu, 4.0 mg/100 g Mn and 3.1 mg/100 g Zn, respectively.     

Phenolic Profiles,Antioxidant Activities,and Neuroprotective Properties of Mulberry (Morus atropurpurea Roxb.) Fruit Extracts from Different Ripening Stages

The present work investigated the phenolic profiles (including nonanthocyanin and anthocyanin phenolics), antioxidant activities, and neuroprotective potential of mulberry fruit (MF) (Morus atropurpurea Roxb.) grown in China at different ripening stages. High‐performance liquid chromatography‐tandem mass spectrometry method (HPLC‐MS/MS) was used to identify and quantify the phenolic compounds. The antioxidant capacity, total phenolic content (TPC), total flavonoid content (TFC), and total monomeric anthocyanin content (TAC) were determined using spectrophotometric methods. The neuroprotective effects of MFs at different ripening stages were investigated using Aβ_25‐35‐treated PC12 cells as the cellular model of Alzheimer's disease. Of the 19 phenolic compounds characterized from the MF extracts, the contents of rutin and anthocyanins increased and that of chlorogenic acid decreased significantly with maturity. At the fully ripened stage, MF extracts showed the highest amounts of TPC (11.23 mg gallic acid equivalents/g fresh weight), TFC (15.1 mg rutin equivalents/g fresh weight), and TAC (1177 mg cyanidin 3‐O‐glucoside equivalents/100 g fresh weight). Meanwhile, antioxidant activity of MF extracts at this stage was highest according to ABTS (an IC50 value of 4.11 μg/mL) and DPPH (an IC50 value of 10.08 μg/mL) assays. Cellular assays revealed increased cell viability in cells treated with the ripe MF extracts; compared with the control groups, the ripening fruits also increased the antioxidant enzyme levels in PC12 cells. Together, these results suggest that the antioxidant activities and neuroprotective properties of ripening MFs are related to the contents and types of phenolic compounds that are present in the fruits.     

Anti-oxidant capacities of flavonoid compounds isolated from acai pulp (Euterpe oleracea Mart.)

Acai fruit (Euterpe oleracea Mart.) has been demonstrated to exhibit extremely high anti-oxidant capacity. Seven major flavonoids were isolated from freeze-dried acai pulp by various chromatographic methods. Their structures were elucidated as orientin (1), homoorientin (2), vitexin (3), luteolin (4), chrysoeriol (5), quercetin (6), and dihydrokaempferol (7) by NMR, MS and compared with the reported literature. Compounds 3 and 6 were reported from acai pulp for the first time. Anti-oxidant capacities of these flavonoids were evaluated by oxygen radical absorbance capacity (ORAC) assay, cell-based anti-oxidant protection (CAP-e) assay and reactive oxygen species (ROS) formation in polymorphonuclear (PMN) cells (ROS PMN assay). ORAC values varied distinctly (1420–14,800 μmol TE/g) among the seven compounds based on numbers and positions of hydroxyl groups and/or other substitute groups. The ORAC values of aglycones are generally higher than that of glycosides. CAP-e results indicated that only three compounds (4, 6 and 7) could enter the cytosol and contribute to the reduction of oxidative damage within the cell. The ROS PMN assay showed that five compounds (2–3 and 5–7) demonstrated exceptional effects by reducing ROS formation in PMN cells, which produced high amounts of ROS under oxidative stress. In evaluating the anti-oxidant capacity of natural products, combining both chemical and cell-based assays will provide more comprehensive understanding of anti-oxidant effects and potential biological relevance.     

Star-anise (Illicium verum) and black caraway (Carum nigrum) as natural antioxidants

The various solvent fractions of star-anise (Illicium verum) and black caraway (Carum nigrum), along with their spice powders and volatile oils, were prepared and evaluated for antioxygenic activity, using different methods. Star-anise powder and its ethanol/water (80:20)-soluble fraction showed strong antioxygenic activity in refined sunflower oil while the petroleum ether fraction exhibited marginal antioxygenic activity and the water-soluble fraction was practically devoid of any activity in sunflower oil. The black caraway powder showed marginal antioxygenic activity while its ethanol/water fraction (80:20) showed strong antioxygenic activity and all other fractions showed slight pro-oxygenic activity in refined sunflower oil. Both the spice powders and their extracts were also evaluated for antioxidant activity by linoleic acid peroxidaton, β-carotene-linoleate and 1, 1-diphenyl-2-picryl hydrazyl (DPPH) methods. Both the star-anise and black caraway powders, as well as their ethanol/water extracts, exhibited strong antioxygenic activity. Volatile oils from both the spices exhibited antioxygenic activity and the activity did not seem to be concentration-dependent. Volatile oils from star-anise showed relatively higher antioxygenic activity than did those from black caraway. Gas chromatography–mass spectroscopy (GC–MS) studies on star-anise and black caraway volatile oils resulted in the identification of 25 and 22 compounds, respectively, representing 94–97% of the total content.