Influence of feeding malt, bread crust, and a pronylated protein on the activity of chemopreventive enzymes and antioxidative defense parameters in vivo

The aim of the present study was to investigate whether feeding of malt, bread crust, and a pronylated albumin modulates chemoprevention enzymes, such as glutathione-S-transferase (GST) and UDP-glucuronyl-transferase (UDP-GT), and antioxidative defense parameters in vivo and whether the intake of these foods rich in Maillard reaction compounds results in an accumulation of compounds formed in in vivo glycation reactions. After quantitation of pronylated lysine in malt and bread crust, male Wistar rats were fed a standard chow supplemented with 28% of protein containing different amounts of casein, bread crust, caraffa malt, or pronyl bovine serum albumin (BSA) for 15 days. GST activity in the kidneys was increased by 18% (p > 0.05) in animals of the bread crust group, while UDP-GT activity was elevated by 27% in the liver of animals administered pronyl-BSA. Contents of tocopherol in plasma were increased by 33, 14, and 14% in the bread crust, malt, and pronyl-BSA group compared to the control group, while the levels of thiobarbituric acid reactive substances were decreased and the total antioxidant capacity was increased. Parameters of endogenous glycation indicated a 32 and 46% higher load of advanced glycation end products in the kidneys after administration of the malt and the pronyl-BSA containing diet. However, the main systemic effects of dietary malt, bread crust, and pronyl-BSA were, for the first time, demonstrated to be the enhanced antioxidant capacity and the particulate increase in chemopreventive enzymes.     

In vitro antioxidant activity of coffee compounds and their metabolites

In this paper we report the antioxidant activity of different compounds which are present in coffee or are produced as a result of the metabolism of this beverage. In vitro methods such as the ABTS*+ [ABTS = 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)] decolorization assay and the oxygen radical absorbance capacity assay (ORAC) were used to assess the capacity of coffee compounds to scavenge free radicals. The importance of caffeine metabolites and colonic metabolites in the overall antioxidant activity associated with coffee consumption is shown. Colonic metabolites such as m-coumaric acid and dihydroferulic acid showed high antioxidant activity. The ability of these compounds to protect human low-density lipoprotein (LDL) oxidation by copper and 2,2'-azobis(2-amidinopropane) dihydrochloride was also explored. 1-Methyluric acid was particularly effective at inhibiting LDL oxidative modification. Different experiments showed that this caffeine metabolite is not incorporated into LDL particles. However, at physiologically relevant concentrations, it was able to delay for more than 13 h LDL oxidation by copper.     

Antioxidant properties of roasted coffee residues

The antioxidant activity of roasted coffee residues was evaluated. Extraction with four solvents (water, methanol, ethanol, and n-hexane) showed that water extracts of roasted coffee residues (WERCR) produced higher yields and gave better protection for lipid peroxidation. WERCR showed a remarkable protective effect on oxidative damage of protein. In addition, WERCR showed scavenging of free radicals as well as the reducing ability and to bind ferrous ions, indicating that WERCR acts as both primary and secondary antioxidants. The HPLC analyses showed that phenolic acids (chlorogenic acid and caffeic acid) and nonphenolic compounds [caffeine, trigonelline, nicotinic acid, and 5-(hydroxymethyl)furfuraldehyde] remained in roasted coffee residues. These compounds showed a protective effect on a liposome model system. The concentrations of flavonoids and polyphenolic compounds in roasted coffee residues were 8,400 and 20,400 ppm, respectively. In addition, the Maillard reaction products (MRPs) remaining in roasted coffee residues were believed to show antioxidant activity. These data indicate that roasted coffee residues have excellent potential for use as a natural antioxidant source because the antioxidant compounds remained in roasted coffee residues.     

Isolation, identification, and quantification of roasted coffee antibacterial compounds

Coffee brew is a widely consumed beverage with multiple biological activities due both to naturally occurring components and to the hundreds of chemicals that are formed during the roasting process. Roasted coffee extract possesses antibacterial activity against a wide range of microorganisms, including Staphylococcus aureus and Streptococcus mutans, whereas green coffee extract exhibits no such activity. The naturally occurring coffee compounds, such as chlorogenic acids and caffeine, cannot therefore be responsible for the significant antibacterial activity exerted by coffee beverages against both bacteria. The very low minimum inhibitory concentration (MIC) found for standard glyoxal, methylglyoxal, and diacetyl compounds formed during the roasting process points to these alpha-dicarbonyl compounds as the main agents responsible for the antibacterial activity of brewed coffee against Sa. aureus and St. mutans. However, their low concentrations determined in the beverage account for only 50% of its antibacterial activity. The addition of caffeine, which has weak intrinsic antibacterial activity, to a mixture of alpha-dicarbonyl compounds at the concentrations found in coffee demonstrated that caffeine synergistically enhances the antibacterial activity of alpha-dicarbonyl compounds and that glyoxal, methylglyoxal, and diacetyl in the presence of caffeine account for the whole antibacterial activity of roasted coffee.     

Screening of raw coffee for thiol binding site precursors using "in bean" model roasting experiments

The purpose of the following study was to investigate the influence of coffee roasting on the thiol-binding activity of coffee beverages, and to investigate the potential of various green bean compounds as precursors of thiol-binding sites by using promising "in bean" model roast experiments. Headspace gas chromatographic analysis on coffee brews incubated in the presence of the roasty-sulfury smelling 2-furfurylthiol for 20 min at 30 degrees C in septum-closed vessels revealed that the amounts of "free" thiol decreased drastically with increasing the roasting degree of the beans used for preparation of the brews. A half-maximal binding capacity (BC(50)) of 183 mg of 2-furfurylthiol per liter of standard coffee beverage was determined for a roasted coffee (CTN value of 67), thus demonstrating that enormous amounts of the odor-active thiol are "bound" by the coffee. Furthermore, biomimetic "in bean" precursor experiments have been performed in order to elucidate the precursor for the thiol-binding sites in the raw coffee bean. These experiments opened the possibility of studying coffee model reactions under quasi-natural roasting conditions and undoubtedly identified chlorogenic acids as well as thermal degradation products caffeic acid and quinic acid as important precursors for low-molecular-weight thiol-binding sites. In particular, when roasted in the presence of transition metal ions, chlorogenic acids and even more caffeic acid showed thiol-binding activity which was comparable to the activity measured for the authentic coffee brew.     

In vitro antioxidant and ex vivo protective activities of green and roasted coffee

The antioxidant properties of green and roasted coffee, in relation to species (Coffea arabica and Coffea robusta) and degree of roasting (light, medium, dark), were investigated. These properties were evaluated by determining the reducing substances (RS) of coffee and its antioxidant activity (AA) in vitro (model system beta-carotene-linoleic acid) and ex vivo as protective activity (PA) against rat liver cell microsome lipid peroxidation measured as TBA-reacting substances. RS of C. robustasamples were found to be significantly higher when compared to those of C. arabica samples (p < 0.001). AA for green coffee samples were slightly higher than for the corresponding roasted samples while PA was significantly lower in green coffee compared to that of all roasted samples (p < 0.001). Extraction with three different organic solvents (ethyl acetate, ethyl ether, and dichloromethane) showed that the most protective compounds are extracted from acidified dark roasted coffee solutions with ethyl acetate. The analysis of acidic extract by gel filtration chromatography (GFC) gave five fractions. Higher molecular mass fractions were found to possess antioxidant activity while the lower molecular mass fractions showed protective activity. The small amounts of these acidic, low molecular mass protective fractions isolated indicate that they contain very strong protective agents.     

In vitro antioxidant activities of barley, husked oat, naked oat, triticale, and buckwheat wastes and their influence on the growth and biomarkers of antioxidant status in rats

The study was aimed at verification of the following hypothesis: differences in antioxidant capacity of diets consisting of different cereals and byproducts affect the antioxidant status of the consumers of these diets. To validate that hypothesis this study investigated the contents of polyphenols and alpha-tocopherol as well as the total antioxidant capacity (TAC) in vitro of cereals and their fractions (barley, husked and naked oat, oat bran, and triticale); the nutritional and antioxidant properties of diets containing these cereals, applied in a 4-week feeding experiment on rats, were also assessed. Among the cereals examined, the highest TAC was reported for barley (13.16 micromol of Trolox/g) and the lowest for naked oat (3.84 micromol of Trolox/g). Compared with cereals, the TAC of buckwheat waste was 2-3 times higher (25.2 micromol of Trolox/g). The antioxidant capacity of diets, calculated in vitro, ranged from 6.35 micromol of Trolox/g for naked oat type diet to 10.51 micromol of Trolox/g for barley type diet. Results of an in vitro study were confirmed in changes of glutathione peroxidase (GPx) activities and the level of thiobarbituric acid-reactive substances (TBARS) in the serum of rats fed diets with the highest and lowest antioxidant capacities in vitro; the barley diet increased the activity of GPx (37.63 units/mL) and decreased the level of TBARS (4.82 microg/g), whereas the naked oat diet had an opposite effect (31.16 units/mL and 5.91 microg/g, respectively).     

Biological activity of total lipids from red and white wine/must.

Wine is an essential component of the Mediterranean diet, and it is thought to exert a protective effect against coronary heart disease. Although many efforts have been made to determine the protective compounds in wines, their exact nature and how they are involved in the protection mechanisms are still unclear. In this study, total lipids, total polar lipids, and total neutral lipids of five wines and three musts were tested in vitro for their ability to induce washed rabbit platelet aggregation and/or to inhibit platelet activating factor (PAF) induced aggregation. The results showed that the biological activity of wine/must total lipids can be attributed mainly to total polar lipids. In the red wine Cabernet Sauvignon, we fractionated total neutral lipids, total polar lipids, and pigments by HPLC. Each fraction was tested in vitro for its biological activity. Structural data of the most active fractions, based on biological, chemical, and spectral methods, are also presented.     

Increase of the activity of phase II antioxidant enzymes in rats after a single dose of coffee

This study evaluated the acute effect of the administration of coffee brew in the activity of phase II antioxidant enzymes in the hepatic tissue of rats. A single dose of this beverage increased the activity of the enzymes SOD, CAT, and GPx; the maximum increase occurred 1 h after administration (19.1, 22.1, and 25.1%, respectively). These changes were statistically significant (p < 0.05), the response was shown to be dose-dependent (p < 0.05), and the return to basal levels took >4 h from the intervention, suggesting a long-term effect. The total antioxidant capacity of the hepatic tissue also exhibited a peak 1 h after the intervention (6.5%), but the increase was not statistically significant and the response was not dose-dependent due to the low exposure to coffee. These results indicate that coffee increases the activities of antioxidant enzymes, improving protection against oxidative stress.     

Structural and functional characterization of pronyl-lysine,a novel protein modification in bread crust melanoidins showing in vitro antioxidative and phase I/II enzyme modulating activity

Application of an in vitro antioxidant assay to solvent fractions isolated from bread crust, bread crumb, and flour, respectively, revealed the highest antioxidative potential for the dark brown, ethanol solubles of the crust, whereas corresponding crumb and flour fractions showed only minor activities. To investigate whether these browning products may also act as antioxidants in biological systems, their modulating activity on detoxification enzymes was investigated as a functional parameter in intestinal Caco-2 cells. The bread crust and, in particular, the intensely brown, ethanolic crust fraction induced a significantly elevated glutathione S-transferase (GST) activity and a decreased phase I NADPH-cytochrome c reductase (CCR) activity compared to crumb-exposed cells. Antioxidant screening of Maillard-type model mixtures, followed by structure determination, revealed the pyrrolinone reductones 1 and 2 as the key antioxidants formed from the hexose-derived acetylformoin and N(alpha)-acetyl-L-lysine methyl ester or glycine methyl ester, chosen as model substances to mimic nonenzymatic browning reactions with the lysine side chain or the N terminus of proteins, respectively. Quantitation of protein-bound pyrrolinone reductonyl-lysine, abbreviated pronyl-lysine, revealed high amounts in the bread crust (62.2 mg/kg), low amounts in the crumb (8.0 mg/kg), and the absence of this compound in untreated flour. Exposing Caco-2 cells for 48 h to either synthetically pronylated albumin or purified pronyl-glycine (3) significantly increased phase II GST activity by 12 or 34%, respectively, thus demonstrating for the first time that "pronylated" proteins as part of bread crust melanoidins act as monofunctional inducers of GST, serving as a functional parameter of an antioxidant, chemopreventive activity in vitro.     

Dietary fiber content and associated antioxidant compounds in Roselle flower (Hibiscus sabdariffa L.) beverage

The beverage of Hibiscus sabdariffa flowers is widely consumed in Mexico. Polyphenols contained in plant foods are frequently associated with dietary fiber. The aim of this work is to quantify the dietary fiber, associated polyphenols, and antioxidant capacity of the Roselle flower and the beverage traditionally prepared from it and its contribution to the Mexican diet. Roselle flower contained dietary fiber as the largest component (33.9%) and was rich in phenolic compounds (6.13%). Soluble dietary fiber was 0.66 g/L in beverage, and 66% of total extractable polyphenols contained in Roselle flower passed to the beverage and showed an antioxidant capacity of 335 micromoL trolox equivalents/100 mL beverage measured by ABTS. These data suggest that Roselle flower beverage intake in the Mexican diet may contribute around 166 and 165 mg/per serving to the intake of dietary fiber and polyphenols, respectively. The health benefits from consumption of Hibiscus beverage could be of considerable benefit to the whole population.     

Effect of roasting on the antioxidant activity of coffee brews

Colombian Arabica coffee beans were roasted to give light, medium, and dark samples. Their aqueous extracts were analyzed by gel filtration chromatography, UV-visible spectrophotometry, capillary electrophoresis, and the ABTS(*)(+) assay. A progressive decrease in antioxidant activity (associated mainly with chlorogenic acids in the green beans) with degree of roasting was observed with the simultaneous generation of high (HMM) and low molecular mass (LMM) compounds possessing antioxidant activity. Maximum antioxidant activity was observed for the medium-roasted coffee; the dark coffee had a lower antioxidant activity despite the increase in color. Analysis of the gel filtration chromatography fractions showed that the LMM fraction made a greater contribution to total antioxidant activity than the HMM components.     

Comparison of the antioxidant activity of commonly consumed polyphenolic beverages (coffee, cocoa, and tea) prepared per cup serving.

In this study, the in vitro low-density lipoprotein oxidation model was used to assess the relative antioxidant activity of the polyphenolic beverages tea, coffee, and cocoa on a cup-serving basis. The beverages were prepared as 0.7-2.5% soluble coffee and 1.5-3.5% cocoa; teas (green, black, or herbal) were prepared as one tea bag infused over 5 min in 220 mL of hot water. Under these standard cup serving conditions, the antioxidant activity as determined by the lag time was in the range of 292-948 min for coffee, 217-444 min for cocoa, 186-338 min for green tea, 67-277 min for black tea, and 6-78 min for herbal tea. Addition of milk did not alter the antioxidant activity. The influence of coffee bean source and degree of roasting was further investigated. Green coffee beans of Robusta coffee exhibited a 2-fold higher antioxidant activity than Arabica coffee, but after roasting this difference was no longer significant. In conclusion, these commonly consumed beverages have a significant antioxidant activity, the highest being soluble coffee on a cup-serving basis.     

Effect of in vitro enzymatic digestion on antioxidant activity of coffee melanoidins and fractions

Traditionally antioxidant activity of melanoidins has only been evaluated in food for implication in shelf life but gastrointestinal digestion is necessary to study their potential bioactivity. In addition, the biological fate of melanoidins has been stressed during the past decade since they did not behave as inert substances. In the present paper a soluble coffee melanoidin isolated from brewed coffee after ultrafiltration with a 10 kDa cutoff membrane was treated ionically and enzymatically collecting the respective high and low molecular weight fractions. Antioxidant activity of these fractions was evaluated with five well-described assays (DPPH, ABTS, ORAC, HOSC, and FRAP) that were previously setup in a plate reader based automatized analysis. Low molecular weight compounds released from melanoidin after gastrointestinal digestion exerted the highest antioxidant activity, even higher than compounds bound ionically to melanoidins. Gastrointestinal digestion is able to modify coffee melanoidins to some extent, as hypothesized from their absolute antioxidant activities. Two options are plausible: by modifying/releasing the ionically bound compounds and/or by genesis of new more active structures from the melanoidin skeleton after enzymatic treatment.     

The antioxidant and chlorogenic acid profiles of whole coffee fruits are influenced by the extraction procedures

Commercial whole coffee fruit extracts and powder samples were analyzed for chlorogenic acids (CGA), caffeine and antioxidant activities. CGA and caffeine were characterized by LC-MS(n) and HPLC accordingly, and quantified by UV absorbance. ORAC, HORAC, NORAC, SORAC and SOAC (antioxidant capacities) were assessed. Three caffeoylquinic acids, three feruloylquinic acids, three dicaffeoylquinic acids, one p-coumaroylquinic acid, two caffeoylferuloylquinic acids and three putative chlorogenic lactones were quantified, along with a methyl ester of 5-caffeoylquinic acid (detected in one sample, the first such report in any coffee material). Multistep whole coffee fruit extracts displayed higher CGA content than single-step extracts, freeze-dried, or air-dried whole raw fruits. Caffeine in multistep extracts was lower than in the single-step extracts and powders. Antioxidant activity in whole coffee fruit extracts was up to 25-fold higher than in powders dependent upon the radical. Total antioxidant activity of samples displayed strong correlation to CGA content.     

Correlation of selected constituents with the total antioxidant capacity of coffee beverages: influence of the brewing procedure

Relationships between volatile and nonvolatile compounds and the antioxidant capacity of coffee brews prepared from commercial conventional and torrefacto roasted coffees, employing commonly used doses and prepared by four brewing procedures (filter, plunger, mocha, and espresso machine) were assessed. Significant correlations between volatile Maillard reaction products and antioxidant capacity (measured by both 2,2-diphenyl-1-picrylhydrazyl radical and redox potential methods) were not observed. Highly positive correlations between browned compounds and caffeine with both antioxidant capacity parameters were reported. Principal component analysis allowed coffee brews separation according to coffee roasting processes (PC1) and brewing procedures (PC2), showing that in all cases coffee brews from torrefacto roasted coffee were more antioxidant that those extracted from conventional ones; also, coffee brews extracted by an espresso machine were more antioxidant than those extracted by mocha, plunger, and filter machines.     

Hypocholesterolemic effects of a flavonoid-rich extract of Hypericum perforatum L. in rats fed a cholesterol-rich diet

In a previous study, a flavonoid-rich extract of Hypericum perforatum L. (FEHP) was prepared and its antioxidant activity was determined by a series of models in vitro. In this study, the hypocholesterolemic effects of FEHP in rats fed a cholesterol-rich diet were tested. Forty Wistar rats fed a standard laboratory diet or a cholesterol-rich diet for 16 weeks were used. The serum lipid levels, as well as malondialdehyde (MDA) and activity of superoxide dismutase (SOD) and catalase (CAT) in serum and liver, were examined. Cholesterol-rich diet induced hypercholesterolemia was manifested in the elevation of serum lipid levels such as total cholesterol (TC), total triglycerides (TG), and low density lipoprotein cholesterol (LDL-C). Administration of middle-dose (75 mg/kg of BW/day) and high-dose (150 mg/kg of BW/day) FEHP significantly lowered the serum levels of TC, TG, and LDL-C, while increasing the serum level of high density lipoprotein cholesterol (HDL-C). Also, the content of MDA in serum and liver decreased significantly after oral administration of FEHP compared with those of rats fed a cholesterol-rich diet. In addition, FEHP increased the activity of SOD in serum and liver, but the activity of CAT was significantly elevated only in liver. These results suggested that the hypocholesterolemic effects of FEHP might be due to its abilities to lower serum TC, TG, and LDL-C levels as well as to slow the lipid peroxidation process and to enhance the antioxidant enzyme activity.     

Lipid fractions with aggregatory and antiaggregatory activity toward platelets in fresh and fried cod (Gadus morhua): correlation with platelet-activating factor and atherogenesis

Cod (Gadus morhua) is a popular part of the diet in many countries on both sides of the North Atlantic; in most cases it is consumed fried. In this study, total lipids of cod muscle were separated into neutral and polar lipids, which were further fractionated by HPLC. The lipid fractions were tested in vitro, against washed rabbit platelets, for the probable existence of lipid compounds that either exhibit an action similar to that of platelet-activating factor (PAF) or inhibit the action of PAF. The platelet bioassay was used to evaluate total lipids, total polar lipids, and total neutral lipids, before any further separation. Detection of these compounds in fresh and fried cod could be used to evaluate the nutritional value of this important fish. The in vitro biological study of lipids showed that in fresh cod lipid fractions, ranges of PAF-like and anti-PAF-like activities were present, whereas in fried cod lipid fractions, both neutral and polar, anti-PAF activities were mainly observed. Because it has already been reported that PAF is involved in atheromatosis generation, the existence of PAF inhibitors in cod may contribute to the possible protective role of fish, in this case cod, against atherosclerosis.     

Antioxidant and chemopreventive properties of polyphenolic compounds derived from Greek legume plant extracts

Recently, phytochemical compounds present in legumes have gained a lot of interest because they are considered to be possible chemopreventive agents. In the present study, 14 polyphenolic compounds were extracted and identified from two unique varieties of Leguminosae family plants cultivated in Greece and screened for their antioxidant and chemopreventive properties. Ten polyphenolic fractions, which are mainly mixtures of two compounds and five pure flavonoids, were isolated from the methanolic extracts of aerial plant parts of Vicia faba and Lotus edulis (Leguminosae), respectively. All of these fractions exhibited significant DPPH(*) radical scavenging capacity. Furthermore, they exerted significant protective activity against free radical-induced DNA damage. This activity was more potent against ROO(*) radical-induced DNA damage than against that induced by OH(*) radicals. Finally, they exhibited significant ability to inhibit the activity of the topoisomerase I enzyme. These results imply that the polyphenolic compounds identified in the fractions were responsible of the observed properties of the fractions and the initial extracts and indicate different mechanisms by which these phenolic compounds may act as chemopreventive agents.     

Studies on the antioxidant activities of natural vanilla extract and its constituent compounds through in vitro models

Vanilla extract was prepared by extraction of cured vanilla beans with aqueous ethyl alcohol (60%). The extract was profiled by HPLC, wherein major compounds, viz., vanillic acid, 4-hydroxybenzyl alcohol, 4-hydroxy-3-methoxybenzyl alcohol, 4-hydroxybenzaldehyde and vanillin, could be identified and separated. Extract and pure standard compounds were screened for antioxidant activity using beta-carotene-linoleate and DPPH in vitro model systems. At a concentration of 200 ppm, the extract showed 26% and 43% of antioxidant activity by beta-carotene-linoleate and DPPH methods, respectively, in comparison to corresponding values of 93% and 92% for BHA. Interestingly, 4-hydroxy-3-methoxybenzyl alcohol and 4-hydroxybenzyl alcohol exhibited antioxidant activity of 65% and 45% by beta-carotene-linoleate method and 90% and 50% by DPPH methods, respectively. In contrast, pure vanillin exhibited much lower antioxidant activity. The present study points toward the potential use of vanilla extract components as antioxidants for food preservation and in health supplements as nutraceuticals.