Echinacea species and alkamides inhibit prostaglandin E(2) production in RAW264.7 mouse macrophage cells

Inhibition of prostaglandin E(2) (PGE(2)) production in lipopolysaccharide-stimulated RAW264.7 mouse macrophage cells was assessed with an enzyme immunoassay following treatments with Echinacea extracts or synthesized alkamides. Results indicated that ethanol extracts diluted in media to a concentration of 15 microg/mL from E. angustifolia, E. pallida, E. simulata, and E. sanguinea significantly inhibited PGE2 production. In further studies, PGE2 production was significantly reduced by all synthesized alkamides assayed at 50 microM, by Bauer alkamides 8, 12A analogue, and 14, Chen alkamide 2, and Chen alkamide 2 analogue at 25 microM and by Bauer alkamide 14 at 10 microM. Cytotoxicity did not play a role in the noted reduction of PGE2 production in either the Echinacea extracts or synthesized alkamides. High-performance liquid chromatography analysis identified individual alkamides present at concentrations below 2.8 microM in the extracts from the six Echinacea species (15 microg/mL crude extract). Because active extracts contained <2.8 microM of specific alkamide and the results showed that synthetic alkamides must have a minimum concentration of 10 microM to inhibit PGE2, it is likely that alkamides may contribute toward the anti-inflammatory activity of Echinacea in a synergistic or additive manner.     

Synergistic antioxidative effects of alkamides, caffeic acid derivatives, and polysaccharide fractions from Echinacea purpurea on in vitro oxidation of human low-density lipoproteins

Preparations of Echinacea are widely used as alternative remedies to prevent the common cold and infections in the upper respiratory tract. After extraction, fractionation, and isolation, the antioxidant activity of three extracts, one alkamide fraction, four polysaccharide-containing fractions, and three caffeic acid derivatives from Echinacea purpurea root was evaluated by measuring their inhibition of in vitro Cu(II)-catalyzed oxidation of human low-density lipoprotein (LDL). The antioxidant activities of the isolated caffeic acid derivatives were compared to those of echinacoside, caffeic acid, and rosmarinic acid for reference. The order of antioxidant activity of the tested substances was cichoric acid > echinacoside > or = derivative II > or = caffeic acid > or = rosmarinic acid > derivative I. Among the extracts the 80% aqueous ethanolic extract exhibited a 10 times longer lag phase prolongation (LPP) than the 50% ethanolic extract, which in turn exhibited a longer LPP than the water extract. Following ion-exchange chromatography of the water extract, the majority of its antioxidant activity was found in the latest eluted fraction (H2O-acidic 3). The antioxidant activity of the tested Echinacea extracts, fractions, and isolated compounds was dose dependent. Synergistic antioxidant effects of Echinacea constituents were found when cichoric acid (major caffeic acid derivative in E. purpurea) or echinacoside (major caffeic acid derivative in Echinacea pallida and Echinacea angustifolia) were combined with a natural mixture of alkamides and/or a water extract containing the high molecular weight compounds. This contributes to the hypothesis that the physiologically beneficial effects of Echinacea are exerted by the multitude of constituents present in the preparations.     

Studies on the antioxidant activity of Echinacea root extract

Methanol extracts of freeze-dried Echinacea (E. angustifolia, E. pallida, and E. purpurea) roots were examined for free radical scavenging capacities and antioxidant activities. Root extracts of E. angustifolia, E. pallida, and E. purpurea were capable of scavenging hydroxyl radical. Similar scavenging activities for each variety were found for both 1,1-diphenyl-2-picrylhydrazyl radical and ABTS radical. Meanwhile, antioxidant activities of all three varieties of Echinacea were found to delay the formation of conjugated diene hydroperoxide induced by the thermal decomposition of 2, 2'-azobis(2-amidinopropane) dihydrochloride and extend the lag phase of peroxidation of soybean liposomes. Echinacea root extracts suppressed the oxidation of human low-density lipoprotein, as evaluated by reduced agarose electrophoretic mobility following oxidative modification by Cu(2+). The mechanisms of antioxidant activity of extracts derived from Echinacea roots included free radical scavenging and transition metal chelating.     

Human gene expression as a tool to determine horticultural maturity in a bioactive plant (Echinacea purpurea L. Moench)

A phenological study was conducted to determine the impact of harvest maturity on the immune-modulating properties of Echinacea purpurea. The aerial parts of this plant were collected during seven stages of development and were assayed for a common botanical marker for this species, cichoric acid. Plants of selected development stages were also assayed for total polysaccharides and compared for their immune-modulating effects on the THP-1 monocyte/macrophage cell line by means of a gene expression study. Although the concentration of cichoric acid did not change significantly during the course of the study, stage 1 (advanced vegetative) had the highest concentration of total polysaccharides and exhibited the most potent induction activity on immune-modulating cytokines such as interferon-gamma and tumor necrosis factor-alpha. These findings suggest that the use of gene expression may be an effective tool not only to standardize botanical extracts but also to optimize harvest time.     

Phytochemical variation in echinacea from roots and flowerheads of wild and cultivated populations

Quantitative phytochemical variation was determined from roots and inflorescences of native plant populations in the genus Echinacea. Specimens were collected in situ throughout the natural range of each putative taxon and transplanted to greenhouse cultivation. Ethanolic extracts from individual plants were separated by reversed-phase HPLC to quantify the alkamides, polyenes/ynes, and phenolics, and then grouped by age and taxonomically, according to a recent morphometric taxonomic revision of the genus. Canonical discriminant analysis revealed that cichoric acid, the diene alkamides 1-3 and 7, and ketoalkene 24 were the best taxonomic markers. Mean content for each of 26 phytochemicals revealed useful agronomic information, such as those varieties and organs with the highest accumulations, as well as the optimal age and growth conditions for each variety. The highest amounts of cichoric acid were measured from the older, wild inflorescences of E. pallida var.sanguinea, whereas the highest quantities of the alkamides 1-3 and 7 were present in roots of wild and transplanted E. purpurea. Baseline phytochemical data and chromatographic profiles for all types of wild Echinacea may be used for protection of wild stands, germplasm identification, and crop improvement.     

Supercritical fluid extraction of alkylamides from Echinacea angustifolia

Echinacea has been known for its immunostimulatory activity, and its alkylamide components have been linked to such biological activity. Consequently, alkylamides in Echinacea angustifolia were extracted using supercritical carbon dioxide from fresh and dried roots at 45-60 degrees C and 34-55 MPa, and the alkylamide yield in the extracts was determined. The yield of alkylamides from fresh roots increased with temperature yet decreased with pressure, whereas the yield from air-dried roots (moisture content 8.4%) increased with both temperature and pressure. Freeze-drying of the roots to a moisture content of 4.9% did not result in any further increase in the yield compared to that of air-dried roots. Alkylamide yield of the ground dried roots extract was the highest (p < or = 0.05) among those from fresh, ground and unground E. angustifolia roots. Supercritical fluid extraction therefore shows potential for the recovery of alkylamides from dried Echinacea roots.     

Pomegranate phenolics from the peels, arils, and flowers are antiatherogenic: studies in vivo in atherosclerotic apolipoprotein e-deficient (E 0) mice and in vitro in cultured macrophages and lipoproteins

We have analyzed in vivo and in vitro the antiatherogenic properties and mechanisms of action of all pomegranate fruit parts: peels (POMxl, POMxp), arils (POMa), seeds (POMo), and flowers (POMf), in comparison to whole fruit juice (PJ). Atherosclerotic E 0 mice consumed POM extracts [200 microg of gallic acid equivalents (GAE)/mouse/day] for 3 months. Blood samples, peritoneal macrophages (MPM), and aortas were then collected. All POM extracts possess antioxidative properties in vitro. After consumption of PJ, POMxl, POMxp, POMa, or POMf by E (0) mice, the atherosclerotic lesion area was significantly decreased by 44, 38, 39, 6, or 70%, respectively, as compared to placebo-treated group, while POMo had no effect. POMf consumption reduced serum lipids, and glucose levels by 18-25%. PJ, POMxl, POMxp, POMf, or POMa consumption resulted in a significant decrement, by 53, 42, 35, 27, or 13%, respectively, in MPM total peroxides content, and increased cellular paraoxonase 2 (PON2) activity, as compared to placebo-treated mice. The uptake rates of oxidized-LDL by E (0)-MPM were significantly reduced by approximately 15% after consumption of PJ, POMxl, or POMxp. Similar results were obtained on using J774A.1 macrophage cell line. Finally, pomegranate phenolics (punicalagin, punicalin, gallic acid, and ellagic acid), as well as pomegranate unique complexed sugars, could mimic the antiatherogenic effects of pomegranate extracts. We conclude that attenuation of atherosclerosis development by some of the POM extracts and, in particular, POMf, could be related to the combined beneficial effects on serum lipids levels and on macrophage atherogenic properties.     

Analysis of alkylamides in Echinacea plant materials and dietary supplements by ultrafast liquid chromatography with diode array and mass spectrometric detection

Alkylamides are a class of compounds present in plants of the genus Echinacea (Asteraceae), which have been shown to have high bioavailability and immunomodulatory effects. Fast analysis to identify these components in a variety of products is essential to profile products used in clinical trials and for quality control of these products. A method based on ultrafast liquid chromatography (UFLC) coupled with diode array detection and electrospray ionization mass spectrometry was developed for the analysis of alkylamides from the roots of Echinacea angustifolia (DC.) Hell., Echinacea purpurea (L.) Moench, and commercial dietary supplements. A total of 24 alkylamides were identified by LC-MS. The analysis time for these components is 15 min. Compared to the alkylamide profiles determined in the Echinacea root materials, the commercial products showed a more complex profile due to the blending of root and aerial parts of E. purpurea. This versatile method allows for the identification of alkylamides in a variety of Echinacea products and presents the most extensive characterization of alkylamides in E. angustifolia roots so far.     

Characterization and functional study of Antrodia camphorata lipopolysaccharide

Lipopolysaccharide (LPS) is a highly proinflammatory molecule isolated from bacteria. This study demonstrated the existence of LPS in a medicinal fungus, Antrodia camphorata. Because no LPS had been identified in any fungus organism, the purification of LPS from A. camphorata was attempted. LPSs from six strains of A. camphorata (35396, 35398, 35716, B71, B85, and B86) were isolated. Chemical and functional properties were investigated on the fungus LPS. Compositional analysis revealed that sorbitol, fucose, galactose, and glucose were the neutral sugars in LPS of A. camphorata. Galactosamine, glucosamine, galactose, and glucose were the predominant monosaccharide species in E. coli O129 LPS molecules, whereas galactosamine and glucosamine were absent in A. camphorata LPS. Because these properties are different from those of bacterial LPS, the functions between fungus and bacterial LPS are also discussed. The vascular endothelial lining of blood vessels, which controls leucocyte traffic and activation, may be one of the primary targets of LPS action during sepsis. Assays for biological activity were performed on endothelial cells with anti-inflammatory effects associated with sepsis. A. camphorata LPS apparently showed a lesser extent of cytotoxicity than bacterial LPS. In contrary to the proinflammatory property of bacterial LPS, LPS from A. camphorata differentially reversed bacterial LPS-induced intercellular adhersion molecule-1 and monocyte adhesion; both were indicators during inflammatory process. In conclusion, basic chemical properties categorized A. camphorata extracts into lipopolysaccharide. However, the detailed functional structures and bioactivities of A. camphorata LPS were totally different from those of bacterial LPS. The investigation of the existence and anti-inflammatory effect of fungus LPS is at present a truly novel and important finding. These results show that LPS isolated from A. camphorata offers a novel therapeutic target for anti-inflammation against E. coli infection.     

Hericium erinaceus mushroom extracts protect infected mice against Salmonella Typhimurium-Induced liver damage and mortality by stimulation of innate immune cells

The present study investigated the antibacterial effect of four extracts from the fruitbody of the edible medicinal mushroom Hericium erinaceus (hot water extract, HWE; microwave/50% ethanol extract, MWE; acid extract, ACE; and alkaline extract, AKE) against murine salmonellosis. The extracts had no effect on Salmonella ser. Typhimurium growth in culture. Nor were the extracts toxic to murine macrophage cells, RAW 264.7. HWE and MWE stimulated uptake of the bacteria into the macrophage cells as indicated by increased colony-forming unit (CFU) counts of the contents of the lysed macrophages infected with Salmonella Typhimurium for 30 and 60 min. Two hours postinfection, the bacterial counts increased in the macrophages, but 4 and 8 h postinfection the HWE- and MWE-treated cells showed greater activity against the bacteria than the control. HWE- and MWE-treated noninfected macrophages had altered morphology and elevated inducible nitric oxide (NO) synthase (iNOS) mRNA expression. In the presence of S. Typhimurium, iNOS mRNA expression was further increased, accompanied by an increase in NO production. Histology assays of the livers of mice infected with a sublethal dose (1 × 10(4) CFU) of S. Typhimurium showed that HWE and MWE, administered by daily intraperitoneal injection, protected against necrosis of the liver, a biomarker of in vivo salmonellosis. The lifespans of mice similarly infected with a lethal dose of S. Typhimurium (1 × 10(5) CFU) were significantly extended by HWE and MWE. β-Glucan, known to stimulate the immune system, was previously found to be present in high amounts in the active extracts. These results suggest that the mushroom extract activities against bacterial infection in mice occur through the activation of innate immune cells.     

Partial characterization of polyphenol oxidase activity in raspberry fruits.

A partial characterization of polyphenol oxidase (PPO) activity in raspberry fruits is described. Two early cultivars harvested in May/June (Heritage and Autumm Bliss) and two late cultivars harvested in October-November (Ceva and Rubi) were analyzed for PPO activity. Stable and highly active PPO extracts were obtained using insoluble poly(vinylpyrrolidone) (PVP) and Triton X-100 in sodium phosphate, pH 7.0 buffer. Polyacrylamide gel electrophoresis of raspberry extracts under nondenaturing conditions resolved in one band (R(f)()(1) = 0.25). Raspberry PPO activity has pH optima of 8.0 and 5.5, both with catechol (0.1 M). Maximum activity was with D-catechin (catecholase activity), followed by p-coumaric acid (cresolase activity). Heritage raspberry also showed PPO activity toward 4-methylcatechol. Ceva and Autumm Bliss raspberries showed the higher PPO activity using catechol as substrate.     

Total phenolics and antioxidant activities of fenugreek, green tea, black tea, grape seed, ginger, rosemary, gotu kola, and ginkgo extracts, vitamin E, and tert-butylhydroquinone

The total phenolics and antioxidant activities of fenugreek, green tea, black tea, grape seed, ginger, rosemary, gotu kola, and ginkgo extracts, vitamin E, and tert-butylhydroquinone, were determined. Grape seed and green tea were analyzed for their phenolic constituents using high-performance liquid chromatography. The total phenolics of the plant extracts, determined by the Folin-Ciocalteu method, ranged from 24.8 to 92.5 mg of chlorogenic acid equivalent/g dry material. The antioxidant activities of methanolic extracts determined by conjugated diene measurement of methyl linoleate were 3.4-86.3%. The antioxidant activity of the extracts using chicken fat by an oxidative stability instrument (4.6-10.2 h of induction time) followed a similar trend in antioxidant activity as determined by the Folin-Ciocalteu method. Seven phenolics in grape seed and green tea extracts were identified that ranged from 15.38 to 1158.49 and 18.3 to 1087.02 mg/100 g of extract, respectively. Plant extracts such as green tea and grape seed extracts can be used to retard lipid oxidation in a variety of food products.     

Chromatographic methods for metabolite profiling of virus- and phytoplasma-infected plants of Echinacea purpurea

This study was focused on the effects of virus and phytoplasma infections on the production of Echinacea purpurea (L.) Moench secondary metabolites, such as caffeic acid derivatives, alkamides, and essential oil. The identification of caffeic acid derivatives and alkamides was carried out by means of high-performance liquid chromatography-diode array detection (HPLC-DAD), HPLC-electrospray ionization-mass spectrometry (ESI-MS), and MS(2). Quantitative analysis of these compounds was carried out using HPLC-DAD. The results indicated that the presence of the two pathogens significantly decreases (P < 0.05) the content of cichoric acid, the main caffeic acid derivative. Regarding the main alkamide, dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamide, a significant decrease (P < 0.05) in the content of this secondary metabolite was observed in virus-infected plants in comparison with healthy plants, while in the phytoplasma-infected sample the variation of this secondary metabolite was not appreciable. The % relative area of the E/Z isomers of this alkamide was also found to change in infected samples. The gas chromatography (GC) and GC-MS analysis of E. purpurea essential oil enabled the identification of 30 compounds. The main significant differences (P < 0.05) in the semiquantitative composition were observed for three components: limonene, cis-verbenol, and verbenone. The results indicate that the presence of virus and phytoplasma has an appreciable influence on the content of E. purpurea secondary metabolites, which is an important issue in defining the commercial quality, market value, and therapeutic efficacy of this herbal drug.     

Antioxidant activities and phenolic composition of extracts from Greek oregano,Greek sage,and summer savory

Oregano vulgare L. ssp. hirtum (Greek oregano), Salvia fruticosa (Greek sage), and Satureja hortensis (summer savory) were examined as potential sources of phenolic antioxidant compounds. The antioxidant capacities (antiradical activity by DPPH* test, phosphatidylcholine liposome oxidation, Rancimat test) and total phenol content were determined in the ethanol and acetone extracts of the dried material obtained from the botanically characterized plants. The ground material was also tested by the Rancimat test for its effect on the stability of sunflower oil. The data indicated that ground material and both ethanol and acetone extracts had antioxidant activity. Chromatographic (TLC, RP-HPLC) and NMR procedures were employed to cross-validate the presence of antioxidants in ethanol and acetone extracts. The major component of all ethanol extracts was rosmarinic acid as determined by RP-HPLC and NMR. Chromatography indicated the presence of other phenolic antioxidants, mainly found in the acetone extracts. The presence of the flavones luteolin and apigenin and the flavonol quercetin was confirmed in the majority of the extracts by the use of a novel (1)H NMR procedure, which is based on the strongly deshielded OH protons in the region of 12-13 ppm without previous chromatographic separation. This deshielding may be attributed to the strong intramolecular six-membered ring hydrogen bond of the OH(5)...CO(4) moiety.     

THE MOBILIZATION OF IRON BY EXTRACTS OF EUCALYPTUS LEAF LITTER

When aqueous extracts of leaf litter from four closely related Eucalyptus species were reacted with soil material under aerobic conditions their iron mobilizing activity was found to be inversely related to the productivity of the sites on which the species grew. The activity of litter extracts of the four species grown in similar soils in the field was found to increase in the order E. regnans, E. obliqua, E. radiata, E. sieberiana. The results indicate that species-soil interactions could be as important as inherent species characteristics in determining whether or not a species is a ‘podzol former’. The effect on the activity of the extracts of altering their pH before reaction with either soil or with prepared iron oxides suggests that, whereas organic acids could be mainly responsible for mobilizing iron from soil and from anhydrous ferric oxide, polyphenols could be more important than organic acids in mobilizing iron from hydrous ferric oxide. The pH and E_h curves obtained when litter extracts were titrated with a ferric chloride solution showed that iron from this source was strongly reduced by the extracts at low pH.     

Antioxidant capacity of different broccoli (Brassica oleracea) genotypes using the oxygen radical absorbance capacity (ORAC) assay

Antioxidant capacity of hydrophilic and lipophilic extracts from eight broccoli genotypes was compared using the oxygen radical absorbance capacity (ORAC) assay. Each genotype was analyzed for carotenoid, tocopherol, ascorbic acid, and flavonoid content. Results indicate that the antioxidant capacity of hydrophilic extracts ranged from 65.8 to 121.6 micromol trolox equivalents (TE)/g of tissue, and the capacity of lipophilic extracts ranged from 3.9 to 17.5 micromol TE/g. Ascorbic acid and flavonoid content of the hydrophilic extracts did not explain the total variation in antioxidant capacity of those extracts, suggesting either the presence of other antioxidant components that have yet to be identified or that the known antioxidants are producing synergistic effects. The carotenoids did correlate with antioxidant capacity of the lipophilic extracts and accounted for the majority of the variability in that fraction. The variability in hydrophilic and lipophilic antioxidant capacity found among these genotypes suggests that potential efficacy from antioxidants will vary considerably from genotype to genotype.     

Effect of selenium on increasing the antioxidant activity of tea leaves harvested during the early spring tea producing season

This research was to determine the effect of foliar application of selenium on increasing the antioxidant activity of tea harvested during the early spring tea producing season using a alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radical scavenging method and the linoleic acid system. The results showed that the radical scavenging ability of the tea extracts followed this order during the first 60 min: selenium-enriched tea obtained by fertilization with selenate > BHT > selenium-enriched tea obtained by fertilization with selenite > alpha-tocopherol > regular tea. Se-enriched tea obtained by fertilization with selenate exhibited the highest inhibition percentage of 84.29% at 30 min. Se-enriched tea extracts provided higher hydrogen-donating capabilities than regular tea and contrasts with BHT and alpha-tocopherol at the concentration of 100 microg of solids/mL of ethanol. There was a little change in the sequence of radical scavenging ability during the later 60 min: Se-enriched tea obtained by fertilization with selenate > Se-enriched tea obtained by fertilization with selenite > BHT > regular tea > alpha-tocopherol. The individual activity of tea extracts and references measured by the linoleic acid system showed that the tea extracts, BHT, and alpha-tocopherol manifested almost the same patterns of activity as the DPPH method. Tea enriched in selenium by fertilization with selenate still exhibited the highest inhibition activity of lipid oxidation, whereas alpha-tocopherol showed the lowest inhibition. The antioxidant activity of Se-enriched green tea harvested during the early spring tea producing season is enhanced compared to regular tea.     

Impact of cultivar, harvesting time, and seasonal variation on the content of biophenols in olive mill waste

Olive mill waste (OMW) contains substantial amounts of valuable antioxidant biophenols that can be recovered for possible applications in food, pharmaceutical, and cosmetic industries. However, the impact of cultivar, harvesting time, and seasonal variation on the phenolic composition of OMW has not yet been assessed. Total phenols, antioxidant activity, and phenol profiles of OMW extracts from five different Australian-grown cultivars (Barnea, Correggiola, Manzanillo, Mission, and Paragon) were studied at four different harvesting times in the 2004 season. The impact of seasonal variation was assessed by comparing total phenol content, antioxidant activity, and phenol profile of two cultivars (Correggiola and Mission) harvested in the 2004 and 2005 seasons. The phenol content and antioxidant activity at different harvesting times were mainly a function of the olive cultivar. Harvesting time had a quantitative effect rather than a qualitative effect on the phenol profile. Intercultivar and harvesting time variation accounted for a 2-5-fold change in the total phenol and antioxidant capacity, while levels of individual biophenols experienced up to 50-fold change. The phenol content and antioxidant capacity of OMW significantly changed between seasons with different variation patterns for different cultivars.     

Enhanced anti-inflammatory activities of Monascus pilosus fermented products by addition of ginger to the medium

Hypercholesterolemia initiates the atherogenic process; however, chronic inflammation promotes atherogenesis. Monascus spp. fermented products are recognized for their anti-hypercholesterolemic effect, but their anti-inflammatory activity is not as significant as that of many plant-derived foods. To enhance the anti-inflammatory function of Monascus pilosus fermented products, ginger was added to the PDB medium at a ratio of 20% (v/v). The mycelia and broth were collected, freeze-dried, and extracted by ethanol for assays. Macrophage RAW264.7 was challenged with lipopolysaccharide (LPS) and coincubated with the extracts of fermented product cultured in ginger-supplemented medium (MPG) or extracts of fermented product cultured in regular PDB medium (MP) for 18 h. Human umbilical vein endothelial cell HUVEC was challenged with tumor necrosis factor (TNF)-α and coincubated with the extracts of either MPG or MP for 6 h. The results showed that MPG significantly (p<0.05) lowered the production of macrophage pro-inflammatory cytokines TNF-α, nitric oxide (NO), interleukin (IL)-1, IL-6, and prostaglandin E2 (PGE2) by 68.53%, 84.29%, 32.55%, 84.49%, and 69.49%, respectively; however, MP had no inhibitory effect. MPG significantly downregulated the expression of p-IκB, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) in macrophage by 42.16%, 50.87%, and 51.35%, respectively, while MP had no inhibition on COX-2 expression and only 16.64% and 19.22% downregulatory effect on iNOS and phosphorylated-IκB (p-IκB), respectively. Moreover, MPG significantly suppressed the expression of vessel cell adhesion molecule-1 (VCAM-1) and p-IκB in endothelial cell by 63.48% and 63.41%, respectively. LC/MS/MS analysis indicated that 6-gingerdiol was formed in the ginger-modified medium during fermentation. The results of this study will facilitate the development of Monascus spp. fermented products as antiatherosclerotic nutraceuticals.