Cranberry Fraction Research Articles

Differences in P-Type and Type 1 Uropathogenic Escherichia coli Urinary Anti-Adhesion Activity of Cranberry Fruit Juice Dry Extract Product and D-Mannose Dietary Supplement

Background Urinary tract infection (UTI) prevention benefits of cranberry intake are clinically validated, especially for women and children. To ensure the benefits of cranberry dietary supplement products, the anti-adhesion activity (AAA) against uropathogenic bacteria is routinely used in in vitro bioassays to determine the activity in whole product formulations, isolated compounds, and ex vivo bioassays to assess urinary activity following intake. D-mannose is another dietary supplement taken for UTI prevention, based on the anti-adhesion mechanism. Objective Compare the relative AAA of cranberry and D-mannose dietary supplements against the most important bacterial types contributing to the pathogenesis of UTI, and consider how certain components potentially induce in vivo activity. Methods The current study used a crossover design to determine ex vivo AAA against both P- and Type 1-fimbriated uropathogenic Escherichia coli of either D-mannose or a cranberry fruit juice dry extract product containing 36 mg of soluble proanthocyanidins (PACs), using bioassays that measure urinary activity following consumption. AAA of extracted cranberry compound fractions and D-mannose were compared in vitro and potential induction mechanisms of urinary AAA explored. Results The cranberry dietary supplement exhibited both P-type and Type 1 in vitro and ex vivo AAA, while D-mannose only prevented Type 1 adhesion. Cranberry also demonstrated more robust and consistent ex vivo urinary AAA than D-mannose over each 1-week study period at different urine collection time points. The means by which the compounds with in vitro activity in each supplement product could potentially induce the AAA in urines was discussed relative to the data. Conclusions Results of the current study provide consumers and healthcare professionals with additional details on the compounds and mechanisms involved in the positive, broad-spectrum AAA of cranberry against both E. coli bacterial types most important in UTIs and uncovers limitations on AAA and effectiveness of D-mannose compared to cranberry.

Fractionation of Extracts from Black Chokeberry, Cranberry, and Pomegranate to Identify Compounds That Influence Lipid Metabolism.

Polyphenols show a spectrum of bioactive effects, including an influence on lipid metabolism. In this study, we performed activity-guided fractionations of black chokeberry (aronia), cranberry, and pomegranate extracts to identify the biologically active compounds. The extracts were prepared from fruit juice concentrates with the adsorbent resin Amberlite XAD-7 and were separated into a copigment and an anthocyanin fraction, followed by fractionation into a polymer and monomeric fraction by means of hexane precipitation. For further fractionation of the cranberry and pomegranate copigment fractions, high-performance countercurrent chromatography (HPCCC) was used. The compounds in each fraction were identified by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS), and the quantification was performed by ultra high-performance liquid chromatography-diode array detector (UHPLC-DAD) analyses. Each of the (sub-)fractions was tested in three in vitro assays: phosphodiesterase 3B (PDE) activity, lipid accumulation, and lipolysis in 3T3-L1 cells. The results showed that various fractions and subfractions can inhibit lipid accumulation and PDE activity as well as increase lipolysis, particularly copigments. Overall, our results indicate an influence of polyphenol-rich (sub-)fractions on the lipid metabolism.

An Optimized Fluorometric Method to Test the Capacities of Cranberry Polyphenols and Metabolites to Inhibit the Adhesion of Type-P and Type-1 Fimbriated Uropathogenic E. coli

Abstract Objectives Adhesion of type-P and type-1 fimbriated uropathogenic E. coli to urinary tract epithelial cells initiates urinary tract infections. This research aimed to optimize and apply a fluorometric method to evaluate the capacities of cranberry polyphenols and metabolites to inhibit such adhesion in vitro. Methods BacLight Green labelled E. coli were incubated with cranberry polyphenols or microbial metabolites of cranberry polyphenols for 30 min at 37°C. Mixture was added to a 96-well microplate containing 1 × 105/well of human uroepithelial T24 cells and incubated for 1 h at 37°C. After incubation, E. coli not adhered were removed by phosphate buffer washing. Fluorescent intensity was measured on a microplate reader at 480 nm excitation and 516 nm emission. Results Stable and strong fluorescent readings were obtained with 800 μmol/L BacLight Green for E. coli labeling and an E. coli to T24 cells ratio of 400:1 for co-incubation. A standard curve was established using 0–63 μM myricetin. The half-maximal inhibitory concentrations (IC50) of myricetin were 13.2 μM against type-P E. coli adhesion and 5.5 μM against type-1 E. coli adhesion. A fraction enriched with procyanidin polymers had IC50 of 57.6 μg/mL against type-P E. coli and 19.3 μg/mL against type-1 E. coli, respectively. Its anti-adhesion activities were more potent than those of cranberry fractions enriched with procyanidin oligomers, flavonols, or anthocyanin. Procyanidin A2 had a maximal inhibition about 35% at 17.3 μM against type-P E. coli, but no anti-adhesion activity was observed against type-1 E. coli. Procyanidin B2 showed a plateaued inhibition about 15% at 173–691 μM against type-P E. coli. Its maximal inhibition against type-1 E. coli was around 25% at 346 μM. Hippuric acid, a major metabolite of cranberry polyphenols, had a maximal inhibition about 20% at 558 μM against type-1 E. coli adhesion, whereas its anti-adhesion activity against type-P E. coli was not detected. Conclusions The optimized fluorometric method showed that both structure and composition of cranberry polyphenols and metabolites affected their abilities to inhibit E. coli adhesion in vitro. Anti-adhesion activities of cranberry polyphenols also depend on type of E. coli fimbriae. Funding Sources University of Florida Research Foundation Seed Fund.

Microwave Sensing for Estimating Cranberry Crop Yield: A Pilot Study Using Simulated Canopies and Field Measurement Testbeds

We present the results of an experimental and computational pilot study of cranberry crop yield prediction using low-power microwave sensing and machine learning. We simulated backscattered radiation from cranberry canopies using plane-wave illumination with frequency content from 300 to 2400 MHz. The computational canopy domains allowed for variable soil moisture and cranberry yield in terms of cranberry mass per 1 ft <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sup> of canopy surface area. We collected experimental field data with a prototype open-ended waveguide sensor operating between 600 and 1300 MHz. We measured experimental microwave signals by placing our sensor directly on top of cranberry-crop bed canopies in central Wisconsin and recording reflection coefficients across the operating band. We implemented a machine learning approach to map the microwave reflection coefficients to yield. The mapping procedure involves dimensionality reduction with principal component analysis, supervised learning with linear discriminant analysis, and error-correcting output codes. The idealized computational results demonstrate the feasibility of discriminating three cranberry volume fractions that are representative of central Wisconsin field conditions, at frequencies below 2 GHz. Performance evaluations of the machine learning algorithm applied to the measured field data indicated that, in 81% of test cases, the predicted crop yield had less than 8% error. Most importantly, the average yield prediction error was less than 1.3%. These pilot study results provide strong evidence that machine learning enables accurate cranberry yield estimation when trained with <italic xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">in situ</i> (field) microwave backscattered signals.

Characterization of bioactive cranberry fractions by mass spectrometry

Increasing evidence indicates that fruits contain functional bio-active compounds that have several preventive and therapeutic health benefits. Our group has recently conducted studies to assess the potential effects of cranberry polyphenolic fractions on intestinal Caco-2/15 epithelial cells and a substantial reduction in oxidative stress and inflammation was observed. The aim of the present work was to determine the polyphenolic species most likely responsible for the observed biological activity. Low, medium, and high molecular weight cranberry fractions were generated with a Sephadex LH-20 column by elution with 60% MeOH, 100% MeOH, and 70% acetone, respectively. The total phenolic content in these fractions was determined by the Folin–Ciocalteu method. A combination of LC–MS and MALDI-TOF methods were used to characterize the nature of polyphenolic compounds in the cranberry extracts. High resolution mass spectrometry was used to generate empirical formulae for the detected species. The low molecular weight fraction was essentially constituted of small phenolic acids (hydroxycinnamic and hydroxybenzoic acids). The medium fraction was mostly composed of anthocyanin, flavonols (quercetin, myricetin, isorhamnetin, kaempferol), procyanidins monomers (epicatechin, catechin), dimers, and few small oligomers. The heavy fraction was devoid of phenolic acids and anthocyanins, and it contained for the most part oligomers and polymers of procyanidins. Proanthocyanidins oligomers up to n = 22 were detected, which is, to our knowledge, the largest individual polymers reported to date. With this approach, it was also possible to distinguish between the A-type and B-type linkages.

The synergistic effect of cell wall extracted from probiotic biomass containing Lactobacillus acidophilus CL1285, L. casei LBC80R, and L. rhamnosus CLR2 on the anticancer activity of cranberry juice-HPLC fractions.

Anticancer effects were evaluated on three HPLC fractions obtained from a concentrated cranberry juice and cell wall constituents extracted from a probiotic biomass containing Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, and Lactobacillus rhamnosus CLR2. The samples were tested at increasing concentrations for the antiproliferative assay using HT-29 cells' line and for the quinone reductase (QR) assay using Hepa-1c1c7 murine hepatoma cells. Fraction 1 (F1) which is highly concentrated with phenolic acids inhibited the growth of the HT-29 cells' line with IC50 values of 14.80µg Gallic acid equivalent (GAE)/ml. The fraction 3 (F3) which is highly concentrated in flavonols had potency as QR inducer. Furthermore, the results showed that all cranberry fractions combined with cell wall constituents extracted from the probiotic biomass were more effective in inhibiting the growth of HT-29 as compared to the cranberry fractions tested alone, indicating a possible synergy effect between these bio-functional compounds. PRACTICAL APPLICATIONS: This study strongly evidenced that cranberry juice fractions combined with cell wall constituents extracted from the probiotic biomass can be used as a potent preventive functional compound against colorectal cancer. Therefore, this research proposes a natural dietary compound to prevent mutagenesis and carcinogenesis of colorectal cancer. Furthermore, the industry can formulae products containing probiotic and phenolic compounds as colon cancer cell growth preventive and anticancer products.

Inhibitory Effects of Lingonberry Extract on Oral Streptococcal Biofilm Formation and Bioactivity.

Phenolic compounds in fruits such as cranberries have been shown to promote a number of biological activities. The purpose of this study was to investigate the effects of polyphenolic compound-containing lingonberry extract on oral streptococci and compare them with the known anti-cariogenic activity of cranberries. Water-soluble and polyphenol-rich fractions (Fractions I and II, respectively) were isolated from cranberries and lingonberries. The effects of those fractions on the biofilm formation ability and bioactivity of Streptococcus mutans MT8148R, Streptococcus sobrinus 6715, and Streptococcus sanguinis ATCC 10556 were then evaluated. Cranberry or lingonberry Fraction II (at 0.5-1 mg/ml) significantly reduced biofilm formation by S. mutans, S. sobrinus, and S. sanguinis. In contrast, cranberry or lingonberry Fraction I (at 0.5-2 mg/ml) increased biofilm formation by S. mutans and S. sobrinus, but not by S. sanguinis. Fractions I and II (at 1-2 mg/ml) also reduced the bioactivity of S. mutans, while Fraction II (at 0.5 mg/ml) enhanced the bioactivity of all tested strains. The results revealed that lingonberries contained a larger amount of polyphenol than cranberries and that they showed almost the same level of activity against the biofilm formation ability and bioactivity of oral streptococci. This indicates that polyphenol-rich lingonberry fraction offers a promising natural food derivative for prevention of dental caries.

Effect of cranberry (Vaccinium macrocarpon) oligosaccharides on the formation of advanced glycation end-products.

BACKGROUND: The formation and accumulation of advanced glycation end-products (AGEs) are implicated in several chronic human illnesses including type-2 diabetes, renal failure, and neurodegenerative diseases. The cranberry (Vaccinium macrocarpon) fruit has been previously reported to show anti-AGEs effects, attributed primarily to its phenolic constituents. However, there is lack of similar data on the non-phenolic constituents found in the cranberry fruit, in particular, its carbohydrate constituents. Herein, a chemically characterized oligosaccharide-enriched fraction purified from the cranberry fruit was evaluated for its potential anti-AGEs and free radical scavenging effects.OBJECTIVE: The aim of this study was to evaluate the in vitro anti-AGEs and free radical scavenging effects of a chemically characterized oligosaccharide-enriched fraction purified from the North American cranberry (Vaccinium macrocarpon) fruit.METHOD: The cranberry oligosaccharide-enriched fraction was purified from cranberry hull powder and characterized based on spectroscopic and spectrometric (NMR, MALDI-TOF-MS, and HPAEC-PAD) data. The oligosaccharide-enriched fraction was evaluated for its anti-AGEs and free radical scavenging effects by the bovine serum albumin-fructose, and DPPH assays, respectively.RESULTS: Fractionation of cranberry hull material yielded an oligosaccharide-enriched fraction named Cranf1b-CL. The 1H NMR and MALDI-TOF-MS revealed that Cranf1b-CL consists of oligosaccharides ranging primarily from 6-mers to 9-mers. The monosaccharide composition of Cranf1b-CL was arabinose (25%), galactose (5%), glucose (47%) and xylose (23%). In the bovine serum albumin-fructose assay, Cranf1b-CL inhibited AGEs formation in a concentration-dependent manner with comparable activity to the synthetic antiglycating agent, aminoguanidine, used as the positive control (57 vs. 75%; both at 500μg/mL). In the DPPH free radical scavenging assay, Cranf1b-CL showed superior activity to the synthetic commercial antioxidant, butylated hydroxytoluene, used as the positive control (IC50 = 680 vs. 2200μg/mL, respectively).CONCLUSION: The in vitro anti-AGEs and free radical scavenging effects of cranberry oligosaccharides support previous data suggesting that these constituents may also contribute to biological effects of the whole fruit beyond its phenolic constituents alone. Also, this is the first study to evaluate a chemically characterized oligosaccharide fraction purified from the North American cranberry fruit for anti-AGEs and free radical scavenging properties.

Adherence Reduction of Campylobacter jejuni and Campylobacter coli Strains to HEp-2 Cells by Mannan Oligosaccharides and a High-Molecular-Weight Component of Cranberry Extract

Campylobacter infections are a leading cause of human bacterial gastroenteritis in the United States and are a major cause of diarrheal disease throughout the world. Colonization and subsequent infection and invasion of Campylobacter require that the bacteria adhere to the surface of host cells. Agents that inhibit adherence could be used prophylactically to reduce Campylobacter carriage and infection. Mannan oligosaccharides (MOS) have been used as a feed supplement in livestock animals to improve performance and to replace growth-promoting antibiotics. However, MOS and other nondigestible oligosaccharides may also prevent pathogen colonization by inhibiting adherence in the gastrointestinal tract. In addition, plant extracts, including those derived from cranberries, have been shown to have antiadherence activity against pathogens. The goal of this study was to assess the ability of MOS and cranberry fractions to serve as antiadherence agents against strains of Campylobacter jejuni and Campylobacter coli. Adherence experiments were performed using HEp-2 cells. Significant reductions in adherence of C. jejuni 29438, C. jejuni 700819, C. jejuni 3329, and C. coli 43485 were observed in the presence of MOS (up to 40 mg/ml) and with a high-molecular-weight fraction of cranberry extract (up to 3 mg/ml). However, none of the tested materials reduced adherence of C. coli BAA-1061. No additive effect in adherence inhibition was observed for an MOS-cranberry blend. These results suggest that both components, MOS and cranberry, could be used to reduce Campylobacter colonization and carriage in livestock animals and potentially limit human exposure to this pathogen.

Exploring the role of cranberry polyphenols in periodontits: A brief review.

Cranberry juice polyphenols have gained importance over the past decade due to their promising health benefits. The bioactive component, proanthocyanidins is mainly responsible for its protective effect. A lot has been said about its role in urinary tract infection and other systemic diseases, but little is known about its oral benefits. An extensive search was carried out in the PubMed database using the terms “cranberry polyphenols” and “periodontitis” together. The institute library was also thoroughly scrutinized for all relevant information. Thus, a paper was formulated, the aim of which was to review the role of high molecular weight cranberry fraction on oral tissues and periodontal diseases.

Consumption of cranberry polyphenols enhances human γδ-T cell proliferation and reduces the number of symptoms associated with colds and influenza: a randomized, placebo-controlled intervention study

BackgroundOur main objective was to evaluate the ability of cranberry phytochemicals to modify immunity, specifically γδ-T cell proliferation, after daily consumption of a cranberry beverage, and its effect on health outcomes related to cold and influenza symptoms.MethodsThe study was a randomized, double-blind, placebo-controlled, parallel intervention. Subjects drank a low calorie cranberry beverage (450 ml) made with a juice-derived, powdered cranberry fraction (n = 22) or a placebo beverage (n = 23), daily, for 10 wk. PBMC were cultured for six days with autologous serum and PHA-L stimulation. Cold and influenza symptoms were self-reported.ResultsThe proliferation index of γδ-T cells in culture was almost five times higher after 10 wk of cranberry beverage consumption (p <0.001). In the cranberry beverage group, the incidence of illness was not reduced, however significantly fewer symptoms of illness were reported (p = 0.031).ConclusionsConsumption of the cranberry beverage modified the ex vivo proliferation of γδ-T cells. As these cells are located in the epithelium and serve as a first line of defense, improving their function may be related to reducing the number of symptoms associated with a cold and flu.Trial registrationClinicalTrials.gov Identifier: NCT01398150.

In vitro and in vivo antibacterial activities of cranberry press cake extracts alone or in combination with β-lactams against Staphylococcus aureus

BackgroundCranberry fruits possess many biological activities partly due to their various phenolic compounds; however the underlying modes of action are poorly understood. We studied the effect of cranberry fruit extracts on the gene expression of Staphylococcus aureus to identify specific cellular processes involved in the antibacterial action.MethodsTranscriptional profiles of four S. aureus strains grown in broth supplemented or not with 2 mg/ml of a commercial cranberry preparation (Nutricran®90) were compared using DNA arrays to reveal gene modulations serving as markers for biological activity. Ethanol extracted pressed cakes from fresh fruits also produced various fractions and their effects on marker genes were demonstrated by qPCR. Minimal inhibitory concentrations (MICs) of the most effective cranberry fraction (FC111) were determined against multiple S. aureus strains and drug interactions with β-lactam antibiotics were also evaluated. Incorporation assays with [3H]-radiolabeled precursors were performed to evaluate the effect of FC111 on DNA, RNA, peptidoglycan (PG) and protein biosynthesis.ResultsTreatment of S. aureus with Nutricran®90 or FC111 revealed a transcriptional signature typical of PG-acting antibiotics (up-regulation of genes vraR/S, murZ, lytM, pbp2, sgtB, fmt). The effect of FC111 on PG was confirmed by the marked inhibition of incorporation of D-[3H]alanine. The combination of β-lactams and FC111 in checkerboard assays revealed a synergistic activity against S. aureus including strain MRSA COL, which showed a 512-fold drop of amoxicillin MIC in the presence of FC111 at MIC/8. Finally, a therapeutic proof of concept was established in a mouse mastitis model of infection. S. aureus-infected mammary glands were treated with amoxicillin, FC111 or a combination of both; only the combination significantly reduced bacterial counts from infected glands (P<0.05) compared to the untreated mice.ConclusionsThe cranberry fraction FC111 affects PG synthesis of S. aureus and acts in synergy with β-lactam antibiotics. Such a fraction easily obtained from poorly exploited press-cake residues, may find interesting applications in the agri-food sector and help reduce antibiotic usage in animal food production.

Cranberry bioactives induce TLR signaling and promote a state of immune preparedness in THP‐1 monocytes/macrophages

Cranberries are a rich source of proanthocyanidins (PAC). Because of the unique structure of PAC, we hypothesized that cranberry interacts with pathogen recognition receptors on innate immune cells. Our objective was to evaluate Toll‐like receptor (TLR)‐4 PAC recognition and resulting changes in cytokine mRNA and protein. HEK‐Blue‐hTLR4 cells, containing inducible SEAP reporter genes, were treated with cranberry fractions, LPS, or vehicle. Cranberry fraction containing primarily PAC significantly increased SEAP activity in cell supernatant relative to vehicle. To determine if cranberry primes innate immune cells, THP‐1 cells were differentiated into macrophages and incubated with cranberry or vehicle. Cranberry treatment resulted in significantly higher MIP‐1α mRNA levels without TNFα and IL‐ 1β mRNA increases. An LPS challenge of cranberry‐treated macrophages induced a subtle increase in MIP‐1α protein secretion. In a second experiment, THP‐1 macrophages were treated with cranberry fractions, LPS, or DMSO, then treated with actinomycin D and harvested at times up to 3 hr. Cranberry treatment stabilized MIP‐1α transcripts to a greater extent than did LPS or vehicle. This data suggest that cranberry activates a TLR signaling pathway and primes THP‐1 cells for enhanced response to pathogen challenge via transcript stabilization. Funding provided by Ocean Spray Cranberries, Inc

Inhibition of Streptococcus mutans amyloid fibril formation by cranberry fractions

Biofilm formation is necessary to establish urinary tract infections, dental caries and numerous other infectious conditions. S. mutans is an organism that forms biofilms and was recently shown to be an amyloid‐forming organism. Amyloids are insoluble fibrillar protein aggregates that have been associated with protein malfolding in disease states such as Alzheimer's, but the concept of functional amyloid formation as a directed process involved in microbial pathogenesis is recently emerging. A panel of well‐characterized cranberry fractions (OSC Inc) was tested to determine if they inhibited S. mutans biofilm formation. The fractions did not affect bacterial growth, but inhibited biofilm formation. As a potential mechanism of biofilm inhibition, amyloid fibrillization by S.mutans extracellular proteins was assessed using a Thioflavin‐T fluorescence assay. Amyloid formation was inhibited by cranberry fractions containing proanthocyanidins. Thus, cranberry bioactive compounds may inhibit biofilm formation by preventing amyloid fibril development of the amyloidogenic proteins produced by S. mutans. Support by Univ Schol Prog, UFL, and NIDCR R01DE21789

The effect of American cranberry (Vaccinium macrocarpon) constituents on the growth inhibition, membrane integrity, and injury of Escherichia coli O157:H7 and Listeria monocytogenes in comparison to Lactobacillus rhamnosus

The antimicrobial properties of the American cranberry were studied against Escherichia coli O157:H7, Listeria monocytogenes, and Lactobacillus rhamnosus to determine the effects on growth inhibition, membrane permeability, and injury. Cranberry powder was separated using a C-18 Sep-Pak cartridge into sugars plus organic acids (F1), monomeric phenolics (F2), and anthocyanins plus proanthocyanidins (F3). Fraction 3 was further separated into anthocyanins (F4) and proanthocyanidins (F5) using an LH-20 Sephadex column. Each fraction was diluted in the brain heart infusion (BHI) broth to determine the minimum inhibitory/bactericidal concentrations (MIC/MBC). L. monocytogenes was the most susceptible to cranberry fraction treatment with the lowest MIC/MBC for each treatment, followed by E. coli O157:H7 and L. rhamnosus. Membrane permeability and potential was studied using LIVE/DEAD viability assay and using Bis (1, 3-dibutylbarbituric acid) trimethine oxonol (DiBAC4), respectively. L. rhamnosus demonstrated the highest permeability followed by E. coli O157:H7, and L. monocytogenes. L. rhamnosus demonstrated the highest recovery followed by E. coli O157:H7, and L. monocytogenes. Each cranberry fraction demonstrated membrane hyperpolarization at their native pH, while F2, F3, and F5 demonstrated membrane depolarization at neutral pH. With this knowledge cranberry compounds may be used to prevent maladies and potentially substitute for synthetic preservatives and antibiotics.

Cranberry bioactives augment the activation status of THP‐1 monocytes/macrophages

Cranberries contain proanthocyanidins (PAC), which have been suggested to interact with immune cell pathogen recognition receptors. We hypothesized that recognition of cranberry PAC would alter cellular function toward an activated state. THP‐1 monocytes were incubated with cranberry fractions and analyzed by flow cytometry with FITC‐CD49a. Cranberry fractions activated monocytes in a time and dose dependent manner as shown by increased expression of CD49a on the surface. Cytokines and chemokines secreted in the culture medium were analyzed by multiplex technology. MIP‐1a, MIP‐1β, and IL‐8 were secreted by the cells without marked secretion of pro‐inflammatory cytokines, IL‐1a, IL‐1β, TNF‐a, and IL‐6. Significantly more MIP‐1a mRNA was present. This suggests compounds found in cranberry can activate monocytes to help regulate immune cell trafficking. A second experiment was designed to determine if cranberry primes immune cells. THP‐1cells were differentiated into macrophages and treated with cranberry fractions or DMSO vehicle for 16 hr. Cells were then washed and challenged with LPS for 6 hr. A trend toward increased MIP‐1a protein was found in cranberry‐treated cells compared to vehicle‐treated cells. Taken together, this suggests that cranberry bioactives prime THP‐1 macrophages for a heightened response upon pathogen challenge.Grant Funding Source : Ocean Spray Cranberries, Inc. and AES, University of Florida

Cranberry bioactives promote activation and proliferation of innate immune cells

A previous human intervention study showed that consumption of a cranberry beverage enhanced proliferation of γδ‐T cells and NK cells when they were stimulated ex vivo. To explore potential mechanisms, we asked if we could replicate this observation using in vitro PBMC cultures. Buffy coats were obtained (LifeSouth, Gainesville, Fl) and incubated with several concentrations of cranberry fraction E or F. Cell counts, proliferation and activation of γδ‐T cells, NK cells and NK‐T cells were determined before culture, at 24h and 10d by flow cytometry. In general, all three cell types required more than 24 hours for proliferation to occur. There was a dose response to both cranberry fractions. Cell activation was demonstrated by an increased expression of CD69, an early activation marker on γδ‐T cells and CD314 (NKG2D) on the NK and NK‐T cells. Activation was evident within 24h and was apparent but much lower after 10d. In vivo, only gut immune cells would come into contact with these cranberry polyphenols, however, as one of our first lines of defense; this is an important population to maintain. Supported by Ocean Spray Cranberries, Inc. and AES, UF

Functional Foods: Towards Improving Oral Health

Functional Foods: Towards Improving Oral Health

Cancer chemopreventive effect of fractions from cranberry products

Cancer chemopreventive properties were evaluated in HPLC fractions of different polarity obtained from two cranberry juices and three extracts isolated from frozen cranberries and pomace containing anthocyanins, water-soluble and apolar phenolic compounds, respectively. Compounds with close polarities were collected in order to obtain between three and four fractions from each juice or extract. Cranberry fractions were screened for their ability to induce the phase II xenobiotic detoxification enzyme quinone reductase (QR). The results showed that there was no cytotoxicity against the cells used in the test. All samples stimulated the quinone reductase activity except the highest concentrations of the less polar fraction of anthocyanin-rich extract from pomace, which inhibited the QR activity. The QR induction for all samples varied with the concentration and there was an optimal concentration for which the QR induction was maximal. The technological process to manufacture cranberry juice had little influence on the overall QR inducer potencies of cranberry fractions, whereas the ability of phenols in fractions to stimulate the QR activity has been reduced significantly (P≤0.05) during the technological process. Among all samples, phenolic compounds of eight fractions presented a maximum QR induction greater than 100 II(QR)/mg phenol. The phenolic compounds of the most polar fraction (rich in phenolic acids) and those of the less polar fraction (rich in proanthocyanidins) showed stronger induction than those observed with phenols from intermediate fractions.

Cranberry Phytochemicals Inhibit Glycation of Human Hemoglobin and Serum Albumin by Scavenging Reactive Carbonyls

The objective of this study was to investigate the inhibitory effects of cranberry phytochemicals on protein glycation. Cranberries were purified to yield a sugar‐free powder, which was fractionated into ethyl acetate fraction and water fraction. Water fraction was separated into water fraction‐I, II, and III. Antiglycation activities were tested using glycated hemoglobin (HbA1c), human serum albumin (HSA) ‐glucose, and HSA‐methylglyoxal assays. Cranberry powder and its fractions significantly inhibited the formation of HbA1c by 41.7–81.9 %. The concentrations of cranberry phytochemicals required to inhibit 50% of albumin glycation (EC50) in HSA‐glucose assay was lower than aminoguanidine except for water fraction‐I. The cranberry powder and fractions inhibited glycation of HSA by methylglyoxal. Cranberry phytochemicals scavenged 21.6–89.3% methylglyoxal within 6 hours. Fractions enriched with procyanidins showed higher antiglycation activities. Epicatechin, the constituent unit of procyanidins, were incubated with methylglyoxal and glyoxal at pH 7.4. Five adducts were detected and their structures were tentatively identified using HPLC‐ESI‐MSn. In conclusion, cranberry phytochemicals inhibited glycation of human hemoglobin and serum albumin by scavenging reactive carbonyls. This research is supported in part by Ocean Spray Cranberries, Inc.