Chemical Compositions of Scutellaria baicalensis Georgi. (Huangqin) Extracts and Their Effects on ACE2 Binding of SARS-CoV-2 Spike Protein, ACE2 Activity, and Free Radicals.

The water and ethanol extracts of huangqin, the roots of Scutellaria baicalensis Georgi. with potential antiviral properties and antioxidant activities, were investigated for their chemical profiles and their abilities to interfere with the interaction between SARS-CoV-2 spike protein and ACE2, inhibiting ACE2 activity and scavenging free radicals. A total of 76 compounds were tentatively identified from the extracts. The water extract showed a greater inhibition on the interaction between SARS-CoV-2 spike protein and ACE2, but less inhibition on ACE2 activity than that of the ethanol extract on a per botanical weight concentration basis. The total phenolic content was 65.27 mg gallic acid equivalent (GAE)/g dry botanical and the scavenging capacities against HO●, DPPH●, and ABTS●+ were 1369.39, 334.37, and 533.66 µmol trolox equivalent (TE)/g dry botanical for the water extract, respectively. These values were greater than those of the ethanol extract, with a TPC of 20.34 mg GAE/g, and 217.17, 10.93, and 50.21 µmol TE/g against HO●, DPPH●, and ABTS●+, respectively. The results suggested the potential use of huangqin as a functional food ingredient in preventing COVID-19.

Bioactive Compositions of Cinnamon (Cinnamomum verum J. Presl) Extracts and Their Capacities in Suppressing SARS-CoV-2 Spike Protein Binding to ACE2, Inhibiting ACE2, and Scavenging Free Radicals.

Cinnamon (Cinnamomum verum J. Presl) bark and its extracts are popular ingredients added to food and supplement products. It has various health effects, including potentially reducing the risk of coronavirus disease-2019 (COVID-19). In our study, the bioactives in cinnamon water and ethanol extracts were chemically identified, and their potential in suppressing SARS-CoV-2 spike protein-angiotensin-converting enzyme 2 (ACE2) binding, reducing ACE2 availability, and scavenging free radicals was investigated. Twenty-seven and twenty-three compounds were tentatively identified in cinnamon water and ethanol extracts, respectively. Seven compounds, including saccharumoside C, two emodin-glucuronide isomers, two physcion-glucuronide isomers, and two type-A proanthocyanidin hexamers, were first reported in cinnamon. Cinnamon water and ethanol extracts suppressed the binding of SARS-CoV-2 spike protein to ACE2 and inhibited ACE2 activity in a dose-dependent manner. Cinnamon ethanol extract had total phenolic content of 36.67 mg gallic acid equivalents (GAE)/g and free radical scavenging activities against HO• and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation (ABTS•+) of 1688.85 and 882.88 µmol Trolox equivalents (TE)/g, which were significantly higher than those of the water extract at 24.12 mg GAE/g and 583.12 and 210.36 µmol TE/g. The free radical scavenging activity against 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) of cinnamon ethanol extract was lower than that of the water extract. The present study provides new evidence that cinnamon reduces the risk of SARS-CoV-2 infection and COVID-19 development.

Quantification of cranberry proanthocyanidins by normal-phase high-performance liquid chromatography using relative response factors.

INTRODUCTION: American cranberries (Vaccinium macrocarpon) contain primarily A-type proanthocyanidins (PACs), which have been shown to prevent urinary tract infection. Currently, the accurate quantification of cranberry PACs is still lacking. OBJECTIVE: A normal-phase high-performance liquid chromatography (NP-HPLC) method using relative response factors was developed and validated to quantify cranberry PAC oligomers and polymers. MATERIALS AND METHODS: PAC oligomers with degree of polymerisation (DP) 3-9 and total polymers were isolated from the cranberry juice concentrate. Characterisation of the isolated PAC oligomers was performed by ultra-performance liquid chromatography-high resolution mass spectrometry. The relative response factors of oligomers from DP 2-9 and total polymers were determined against procyanidin A2. Method validation was conducted to assess limit of detection, limit of quantification, the linearity and working range, precision and accuracy. In addition, quantifications of PACs by NP-HPLC using relative response factors and two other commonly used methods were conducted in three cranberry food products. RESULTS: Cranberries PACs oligomers contained both A-type and B-type linkage, with epicatechin and epigallocatechin as basic units. Method validation results suggested this method is reliable and reproducible. Quantifications of PACs by NP-HPLC using relative response factors yielded higher values than that by the other two methods. CONCLUSION: A NP-HPLC method using the relative response factors was developed and validated. This method provides a more accurate approach in determining cranberry PACs. It can be used to quantify individual oligomers from DP 2-9, total polymers and total PACs in cranberries and cranberry products.

Analysis of cranberry proanthocyanidins using UPLC-ion mobility-high-resolution mass spectrometry.

Cranberry proanthocyanidin oligomers were investigated using ultra performance liquid chromatography-ion mobility-high-resolution mass spectrometry (UPLC-IM-HRMS). A total of 304 individual A-type and B-type proanthocyanidins, including 40 trimers, 68 tetramers, 53 pentamers, 54 hexamers, 49 heptamers, 28 octamers, and 12 nonamers, were characterized. A-type proanthocyanidins appeared to dominate the cranberry proanthocyanidins. As the degree of polymerization increased, the abundance of molecules with multiple A-type double inter-flavan linkage or having doubly charged ions also increased. Under the same charge state, the drift times of proanthocyanidin ions increased with their degree of polymerization or the number of double inter-flavan linkages. For the same proanthocyanidin molecules, doubly charged ions had shorter drift times compared to their singly charged counterparts, which lead to separated trendlines in the ion mobility-mass plot. While consistent ion mobility was observed for most proanthocyanidins with the same degree of polymerization, coeluted isomeric ions of trimer and tetramer were detected by their unique drift times. Incorporation of ion mobility into HRMS proved to be of great value to characterize and analyze proanthocyanidins from complex sample matrices. Graphical abstract.

Prevention of Atherosclerosis by Berries: The Case of Blueberries.

Berry consumption has been associated with cardiovascular disease prevention in recent years. Atherosclerosis is one of the major causes of cardiovascular diseases. However, research on the prevention of atherosclerosis through consuming individual whole berries, specifically direct evidence, remains scarce. Therefore, further elucidating the role that berries play in the prevention of atherosclerosis is warranted. In this perspective, blueberries were selected to articulate research strategies for studying atheroprotective effects of berries. Studies from human subjects and various animal models are summarized. The mechanisms by which blueberries may act, through reducing oxidative stress, decreasing inflammation, improving endothelial dysfunction, regulating cholesterol accumulation and trafficking, along with potentially influencing gut microbiota, are also discussed. Blueberries contain high levels of polyphenolic compounds, which were widely indicated as major bioactive compounds. Nonetheless, the metabolites/catabolites after blueberry consumption, such as simple phenolic acids, rather than original compounds in berries, may be the actual in vivo bioactive compounds. Future research should focus on obtaining more direct evidence, preferably in humans, understanding of the mechanisms of action at the molecular level, and identifying bioactive compounds as well as which compounds act synergistically to convey health benefits. The research strategy discussed here may also be applied to the studies of other fruits and berries.

(-)-Epicatechin regulates blood lipids and attenuates hepatic steatosis in rats fed high-fat diet.

SCOPE: (-)-Epicatechin (EC) is a natural flavanol monomer found in cocoa, green tea, and a variety of other plant foods. In this study, effects of EC on blood lipids and hepatic steatosis, and the underlying mechanisms were investigated. METHODS AND RESULTS: A hyperlipidemic rat model was induced by high-fat, high-cholesterol diet. EC was then administrated to the animals by gavage at doses of 10, 20, 40 mg/kg body weight (BW) for 12 weeks. Simvastatin was included as a positive control. The results showed that EC significantly reduced total cholesterol, LDL cholesterol and triglyceride, alleviated liver fat accumulation, while increased HDL cholesterol, in hyperlipidemic rats. EC also reduced lipid peroxidation, inhibited the pro-inflammatory cytokines, and lowered serum AST and ALT. The potential molecular mechanisms of EC underlying these effects were proposed to be associated to regulating Insig-1-SREBP-SCAP pathway, and other lipid metabolic related genes including LXR-α, FAS, and SIRT1. CONCLUSION: EC effectively improved blood lipid profile and protected liver from accumulating excessive fat in hyperlipidemic rats. The results shed a light on the potential role of EC as a promising natural product in preventing hyperlipidemia and nonalcoholic fatty liver disease.

Differential effects of short term feeding of a soy protein isolate diet and estrogen treatment on bone in the pre-pubertal rat.

BACKGROUND: Previous reports suggest that beneficial effects of soy on bone quality are due to the estrogenic actions of isoflavone phytochemicals associated with the protein. However, mechanistic studies comparing the effects of soy diet and estrogens on bone, particularly in rapidly growing animals are lacking. METHODOLOGY AND PRINCIPAL FINDINGS: We studied the effects of short term feeding of soy protein isolate (SPI) on bone in comparison to the effects of 17ß-estradiol (E2) in pre-pubertal rats. Female rats were weaned to one of 4 treatments: 1) a control casein-based diet (CAS); 2) CAS with subcutaneous E2 (10 µg/kg/d) (CAS+E2); 3) a SPI-containing diet (SPI); or 4) SPI with subcutaneous E2 (SPI) or SPI with 10 µg/kg/d E2 (SPI+E2) for 14 days beginning on postnatal day 20. SPI increased while E2 decreased bone turnover compared to CAS. In contrast, both treatments decreased serum sclerostin levels. Microarray analysis of RNA isolated from bone revealed 652 genes regulated by SPI, 491 genes regulated by E2, and 266 genes regulated by both SPI diet and E2 compared to CAS. The expression of caveolin-1, a protein localized in the cell membrane, was down-regulated (p<0.05) in rats fed SPI, but not by E2 compared to rats fed casein. Down-regulation of caveolin-1 by SPI was associated with increased BMP2, Smad and Runx2 expression in bone and osteoblasts (p<0.05). CONCLUSIONS/SIGNIFICANCE: These results suggest SPI and E2 have different effects on bone turnover prior to puberty. Approximately half of the genes are regulated in the same direction by E2 or SPI, but in combination, SPI blocks the estrogen effects and returns the profile towards control levels. In addition, there are E2 specific and SPI-specific gene changes related to regulation of bone formation.

Repression of mammosphere formation of human breast cancer cells by soy isoflavone genistein and blueberry polyphenolic acids suggests diet-mediated targeting of cancer stem-like/progenitor cells.

Mammary stem cells are undifferentiated epithelial cells, which initiate mammary tumors and render them resistant to anticancer therapies, when deregulated. Diets rich in fruits and vegetables are implicated in breast cancer risk reduction, yet underlying mechanisms are poorly understood. Here, we addressed whether dietary factors selectively target mammary epithelial cells that display stem-like/progenitor subpopulations with previously recognized tumor-initiating potential. Using estrogen receptor-positive MCF-7 and estrogen receptor-negative MDA-MB-231 human breast cancer cell lines and freshly isolated epithelial cells from MMTV-Wnt-1 transgenic mouse mammary tumors, we demonstrate that sera of adult mice consuming soy isoflavone genistein (GEN) or blueberry (BB) polyphenol-containing diets alter the population of stem-like/progenitor cells, as measured by their functional ability to self-renew and form anchorage-independent spheroid cultures in vitro at low frequency (1-2%). Serum effects on mammosphere formation were dose-dependently replicated by GEN (40 nM >2 µM) and targeted the basal stem-like CD44+/CD24-/ESA+ and the luminal progenitor CD24+ subpopulations in MDA-MB-231 and MCF-7 cells. GEN inhibition of mammosphere formation was mimicked by the Akt inhibitor perifosine and was associated with enhanced tumor suppressor phosphatase and tensin homologue deleted on chromosome ten (PTEN) expression. In contrast, a selected mixture of BB phenolic acids was only active in MDA-MD-231 cells and its CD44+/CD24-/ESA+ subpopulation, and this activity was independent of induction of PTEN expression. These findings delineate a novel and selective function of distinct dietary factors in targeting stem/progenitor cell populations in estrogen receptor-dependent and -independent breast cancers.

Lowbush blueberries inhibit scavenger receptors CD36 and SR-A expression and attenuate foam cell formation in ApoE-deficient mice.

Blueberries have recently been reported to reduce atherosclerotic lesion progression in apoE deficient (apoE(-/-)) mice. However, the underlying mechanisms are not fully understood. The objective of this study was to determine whether lowbush blueberries altered scavenger receptor expression and foam cell formation in apoE(-/-) mice. ApoE(-/-) mice were fed AIN-93 diet (CD) or CD formulated to contain 1% freeze-dried lowbush blueberries (BB) for 20 weeks. Gene expression and protein levels of scavenger receptor CD36 and SR-A in aorta and thioglycollate-elicited peritoneal macrophages (PM) were lower in mice fed BB (P < 0.05). In the second experiment, apoE(-/-) mice were fed CD or BB for 5 weeks. PM were collected and cultured. Gene expression and protein levels of CD36 and SR-A were found to be lower in PM of BB fed mice (P < 0.05). In PM from BB fed mice, fewer oxLDL-induced foam cells were formed compared to those from mice fed CD. Gene expression and protein levels of PPARγ were lower in the PM of BB fed mice (P < 0.05). Detectable isomers of hydroxyoctadecadienoic acids (HODEs) and hydroxyeicosatetraenoic acid (HETEs) were also lower in the PM of BB fed mice (P < 0.05 or P < 0.01). In conclusion, BB inhibited expression of the two major scavenger receptors CD36 and SR-A in PM of apoE(-/-) mice, at least in part through down-regulating PPARγ and reducing its endogenous ligands HODEs and HETEs. We proposed that BB mediated reduction of scavenger receptor expression and attenuation of oxLDL-induced foam cell formation in PM of apoE(-/-) mice are important mechanisms of the athero-protective effects of BB.

Biflavonoids from Caper (Capparis spinosa L.) fruits and their effects in inhibiting NF-kappa B activation.

Caper (Capparis spinosa L.) fruits have been widely used as food and folk medicine in the Mediterranean basin and in central and west Asia. In this study, two biflavonoids, isoginkgetin, and ginkgetin, together with three other flavonoids, were isolated from caper fruits. Their chemical structures were elucidated by spectroscopic analyses and comparison with literature. To our knowledge, isoginkgetin, ginkgetin and sakuranetin were identified in caper for the first time. Notably, it is also the first time that biflavonoids have ever been found in the Capparidaceae. Concentrations of the two biflavonoids were measured in caper fruits collected from four major growing areas in northwest China. The anti-inflammatory effects of the flavonoids from caper fruits were evaluated by secreted placental alkaline phosphatase (SEAP) reporter assay, which was designed to measure nuclear factor-kappa B (NF-κB) activation. Isoginkgetin and ginkgetin showed inhibitory effects in initial screen at 20 µM, while the effect of ginkgetin was much greater than that of isoginkgetin. In a dose-response experiment, the IC(50) value of ginkgetin was estimated at 7.5 µM, suggesting it could be a strong NF-κB inhibitor and worthy of study in vivo.

Anti-inflammatory effects of caper (Capparis spinosa L.) fruit aqueous extract and the isolation of main phytochemicals.

Caper (Capparis spinosa L.) fruits have been used as food as well as folk medicine in the treatment of inflammatory disorders, such as rheumatism. The present study was carried out to study the anti-inflammatory activities of C. spinosa L. fruit (CSF) aqueous extract and to isolate main phytochemicals from its bioactive fractions. The CSF aqueous extract were separated into three fractions (CSF1-CSF3) by macroporous adsorption resins. The fractions CSF2 and CSF3 effectively inhibited the carrageenan-induced paw edema in mice. Systematic fractionation and isolation from CSF2+3 led to the identification of 13 compounds (1-13). Their chemical structures were elucidated by spectroscopic analyses including nuclear magnetic resonance (NMR) and mass spectrometry (MS) and literature comparisons. Major compounds found in the bioactive fraction CSF2+3 are flavonoids, indoles, and phenolic acids. To our knowledge, 8 of these 13 compounds (1-4, 6-7, 10, and 13) were identified from caper fruits for the first time. The anti-inflammatory effects of these purified compounds are currently under investigation.

Non-isoflavone phytochemicals in soy and their health effects.

Epidemiological and clinical studies have linked consumption of soy foods with low incidences of a number of chronic diseases, such as cardiovascular diseases, cancer, and osteoporosis. Over the past decades, enormous research efforts have been made to identify bioactive components in soy. Isoflavones and soy protein have been suggested as the major bioactive components in soy and have received considerable attention. However, there are hundreds of phytochemical components in soybeans and soy-based foods. In recent years, accumulating evidence has suggested that the isoflavones or soy proteins stripped of phytochemicals only reflect certain aspects of health effects associated with soy consumption. Other phytochemicals, either alone or in combination with isoflavones or soy protein, may be involved in the health effects of soy. This review attempts to summarize major non-isoflavone phytochemicals in soy, as well as their bioavailability and health effects. In addition, a brief discussion of components formed during food processing is also included.

Dietary black raspberry anthocyanins do not alter development of obesity in mice fed an obesogenic high-fat diet.

Anthocyanins (ACNs) from various foods have been shown to minimize the development of obesity in some animal models. The objective of the current study was to compare the effects of feeding purified black raspberry (BRB) ACNs or the freeze-dried whole BRB on the development of obesity. Male C57BL/6J mice (25 days of age) were assigned at random to treatments (7/treatment; 3/cage). The treatments included (1) control low-fat diet (10% calories from fat) (LF); (2) LF plus BRB juice in place of drinking water; (3) LF diet plus purified BRB ACNs in drinking water (1.25 mg/mL); (4) control high-fat diet (60% calories from fat) (HF60); (5) HF60 diet + BRB juice in place of drinking water; (6) HF60 diet + ACNs in drinking water (1.25 mg/mL); and (7) HF60 + freeze-dried whole BRB powder (21.7 g/kg of diet). Body weight gains in mice fed HF60 diet plus purified BRB ACNs tended to be lower after 56, 63, and 70 days than in mice fed HF60 alone. Body weights were increased at time of sacrifice, but heart, liver, and kidney weights as a percentage of body weight were decreased in mice fed HF60 diet compared to LF fed mice. Weights (g or g/body weight) of epididymal and retroperitoneal fat were increased in the HF60 fed mice compared to LF fed mice. Fasting serum glucose, leptin, and insulin levels as well as homeostasis assessment of insulin resistance (HOMA-IR) were elevated in mice fed the HF60 diet relative to LF-fed controls. Serum cholesterol, triglycerides, and monocyte chemoattractant protein-1 (MCP-1) were not altered by diet. Serum levels of resistin were increased in mice fed the HF60 diet compared to mice fed the LF diet. None of the responses measured were altered by whole BRB powder included in the diet relative to the HF60 control diet. Cyanidin containing di- or triglycosides in BRB was ineffective in altering the development of obesity in contrast to cyanidin-monoglycosides, which have been shown to be effective. The sugar moiety on the anthocyanidins may be an important factor in determining the response in the development of obesity.

Purified blueberry anthocyanins and blueberry juice alter development of obesity in mice fed an obesogenic high-fat diet.

Male C57BL/6J mice (25 days of age) were fed either a low-fat diet (10% kcal from fat) (LF) or a high-fat diet (45% kcal from fat) (HF45) for a period of 72 days. Blueberry juice or purified blueberry anthocyanins (0.2 or 1.0 mg/mL) in the drinking water were included in LF or HF45 treatments. Sucrose was added to the drinking water of one treatment to test if the sugars in blueberry juice would affect development of obesity. Total body weights (g) and body fat (%) were higher and body lean tissue (%) was lower in the HF45 fed mice compared to the LF fed mice after 72 days, but in mice fed HF45 diet plus blueberry juice or blueberry anthocyanins (0.2 mg/mL), body fat (%) was not different from those mice fed the LF diet. Anthocyanins (ACNs) decreased retroperitoneal and epididymal adipose tissue weights. Fasting serum glucose concentrations were higher in mice fed the HF45 diet. However, it was reduced to LF levels in mice fed the HF45 diet plus 0.2 mg of ACNs/mL in the drinking water, but not with blueberry juice. beta cell function (HOMA-BCF) score was lowered with HF45 feeding but returned to normal levels in mice fed the HF45 diet plus purified ACNs (0.2 mg/mL). Serum leptin was elevated in mice fed HF45 diet, and feeding either blueberry juice or purified ACNs (0.2 mg/mL) decreased serum leptin levels relative to HF45 control. Sucrose in drinking water, when consumption was restricted to the volume of juice consumed, produced lower serum leptin and insulin levels, leptin/fat, and retroperitoneal and total fat (% BW). Blueberry juice was not as effective as the low dose of anthocyanins in the drinking water in preventing obesity. Additional studies are needed to determine factors responsible for the differing responses of blueberry juice and whole blueberry in preventing the development of obesity.

Urinary excretion of phenolic acids in rats fed cranberry.

Dietary flavonoids can be converted into phenolic acids by colonic microflora. Phenolic acids can then be absorbed into the circulation and may contribute to the health-promoting effects of the parent compounds. Phenolic acids can be further metabolized in other tissues via methylation and conjugation with glucuronide or sulfate. The objectives of this study were to identify and quantify the urinary excretion of 19 phenolic acids and their conjugates in rats fed three levels of a concentrated cranberry powder (3.3, 6.6, and 33 mg/kg of diet). The basic diet used was AIN93G diet containing very low amounts of any polyphenolic compounds. Of the phenolic acids studied, the amounts excreted varied by 4 orders of magnitude, with hippuric acid being excreted in the highest quantities. Amounts of 4-hydroxyphenylacetic acid (4HPAA), 3-hydroxyphenylacetic acid (3HPAA), 3-hydroxyphenylpropionic acid (3HPPA), and 4-hydroxycinnamic acid (4HCA) excreted were in the range of 18-33 microg/mg creatinine in animals fed the highest level of cranberry powder, whereas phenylacetic acid (PAA), gallic acid (GA), 3,4-dihydroxyphenylacetic acid (34HPAA), 3,4-dihydroxybenzoic acid (34HBA), 3,4-dihydroxycinnamic acid (34HCA), and 4-hydroxy-3-methoxycinnamic acid (FA) were excreted in the urine in concentrations of 0.1-2 microg/mg creatinine. As the amount of cranberry in the diet was increased, the amount of 4HPAA excreted decreased but the percentage of conjugated 4HPAA excreted increased (from 57 to 91%). For other phenolic acids analyzed, the percentage excreted in the conjugated form was approximately constant across levels of cranberry in the diet and ranged from 65 to 100% for the individual phenolic acids. Studies of bioactivity and health effects need to consider more than just the compound(s) in the food, because they can be metabolized to other lower molecular weight compounds, which in turn may also be methylated or conjugated in some form that may affect the perceived health effects.

In utero and lactational exposure to blueberry via maternal diet promotes mammary epithelial differentiation in prepubescent female rats.

Early developmental events influence the fine tuning of later susceptibility to adult diseases. Diet is a determinant of breast cancer risk, and our previous studies showed that diet-mediated changes in transcriptional programs promote early mammary gland differentiation. Although consumption of fruits is considered to elicit multiple health benefits, little is known on whether associated bioactive components modify the early differentiation program in developing mammary glands. Here, we evaluated the hypothesis that early exposure (in utero and lactational) to blueberry through maternal diet enhances mammary epithelial differentiation in female offspring. Pregnant Sprague-Dawley rats beginning at gestation day 4 were fed American Institute of Nutrition-based diets containing casein and whole blueberry powders added to casein at 2.5%, 5.0%, and 10% weight/weight. Female pups at weaning were evaluated for growth and mammary tissue parameters. Blueberry at 5% dose increased body and adipose fat weights, relative to the other diets. Mammary branch density and terminal end bud size were highest for the 5% blueberry group, whereas terminal end bud numbers were not affected by all diets. Mammary ductal epithelial cells of the 5% blueberry group had lower nuclear phosphorylated histone 3 and higher nuclear tumor suppressor phosphatase and tensin homolog deleted in chromosome 10 (PTEN) levels than the casein group. Although sera of both diet groups had similar antioxidant capacity, 5% blueberry sera elicited higher nuclear PTEN accumulation in human MCF-10A mammary epithelial cells. Our studies identify developing mammary glands as early targets of blueberry-associated bioactive components, possibly through systemic effects on epithelial PTEN signaling.

Lack of efficacy of blueberry in nutritional prevention of azoxymethane-initiated cancers of rat small intestine and colon.

BACKGROUND: Blueberries may lower relative risk for cancers of the gastrointestinal tract. Previous work indicated an inhibitory effect of consumed blueberry (BB) on formation of aberrant crypt foci (ACF) in colons of male Fisher F344 rats (inbred strain). However, effects of BB on colon tumors and in both genders are unknown. METHODS: We examined efficacy of BB in inhibition of azoxymethane (AOM)-induced colon ACF and intestine tumors in male and female Sprague-Dawley rats (outbred strain). Pregnant rats were fed a diet with or without 10% BB powder; progeny were weaned to the same diet as their dam and received AOM as young adults. RESULTS: Male and female rats on control diet had similar numbers of ACF at 6 weeks after AOM administration. BB increased (P < 0.05) ACF numbers within the distal colon of female but not male rats. There was a significant (P < 0.05) diet by gender interaction with respect to total colon ACF number. Colon and duodenum tumor incidences were less in females than males at 17 weeks after AOM. BB tended (0.1 > P > 0.05) to reduce overall gastrointestinal tract tumor incidence in males, however, tumor incidence in females was unaffected (P > 0.1) by BB. There was a tendency (0.1 > P > 0.05) for fewer adenocarcinomas (relative to total of adenomatous polyps plus adenocarcinomas) in colons of female than male tumor-bearing rats; in small intestine, this gender difference was significant (P < 0.05). BB favored (P < 0.05) fewer adenocarcinomas and more adenomatous polyps (as a proportion of total tumor number) in female rat small intestine. CONCLUSION: Results did not indicate robust cancer-preventive effects of BB. Blueberry influenced ACF occurrence in distal colon and tumor progression in duodenum, in gender-specific fashion. Data indicate the potential for slowing tumor progression (adenomatous polyp to adenocarcinoma) by BB.

Whole berries versus berry anthocyanins: interactions with dietary fat levels in the C57BL/6J mouse model of obesity.

Male C57BL/6J mice received diets with either 10% of calories from fat (LF) or a high-fat diet [45% (HF45) or 60% (HF60) calories from fat] for 92 days (expt 1) or 70 days (expt 2). These were given with or without freeze-dried powders from whole blueberries (BB) or strawberries (SB) (expt 1) or purified anthocyanin extracts from BB or SB (expt 2). Body composition was determined utilizing Echo MRI. Berries added to the LF diet did not alter weight gain, final body weights, body fat, or protein (percent body weight) or diet (grams) or energy (kilocalories) intake. However, in both HF45- and HF60-fed mice, weight gain, final weights, body fat (percent), and epididymal fat weights increased and body protein decreased ( p < 0.01) compared to LF mice. In mice fed HF45 diet plus BB, body weight gains, body fat (percent of BW), and epididymal fat weights were significantly greater than those in the HF45-fed controls, whereas weights of mice fed SB HF were similar to those of HF controls. SB or BB feeding did not alter glucose tolerance, although glucose tolerance decreased with age and in HF45 versus LF mice. Baseline plasma glucose was lower in SB- versus HF45-fed mice. After 8 weeks, mice fed the HF60 diet plus purified anthocyanins from BB in the drinking water had lower body weight gains and body fat than the HF60-fed controls. Anthocyanins fed as the whole blueberry did not prevent and may have actually increased obesity. However, feeding purified anthocyanins from blueberries or strawberries reduced obesity.

Identification and urinary excretion of metabolites of 5-(hydroxymethyl)-2-furfural in human subjects following consumption of dried plums or dried plum juice.

5-(Hydroxymethyl)-2-furfural (I) is a major breakdown product occurring in solutions with high concentrations of fructose and glucose and is present in many fruit juices, in heat-sterilized parenteral solutions, and in baby cereals. The objective of this study was to characterize and identify 5-(hydroxymethyl)-2-furfural metabolites in human subjects following the consumption of dried plum juice and/or dried plums. Subjects were fasted overnight and blood and urine samples were obtained during the day following consumption. Subjects fed the dried plum juice and dried plums consumed 3944 micromol (497 mg) and 531 micromol (67 mg) of I, respectively. Four presumed metabolites of I were detected in the urine of subjects that consumed dried plum juice. They were tentatively identified using HPLC-MS/MS as (1) N-(5-hydroxymethyl-2-furoyl)glycine (III), (2) 5-hydroxymethyl-2-furoic acid (II), (3) (5-carboxylic acid-2-furoyl)glycine (IV), and (4) (5-carboxylic acid-2-furoyl)aminomethane (V). Total urinary excretion during the 6 h following the consumption of dried plum juice was 168, 1465, 137, and 75 micromoles on the basis of II as a standard for II, III, IV, and V, respectively. The estimated total recovery of I metabolites was 46.2% and 14.2% of the I dose during the first 6 h after consumption of dried plum juice and dried plums, respectively. I seems to be metabolized rapidly to glycine conjugates and other metabolites and excreted in the urine.

Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements.

Methods available for the measurement of antioxidant capacity are reviewed, presenting the general chemistry underlying the assays, the types of molecules detected, and the most important advantages and shortcomings of each method. This overview provides a basis and rationale for developing standardized antioxidant capacity methods for the food, nutraceutical, and dietary supplement industries. From evaluation of data presented at the First International Congress on Antioxidant Methods in 2004 and in the literature, as well as consideration of potential end uses of antioxidants, it is proposed that procedures and applications for three assays be considered for standardization: the oxygen radical absorbance capacity (ORAC) assay, the Folin-Ciocalteu method, and possibly the Trolox equivalent antioxidant capacity (TEAC) assay. ORAC represent a hydrogen atom transfer (HAT) reaction mechanism, which is most relevant to human biology. The Folin-Ciocalteu method is an electron transfer (ET) based assay and gives reducing capacity, which has normally been expressed as phenolic contents. The TEAC assay represents a second ET-based method. Other assays may need to be considered in the future as more is learned about some of the other radical sources and their importance to human biology.