Antifungal activity of a de novo synthetic peptide and derivatives against fungal food contaminants.

The development of novel solutions to fight microbial food contaminants rests upon two pillars, which are the development of resistant strains and consumers' desire for a reduced consumption of synthetic drugs. Natural antimicrobial peptides possess the qualities to overcome these issues. De novo synthesis of novel antifungal compounds is a major progress that has been facilitated by the identification of parameters involved in the antimicrobial activity. A 14-residue peptide named KK14, with the sequence KKFFRAWWAPRFLK-NH2 , was designed and inhibited conidial germination and fungal growth of food contaminants within the range 6.25 to 50 µg/ml and 6.25 to 100 µg/ml, respectively. The study of three analogues of the peptide highlighted the role of some residues in the structural conformation of the peptide and its antifungal activity. The substitution of a Pro residue with Arg increased the helical content of the peptide not only its antifungal activity but also its cytotoxicity. The insertion of an unnatural bulky residue ß-diphenylalanine or a full d-enantiomerization overall increased the antifungal potency. The four peptides showed similar behaviour towards salt increase, heat treatment, and pH decrease. Interestingly, the denantiomer remained the most active at high pH and after proteolytic digestion. The four peptides did not present haemolytic activity up to 200 µg/ml but had different behaviours of cytotoxicity. These differences could be crucial for potential application as pharmaceutical or food preservatives.

Cereal type significantly affects the composition and reconstitution characteristics of dried fermented milk-cereal composites.

BACKGROUND: Dairy and cereal products are frequently combined to create composites with enhanced nutritional benefits. Commercially available dried dairy-cereal composites are typically reconstituted and cooked to produce porridge or soup. RESULTS: Dried fermented milk-cereal composites (FMCC) with ∼193 g kg-1 protein were prepared by blending fermented milk with parboiled oats (FMCCo), wheat (FMCCw), or barley (FMCCb), incubating the blend, drying, and milling. Cereal type significantly affected the composition of the FMCC and the properties of the reconstituted, cooked FMCC (R-FMCC). The FMCCo had a higher starch and fat content and lower levels of lactose, lactic acid, and amylose than FMCCb. The R-FMCCo had higher viscosity during cooking at 95 °C and cooling to 35 °C, and higher values of yield stress (σ0 ), consistency index (K) and viscosity on shearing from 20 to 120 s-1 at 60 °C than R-FMCCb. The FMCCw had lower levels of fat and ß-glucan than FMCCo or FMCCb, but was otherwise closer to FMMCb with respect to composition, cooking properties and flow behavior. CONCLUSION: Differences in composition and consistency associated with cereal type are likely to affect the nutritional value of the FMCC. © 2018 Society of Chemical Industry.

Antioxidant, immunomodulatory and antiproliferative effects of gelatin hydrolysate from unicorn leatherjacket skin.

BACKGROUND: The in vitro cellular bioactivities including, antioxidant, immunomodulatory and antiproliferative effects of a gelatin hydrolysate (GH) prepared from unicorn leatherjacket skin, using partially purified glycyl endopeptidase, were investigated in order to optimize the use of fish skin waste products as functional food ingredients. RESULTS: GH under the tested concentrations (750-1500 µg mL(-1) ) protected against H2 O2 -induced DNA damage in U937 cells. GH also protected against the H2 O2 -induced reduction in cellular antioxidant enzyme activities, superoxide dismutase and catalase, in HepG2 cells. GH demonstrated immunomodulatory potential by reducing pro-inflammatory cytokine (interleukin-6 (IL-6) and IL-1ß) production and nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. Cell proliferation in human colon cancer (Caco-2) cells was significantly reduced in a dose-dependent manner following incubation with GH. CONCLUSION: These results indicate that GH has several bioactivities which support its potential as a promising functional food ingredient with various health benefits. © 2015 Society of Chemical Industry.

Antioxidant activities and selected characteristics of gelatin hydrolysates from seabass (Lates calcarifer) skin as affected by production processes.

Antioxidant activities and selected characteristics of gelatin hydrolysates from seabass skin as affected by production processes were investigated. Hydrolysates were prepared using different processes, including hydrolysis during and after gelatin extraction. Samples hydrolysed during gelatin extraction showed a higher degree of hydrolysis (DH) and yield compared with those hydrolysed after gelatin extraction (p < 0.05). All hydrolysates had a creamy yellowish colour. A lower abundance of volatile compounds was found in the hydrolysates produced during gelatin extraction, in comparison with those obtained after gelatin extraction. Hydrolysates prepared during gelatin extraction had higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferric reducing antioxidative power (FRAP) and ferrous ion chelating activity (p < 0.05). Following a simulated in vitro gastrointestinal digestion, the DPPH radical scavenging activity and FRAP of the hydrolysates was retained, whilst ferrous ion chelating activity increased. The most appropriate conditions for the generation of antioxidant hydrolysates from seabass skin were identified.

A study of the ability of bioactive extracts from brewers' spent grain to enhance the antioxidant and immunomodulatory potential of food formulations following in vitro digestion.

Bioactivity of a snack-bar, chocolate-drink and yogurt fortified with brewers' spent grain (BSG) phenolic extracts (P2 or B2) or protein hydrolysates (barley protein hydrolysate (BPH), BPH < 3 kDa, BPH < 5 kDa, BPH > 5 kDa) was measured following gastrointestinal in vitro digestion. Concentrations of 0.5 and 0.1% (v/v) digestates were chosen for addition to Caco-2 and Jurkat T cells, respectively. Yogurt and B2 digestate protected against H2O2-induced DNA damage in Caco-2 cells (p < 0.05), by the comet assay. Snack-bar digestates possessed significant (p < 0.05) immunomodulatory effects, measured by ELISA in concanavalin-A stimulated Jurkat T cells. Addition of BPH enhanced (p < 0.05) the IFN-γ reducing capacity of the snack-bar while addition of BPH < 3 and < 5 kDa reduced IL-2 production to a greater extent than unfortified yogurt (p < 0.05). Selected BSG components can enhance the antioxidant and anti-inflammatory potential of foods.

Phenolic-enriched fractions from brewers' spent grain possess cellular antioxidant and immunomodulatory effects in cell culture model systems.

BACKGROUND: Large quantities of brewers' spent grain (BSG), a co-product of the brewing industry, are produced annually. BSG contains hydroxycinnamic acids, and phenolic-rich extracts from BSG have previously demonstrated the ability to protect against oxidant-induced DNA damage. The present study investigated the anti-inflammatory potential of eight phenolic extracts from BSG: four pale (P1-P4) and four black (B1-B4) extracts. RESULTS: BSG extracts were more cytotoxic in Jurkat T than U937 cells, with lower IC50 values in Jurkat T cells, measured using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Pale BSG extracts P2 and P3 showed the greatest anti-inflammatory potential, significantly (P < 0.05) reducing interleukin-2 (IL-2), interleukin-4 (IL-4, P2 only), interleukin-10 (IL-10) and interferon-γ (IFN-γ) production. In addition, extracts P1-P3 and B2-B4 showed significant (P < 0.05) antioxidant effects, determined by the cellular antioxidant activity assays superoxide dismutase, catalase and glutathione content (GSH). CONCLUSION: Phenolic extracts from BSG, particularly the pale BSG extracts, have the ability to reduce a stimulated cytokine production and may also protect against cellular oxidative stress. Results of the present study highlight the potential of BSG phenolic extracts to act as functional food ingredients, providing an alternative use and improving the value of this brewing industry co-product.

Development of a dehydrated fortified food base from fermented milk and parboiled wheat, and comparison of its composition and reconstitution behavior with those of commercial dried dairy-cereal blends.

Dehydrated blends of milk and cereal are reconstituted and consumed as a nutritious soup or porridge in many regions; the composition and reconstitution behavior of the blends are likely to impact on nutritional quality and consumer acceptability of the soup/porridge. Experimental samples of dried fermented milk-bulgur wheat blend (FMBW) and commercial samples of dried dairy-cereal blends, namely kishk, tarhana, and super cereal plus corn-soy blend (SCpCSB) were compared for composition, color, water sorption, and reconstitution characteristics. FMBW blends had higher contents of protein, Ca, lactose and lactic acid, lower levels of salt (NaCl) and Fe, and a lighter, more-yellow color (higher L* and b*-color co-ordinates) than tarhana or kishk. Compared with SCpCSB, FMBW had numerically higher levels of protein, lactose, and lactic acid, lower levels of Ca, Fe, Zn, and Mg, and lower pH. Tarhana had highest mean levels of starch, and on reconstitution (133 g/kg) had highest water holding capacity, viscosity during pasting and cooling, yield stress (σ 0), consistency coefficient (K), and viscosity on shearing from 20 to 120 s-1 at 60°C. Reconstituted FMBW, kishk, and SCpCSB had similar pasting and flow behavior properties. Overall, the composition (starch, protein, Ca, Mg), pasting and flow behavior characteristics of FMBW were closer to those SCpCSB and kishk than to tarhana. The results suggest that the FMBW powder, on appropriate supplementation with Ca, Fe, Zn and Mg, could be used for the development of customized fortified blended foods for specific groups.

Fortified Blended Food Base: Effect of Co-Fermentation Time on Composition, Phytic Acid Content and Reconstitution Properties.

Dehydrated blends of dairy-cereal combine the functional and nutritional properties of two major food groups. Fortified blended food base (FBFB) was prepared by blending fermented milk with parboiled wheat, co-fermenting the blend at 35 °C, shelf-drying and milling. Increasing co-fermentation time from 0 to 72 h resulted in powder with lower lactose, phytic acid and pH, and higher contents of lactic acid and galactose. Simultaneously, the pasting viscosity of the reconstituted base (16.7%, w/w, total solids) and its yield stress (σ0), consistency index (K) and viscosity on shearing decreased significantly. The changes in some characteristics (pH, phytic acid, η120) were essentially complete after 24 h co-fermentation while others (lactose, galactose and lactic acid, pasting viscosities, flowability) proceeded more gradually over 72 h. The reduction in phytic acid varied from 40 to 58% depending on the pH of the fermented milk prior to blending with the parboiled cereal. The reduction in phytic acid content of milk (fermented milk)-cereal blends with co-fermentation time is nutritionally desirable as it is conducive to an enhanced bioavailability of elements, such as Ca, Mg, Fe and Zn in milk-cereal blends, and is especially important where such blends serve as a base for fortified-blended foods supplied to food-insecure regions.

Comparison of the nutritional composition of experimental fermented milk:wheat bulgur blends and commercially available kishk and tarhana products.

Dried, fermented blends of dairy products and cereals, such as kishk and tarhana, are foodstuffs traditionally consumed in many regions as they possess good nutritional qualities and extended storage stability. This study examined the nutritional composition of kishk or tarhana type products and compared with experimental blends of fermented milk and wheat bulgur containing 60-80% milk. The blends with higher milk contents had levels of protein (18.9%) and fat (5.8%) at the concentrations specified in fortified blended foods as outlined by the World Food Program. Higher milk contents were also associated with higher contents of calcium (323.2 mg/100 g), phosphorus (335.3 mg/100 g), vitamin A (486.7 µg/100 g) and α-tocopherol (174.5 µg/100 g). The nutritional content of the experimental fermented milk:wheat bulgur blends compared favourably with that of the commercial samples. These blends may be suitable as base products, to be fortified with micronutrients, for the development of fortified blended foods (FBFs) for humanitarian distribution.

The Proportion of Fermented Milk in Dehydrated Fermented Milk⁻Parboiled Wheat Composites Significantly Affects Their Composition, Pasting Behaviour, and Flow Properties on Reconstitution.

Dairy and cereal are frequently combined to create composite foods with enhanced nutritional benefits. Dehydrated fermented milk⁻wheat composites (FMWC) were prepared by blending fermented milk (FM) and parboiled wheat (W), incubating at 35 °C for 24 h, drying at 46 °C for 48 h, and milling to 1 mm. Increasing the weight ratio of FM to W from 1.5 to 4.0 resulted in reductions in total solids (from 96 to 92%) and starch (from 52 to 39%), and increases in protein (15.2⁻18.9%), fat (3.7⁻5.9%), lactose (6.4⁻11.4%), and lactic acid (2.7⁻4.2%). FMWC need to be reconstituted prior to consumption. The water-holding capacity, pasting viscosity, and setback viscosity of the reconstituted FMWC (16.7% total solids) decreased with the ratio of FM to W. The reconstituted FMWC exhibited pseudoplastic flow behaviour on shearing from 18 to 120 s-1. Increasing the FM:W ratio coincided with a lower yield stress, consistency index, and viscosity at 120 s-1. The results demonstrate the critical impact of the FM:W ratio on the composition, pasting behavior, and consistency of the reconstituted FMWC. The difference in consistency associated with varying the FM:W ratio is likely to impact on satiety and nutrient value of the FMWCs.

Modulatory effects of resveratrol, citroflavan-3-ol, and plant-derived extracts on oxidative stress in U937 cells.

Phytochemicals and plant extracts, present in fruit, vegetables, plants, herbs, and beverages, have been shown to have antioxidant potential that may modulate the etiology of certain chronic diseases. The objective of the present study was to determine the concentration of compound that inhibited cell growth by 50% (IC(50)) of a range of phytochemicals and plant extracts and to investigate their antioxidant and genoprotective effects under conditions of oxidative stress in U937 cells. Two phytochemicals-resveratrol and citroflavan-3-ol-and four plant extracts-grapeseed polyphenols, olive leaf extract, bearberry, and Echinacea purpurea-were examined. Viability was assessed by the fluorescein diacetate/ethidium bromide assay. The IC(50) was calculated. To examine their antioxidant and genoprotective effects, U937 cells were pretreated with the test compounds at levels below the IC(50) and then exposed to oxidants: 0.5 microM etoposide or 100 microM hydrogen peroxide (H(2)O(2)) or 400 microM tert-butylhydroperoxide (tBOOH). Cellular reduced glutathione levels were measured as an indicator of oxidative stress. DNA damage was assessed by the alkaline single-cell gel electrophoresis assay or comet assay. Resveratrol demonstrated the highest IC50 value of 13.7 microg/mL, with Echinacea the lowest at 9,400 microg/mL. Etoposide-induced oxidative stress was strongly reduced by olive leaf extract and bearberry. Grapeseed polyphenols and bearberry strongly protected against H2O2- and tBOOH-induced DNA damage. In conclusion, these results provide evidence that non-nutrient dietary constituents may act as significant bioactive compounds and that plant extracts, such as bearberry, grapeseed polyphenols, and olive leaf extract, strongly protect against oxidative stress.

Cellular Transport and Bioactivity of a Major Saffron Apocarotenoid, Picrocrocin (4-(ß-D-Glucopyranosyloxy)-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde).

The cellular transport and bioactivity of the second major saffron apocarotenoid, picrocrocin, was examined in parallel to that of the major group, crocetin sugar esters, in aqueous extracts. The transport of pure picrocrocin was investigated in comparison to that of other saffron apocarotenoids, trans-crocetin (di-ß-D-gentiobiosyl) ester and crocetin using the Caco-2 cell model coupled with an in vitro digestion procedure. RP-HPLC-DAD was employed to quantify the bioaccessible and bioavailable amounts of individual apocarotenoids. Picrocrocin and crocetin sugar esters though highly bioaccessible (75% and 60%, respectively) were transported at minute quantities (0.2% and 0.5%, respectively; 10-fold lower than crocetin). Picrocrocin did not protect against oxidant-induced DNA damage in U937, human monocytic blood cells at the concentration investigated, however, it reduced the proliferation of human adenocarcinoma and hepatocarcinoma cells. Our findings may be useful for the requirements of food legislation regarding saffron preparations, in which both apocarotenoid groups coexist.

Anti-inflammatory effects of wild Irish mushroom extracts in RAW264.7 mouse macrophage cells.

Mushrooms and mushroom extracts have traditionally been used as therapies for a wide variety of ailments, including allergy, arthritis, and other inflammatory disorders. However, more evidence is required on the mechanism by which mushrooms exert these effects. In the present study, the anti-inflammatory properties of ethanol and hot water extracts prepared from 27 fungal samples collected between October and November 2011 at various forest locations in the southwest of Ireland were investigated using the lipopolysaccharide (LPS)-stimulated mouse macrophage (RAW264.7 cells) model of inflammation. LPS-stimulated cells were incubated in the presence of mushroom extracts at nontoxic concentrations for 24 h and the production of interleukin-6 (IL-6) was quantified by ELISA. Seven ethanolic and one hot water extract that decreased IL-6 production were selected for further study. The extracts were then incubated with LPS-stimulated cells for 24 h and the production of IL-6, tumor necrosis factor-alpha (TNF-α), and nitric oxide (NO) was measured. Ethanolic extracts prepared from Russula mairei, Lactarius blennius, Craterellus tubaeformis, Russula fellea, and Craterellus cornucopioides demonstrated selective anti-inflammatory activity by decreasing the production of NO and IL-6 but not TNF-α in LPS-stimulated RAW264.7 cells. These findings support existing evidence of the anti-inflammatory potential of mushroom extracts.

Antioxidant and pro-apoptotic effects of marine-derived, multi-mineral aquamin supplemented with a pine bark extract, Enzogenol, and a green tea extract, Sunphenon.

A high dietary intake of polyphenols has been associated with a decreased risk of cardiovascular disease and cancer, attributed in part to their antioxidant activity and pro-apoptotic effects. Aquamin is a multi-mineral algal extract that enhances bone mineralization, relieves osteoarthritis, and aids digestion; however, Aquamin has not demonstrated antioxidant activity. In the present study, Aquamin was supplemented with 8% Enzogenol, a pine bark extract with a high phenolic content, and 2% Sunphenon, a green tea extract that also has a high phenolic content to produce a mixed product (A:E:S). The antioxidant activity of A:E:S was compared with that of its constituent compounds and also with catechin and epigallocatechin by measuring total phenol content, ferric-reducing antioxidant potential, and 2,2-diphenyl-2-picrylhydrazyl hydrate. The cytotoxic and apoptotic effects of the compounds were also measured in the U937, human monocytic blood cell line. A:E:S demonstrated an antioxidant activity that was equal to that of the compounds used in its preparation. Aquamin was not cytotoxic in the U937 cell line; however, A:E:S was cytotoxic and the primary mechanism of cell death was apoptosis. The biological effects of Aquamin were enhanced by supplementation with Enzogenol and Sunphenon to include antioxidant effects and the ability to induce apoptosis in U937 cells.

Changes in total and individual crocetin esters upon in vitro gastrointestinal digestion of saffron aqueous extracts.

Changes that may be expected in crocetin esters (crocins) upon digestion were examined in saffron aqueous extracts for the first time. Chemical characterization of total and individual crocins and other bioactive compounds was achieved by UV-vis spectrophotometry, RP-HPLC-DAD, and LC-ESI-MS. Antioxidant activity was evaluated using in vitro assays and the comet assay. The observed loss for both total and trans-crocins was higher in saffron (∼50%) than in gardenia extracts (∼30%), which were also examined for comparison. Loss was lower than that reported for hydrophobic carotenoids. cis-Isomers were less affected, leading to the hypothesis that trans/cis isomerization may occur in parallel to degradation reactions. Monitoring changes in the extracts at oral, gastric, or intestinal phases, separately, verified this view pointing out the critical effect of pH, temperature, and duration of process but not of digestive enzymes. No isomerization and less degradation (<20% loss) was evidenced when pure trans-crocetin (di-ß-D-gentiobiosyl) ester was subjected to gastric or intestinal conditions.

Brewers' spent grain; bioactivity of phenolic component, its role in animal nutrition and potential for incorporation in functional foods: a review.

Brewers' spent grain (BSG) is a low-value co-product of the brewing industry produced in large quantities annually. This paper reviews the existing evidence regarding the phenolic component of BSG, focusing on composition, extraction and biofunctions such as antioxidant, anti-atherogenic, anti-inflammatory and anti-carcinogenic activities. Furthermore, the incorporation of BSG in foodstuffs will be discussed, including the use of BSG as an animal feed supplement and the potential of BSG to be incorporated into foods for human consumption. BSG contains hydroxycinnamic acids including ferulic acid, p-coumaric acid and caffeic acid; which have shown bioactivity in the pure form (antioxidant, anti-inflammatory, anti-atherogenic and anti-cancer). Phenolic extracts from BSG have also shown antioxidant potential, by protecting against oxidant-induced DNA damage, possibly by Fe chelation. Studies show that BSG has many benefits when used as a supplement in animal feed, such as increasing milk yield, milkfat content and in providing essential dietary amino acids. The incorporation of BSG in human foods such as cookies and ready-to-eat snacks has resulted in increased protein and fibre contents of the products, where the changes in organoleptic properties are controllable. It can be concluded that the phenolic component of BSG has potential bioactive effects, which are worth pursuing given that the inclusion of BSG into human foodstuffs is viable and beneficial.

Brewers' spent grain (BSG) protein hydrolysates decrease hydrogen peroxide (H2O2)-induced oxidative stress and concanavalin-A (con-A) stimulated IFN-γ production in cell culture.

The present study investigated the bioactivity of protein hydrolysates and fractionated hydrolysates prepared from brewers' spent grain (BSG) using proteases, including Alcalase 2.4L, Flavourzyme and Corolase PP. Hydrolysates were designated K-Y, including fractionated hydrolysates with molecular weight (m.w.) < 3, <5 and >5 kDa. Where computable, IC50 values were lower in U937 (1.38-9.78%) than Jurkat T cells (1.15-13.82%). Hydrolysates L, Q and R and fractionated hydrolysates of U and W (<3, <5, >5 kDa) significantly (P < 0.01) protected against hydrogen peroxide (H2O2)-induced reduction of superoxide dismutase (SOD) activity. A fractionated hydrolysate of W (<5 kDa) protected against H2O2-induced DNA damage, P < 0.01. Hydrolysates K, N, P, U, U > 5 kDa, V, V > 5 kDa, W, W > 5 kDa significantly (P < 0.05) reduced a concanavlin-A (con-A) stimulated production of interferon-γ (IFN-γ). In conclusion, BSG protein hydrolysates demonstrate bioactivity in vitro; lower m.w. hydrolysates (<3, <5 kDa) show greatest antioxidant activity and unfractionated or higher m.w. hydrolysates (>5 kDa) possess anti-inflammatory effects.

The hydroxycinnamic acid content of barley and brewers' spent grain (BSG) and the potential to incorporate phenolic extracts of BSG as antioxidants into fruit beverages.

The hydroxycinnamic acid (HA) content of starting barley for brewers' spent grains (BSG), whole BSG and phenolic extracts from BSG was measured using high performance liquid chromatography (HPLC) and correlated with antioxidant potential. The effect of BSG phenolic extracts on antioxidant activity of fruit beverages was also assessed (using the total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays). The concentration of HA present in barley extract and BSG was in the order of ferulic acid (FA), p-coumaric acid (p-CA) derivatives, FA derivatives, p-CA, caffeic acid (CA) and CA derivatives. Results suggested that brewing and roasting decreased the HA content. Antioxidant activity was significantly (P<0.05) correlated with caffeic acid (R(2)=0.8309) and total HA (R(2)=0.3942) concentrations. Addition of extracts to fruit beverages resulted in a significant (P<0.05) increase in antioxidant activity of cranberry juice, measured by the FRAP assay. In vitro digestion significantly (P<0.05) reduced TPC, DPPH and FRAP activity of the fruit beverages.

Phenolic extracts of brewers' spent grain (BSG) as functional ingredients - assessment of their DNA protective effect against oxidant-induced DNA single strand breaks in U937 cells.

Brewers' spent grain (BSG), a by-product of the brewing industry, contains high amounts of phenolic acids, which have antioxidant effects. The present study examined the ability of BSG extracts to protect against the genotoxic effects of oxidants, hydrogen peroxide (H(2)O(2)), 3-morpholinosydnonimine hydrochloride (SIN-1), 4-nitroquinoline 1-oxide (4-NQO) and tert-butylhydroperoxide (t-BOOH) in U937 cells. Four pale (P1-P4) and four black (B1-B4) BSG extracts were investigated. U937 cells were pre-incubated with BSG extracts, exposed to the oxidants and the DNA damage was measured by the Comet assay. The black BSG extracts (B1-B4) significantly protected against H(2)O(2)-induced DNA damage. Extract B2, which had the highest phenol content, provided the greatest protection. Extracts P2, B2, B3 and B4 provided significant protection against SIN-1-induced DNA damage. None of the extracts protected against DNA damage induced by t-BOOH and 4-NQO. The DNA protective effects of the BSG phenolic extracts may be related to iron chelation.

Cytotoxic and apoptotic effects of the oxidized derivatives of stigmasterol in the U937 human monocytic cell line.

Dietary exposure to phytosterols has increased in recent years due to the incorporation of these compounds into cholesterol-lowering products. Previous studies have investigated the cytotoxic effects of the oxidized derivatives of ß-sitosterol and determined that phytosterol oxidation products (POP) have a similar but less potent toxicity compared to their cholesterol equivalents. In the present study, the cytotoxicity of the oxidized derivatives of stigmasterol were investigated in the U937 cell line. The stigmasta-5,22-diene-3ß,7ß-diol (7ß-OH), 5,6-epoxystigmasta-22,23-diol (epoxydiol), 5,6,22,23-diepoxystigmastane (diepoxide), and (22R,23R)-stigmast-5-ene-3ß,22,23-triol (22R,23R-triol) derivatives were identified as the most cytotoxic, and the mode of cell death was identified as apoptosis in cells incubated with 7ß-OH, epoxydiol, and diepoxide stigmasterol. The antioxidants α-tocopherol, γ-tocopherol, and ß-carotene did not protect against apoptosis induced by 7ß-OH and diepoxide stigmasterol; however, α-tocopherol was found to protect against epoxydiol-induced apoptosis. The cellular antioxidant, glutathione, was depleted and the apoptotic protein, Bcl-2, was down-regulated by the stigmasterol oxides identified as apoptotic.