Proanthocyanidin-rich Extract Research Articles

Proanthocyanidin-rich extract from Pinus radiata bark: Mild-alkaline extraction and characterization

This study assessed the efficacy of mild-alkaline extraction from P. radiata bark in obtaining proanthocyanidin (PA)-rich extracts. When the bark was treated with three types of bases—Na2CO3, NaHCO3, and NaOH—at varying concentrations, the extract yields increased with higher concentrations. When the pH of the extracts exceeded 7, the PA content and antioxidant activity were remarkably reduced. This result suggests that the pH holds a greater effect in the alkaline extraction of the bark rather than the type of base used. Among the bases used, NaHCO3 was selected and the extraction conditions of pine bark were examined at a concentration where the pH of the extract did not exceed 7. The extraction time during mild-alkaline extraction using 0.2% NaHCO3 was reduced compared to water-only extraction at the same temperature. Moreover, the extract yields were over 10% higher than those of water extraction, and the dried extracts exhibited good solubility in water. The mild-alkaline extracts were characterized using FT-IR and 13C NMR spectroscopic techniques, and acidic alcoholysis. Analyses of the spectra of the mild-alkaline extracts showed similarities to that of pure PA and hot water extract. This result indicated that PA in the bark was not significantly affected during mild-alkaline extraction.

Grape Seed Proanthocyanidins Mitigate the Disturbances Caused by an Abrupt Photoperiod Change in Healthy and Obese Rats.

Variations in the light/dark cycle and obesogenic diets trigger physiological and behavioral disorders. Proanthocyanidins, in addition to their healthy properties, have recently demonstrated a modulating effect on biological rhythms. Therefore, the aim of this study was to evaluate the administration of a grape seed proanthocyanidin-rich extract (GSPE) to mitigate the disruption caused by a sudden photoperiod change in healthy and cafeteria (CAF)-diet obese rats. For this, 48 photoperiod-sensitive Fischer 344 rats were fed standard or CAF diets for 6 weeks under a standard (12 h light/day, L12) conditions. Then, rats were switched to a long (18 h light/day, L18) or short (6 h light/day, L6) photoperiod and administered vehicle or GSPE (25 mg/kg) for 1 week. Body weight (BW) and food intake (FI) were recorded weekly. Animal activity and serum hormone concentrations were studied before and after the photoperiod change. Hormone levels were measured both at 3 h (ZT3) and 15 h (ZT15) after the onset of light. Results showed the impact of the CAF diet and photoperiod on the BW, FI, activity, and hormonal status of the animals. GSPE administration resulted in an attenuation of the changes produced by the photoperiod disruption. Specifically, GSPE in L6 CAF-fed rats reduced serum corticosterone concentration, restoring its circadian rhythm, increased the T3-to-T4 ratio, and increased light phase activity, while under L18, it decreased BW and testosterone concentration and increased the animal activity. These results suggest that GSPE may contribute to the adaptation to the new photoperiods. However, further studies are needed to elucidate the metabolic pathways and processes involved in these events.

Cranberry Proanthocyanidin and Its Microbial Metabolite 3,4-Dihydroxyphenylacetic Acid, but Not 3-(4-Hydroxyphenyl)-Propionic Acid, Partially Reverse Pro-Inflammatory microRNA Responses in Human Intestinal Epithelial Cells.

The molecular basis underlying the anti-inflammatory and anticarcinogenic properties of cranberries is incompletely understood. The effects of a cranberry proanthocyanidin-rich extract (PAC) and two of its gut microbial metabolites, 3,4-dihydroxyphenylacetic acid (DHPAA) and 3-(4-hydroxyphenyl)-propionic acid (HPPA), on intestinal epithelial cells microRNA (miRNA) expression and their downstream pathways at homeostasis and in inflammatory conditions, are investigated. The expression of 799 miRNAs is quantitatively assessed in differentiated Caco-2BBe1 cells pre-treated with PAC, DHPAA, or HPPA and stimulated with interleukin (IL)-1β or not. PAC, DHPAA, and HPPA generate subsets of shared and distinct miRNA responses. At homeostasis, miRNAs affected by the metabolites, but not PAC, targeted genes enriched in kinase, Wnt, and growth factor signaling, cell growth and proliferation, apoptosis, and specific cancer pathways. In an inflammatory environment, PAC and DHPAA, but not HPPA, reverses the expression of 16 and two IL-1β-induced miRNAs, respectively, regulating inflammatory and cancer pathways. miRNA modulation is a novel mechanism for PAC bioactivity in the gut. The gut microbiota may be necessary to unlock these effects at homeostasis and partially in inflammation.

Polyphenol-induced improvements in glucose metabolism are associated with bile acid signaling to intestinal farnesoid X receptor.

IntroductionBile acid (BA) biotransformation by gut bacteria impacts BA profile and signaling to nuclear receptors, such as the farnesoid X receptor (FXR) regulating glucose metabolism. Altered BA-FXR signaling was therefore investigated as a potential mechanism linking polyphenol-induced gut bacterial changes and improved glucose metabolism.Research design and methodsDiabetic db/db were fed low-fat diet (LFD) or LFD supplemented with a proanthocyanidin-rich extract of grape polyphenols (LFD-GP) for 4 weeks. Metabolic phenotypes, serum BAs, gut microbiota composition, and gene expression markers relevant to gut barrier and glucose metabolism were assessed. Gut organoids were used to investigate effects of individual BAs on ileal FXR activity.ResultsCompared with LFD-fed controls, GP supplemented db/db mice showed improved glucose metabolism, decreased relative abundance of gut bacteria associated with production of secondary BAs (SBAs), and depleted serum levels of SBAs taurohyodeoxycholic acid (THDCA), ω-muricholic acid (ωMCA), and tauro-ω-muricholic acid (TωMCA). Serum levels of primary BAs (PBAs) increased, consistent with higher gene expression of PBA synthesis enzyme Cyp7a1. GP-induced BA changes associated with FXR inhibition as evidenced by reduced expression of FXR-responsive genes Shp, Fgf15, and Fabp6 in ileum tissue as well as hepatic Shp, which negatively regulates PBA synthesis. GP treatment did not affect expression of hepatic Fxr or expression of Abcb11, Slc51b, and Obp2a genes controlling BA transport. Ceramide biosynthesis genes Smpd3, Sptlc2, and Cers4 were decreased in liver and intestine suggesting lower tissue ceramides levels may contribute to improved glucose metabolism. THDCA, ωMCA, and TωMCA behaved as FXR agonists in ileal organoid experiments; therefore, their depletion in serum of GP-supplemented db/db and wild type (WT) mice was consistent with FXR inhibition.ConclusionThese data suggest that by altering the gut microbiota, GPs modify BA-FXR signaling pathways to promote glucoregulation.

The effects of Vitis vinifera L. phenolic compounds on a blood-brain barrier culture model: Expression of leptin receptors and protection against cytokine-induced damage.

The medicinal properties of grapes (Vitis vinifera L.) are well known since ancient times. Ethnobotanical grape preparations, like the Ayurvedic Darakchasava are used as cardiotonic and for the treatment of cardiovascular diseases. Dried grape products are also applied in Iranian traditional medicine for memory problems, which are linked to the pathology of brain microvessels, a special part of the cardiovascular system. The anti-inflammatory and protective effects of these traditional preparations on the cardiovascular system are related to their bioactive phenolic compounds. The blood-brain barrier (BBB), formed by brain capillaries, is not only involved in inflammatory and other diseases of the central nervous system, but also in many systemic diseases with an inflammatory component. Dietary obesity is a systemic chronic inflammatory condition in which the peripheral and central vascular system is affected. Among the cerebrovascular changes in obesity defective leptin transport across the BBB related to central leptin resistance is observed. Our aim was to study the protective effects of grape phenolic compounds epicatechin (EC), gallic acid (GA) and resveratrol (RSV) and grape-seed proanthocyanidin-rich extract (GSPE) on a cytokine-induced vascular endothelial inflammation model. Using a culture model of the BBB we investigated cytokine-induced endothelial damage and changes in the expression of leptin receptors and leptin transfer. For the BBB model, primary cultures of rat brain endothelial cells, glial cells and pericytes were used in co-culture. Cells were treated by tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β) (10 ng/ml each) to induce damage. Cell toxicity was evaluated by the measurement of impedance. The expression of leptin receptors was assessed by RT-qPCR and western blot. The production of reactive oxygen species (ROS) and nitric oxide (NO) were detected by fluorescent probes. GSPE (10 μg/ml), EC (10 μM), GA (1 μM) or RSV (10 μM) did not change the viability of brain endothelial cells. The gene expression of the short leptin receptor isoform, Ob-Ra, was up-regulated by GSPE, EC and RSV, while the mRNA levels of Lrp2 and clusterin, clu/ApoJ were not affected. The tested compounds did not change the expression of the long leptin receptor isoform, Ob-Rb. RSV protected against the cytokine-induced increase in albumin permeability of the BBB model. GSPE and EC exerted an antioxidant effect and GSPE increased NO both alone and in the presence of cytokines. The cytokine-induced nuclear translocation of transcription factor NF-κB was blocked by GSPE, GA and RSV. Cytokines increased the mRNA expression of Lrp2 which was inhibited by EC. RSV increased Ob-Ra and Clu in the presence of cytokines. Cytokines elevated leptin transfer across the BBB model, which was not modified by GSPE or RSV. Our results obtained on cell culture models confirm that natural grape compounds protect vascular endothelial cells against inflammatory damage in accordance with the ethnopharmacological use of grape preparations in cardiovascular diseases. Furthermore, grape compounds and GSPE, by exerting a beneficial effect on the BBB, may also be considered in the treatment of obesity after validation in clinical trials.

Rapid and selective mobilization of specific stem cell types after consumption of a polyphenol-rich extract from sea buckthorn berries (Hippophae) in healthy human subjects.

PurposeThe aim of this study was to evaluate the effects of a proanthocyanidin-rich extract of sea buckthorn berry (SBB-PE) on the numbers of various types of adult stem cells in the blood circulation of healthy human subjects.Study design and methodsA randomized, double-blind, placebo-controlled, cross-over trial was conducted in 12 healthy subjects. Blood samples were taken immediately before and at 1 and 2 hours after consuming either placebo or 500 mg SBB-PE. Whole blood was used for immunophenotyping and flow cytometry to quantify the numbers of CD45dim CD34+ CD309+ and CD45dim CD34+ CD309− stem cells, CD45− CD31+ CD309+ endothelial stem cells, and CD45− CD90+ mesenchymal stem cells.ResultsConsumption of SBB-PE was associated with a rapid and highly selective mobilization of CD45dim CD34+ CD309− progenitor stem cells, CD45− CD31+ CD309+ endothelial stem cells, and CD45− CD90+ lymphocytoid mesenchymal stem cells. In contrast, only minor effects were seen for CD45dim CD34+ CD309+ pluripotential stem cells.ConclusionConsumption of SBB-PE resulted in selective mobilization of stem cell types involved in regenerative and reparative functions. These data may contribute to the understanding of the traditional uses of SBB for preventive health, regenerative health, and postponing the aging process.

Effect of natural collagen cross-linker concentration and application time on collagen biomodification and bond strengths of fiber posts to root dentin

Proanthocyanidin-rich plant-derived agents have been shown to increase dentin biomechanical properties and resistance of dentinal collagen to degradation. This study investigated the effects of proanthocyanidin-rich extract (grape seed extract, GSE; Vitis vinifera) on the mechanical properties of demineralized dentin matrix, and the bond strength and stability of the adhesion of fiber posts to the root dentin. Demineralized dentin beams (n = 15) were incubated with 6.5% or 10% GSE for 30, 60, or 120 s. The modulus of elasticity (E) and the fold increase in E were determined by comparing specimens at baseline and after treatment. The data were analyzed by two-way ANOVA and Tukey post hoc tests (α = 0.05). For the bond strength test, single-rooted human teeth were randomly divided into seven groups: control group, no treatment; and experimental groups as described for the modulus of elasticity. Fiber posts were cemented with RelyX U200. Each group was randomly divided into two subgroups: 24 hours of water storage and 12 months of water storage. All roots were sectioned transversely in the coronal, middle, and apical regions, producing 1-mm thick slices, and the push-out test was performed. The failure modes were observed, and the bond strength means were analyzed by ANOVA (α = 0.05). For E test, no significant difference was observed among the incubation times (P > 0.05); however, high concentration of GSE (10%) resulted in a statistically higher fold increase in E compared to 6.5% (P < 0.05). For the push out test, the immediate and stored for 12 months groups showed similar bond strength values with or without pretreatment with GSE (P > 0.05), irrespective of the GSE concentration, time of treatment, and root regions (P > 0.05); however, the use of GSE contributed to a significant reduction in adhesive failure mode. The results indicate that 10% GSE exhibits a high cross-linking potential for dentin extracellular matrix. Moreover, the bond strength of fiber posts luted to root dentin with RelyX U200 remained stable after 12 months of water storage, with or without pre-treatment with GSE.

Skin Anti-aging Assays of Proanthocyanidin Rich Red Rice Extract, Oryzanol and Other Phenolic Compounds

Red rice is a variety of rice that has more nutritious than white or brown rice. It is also a good source of many potent anti-aging phytochemicals. However, the compounds in red rice extract that exhibit skin anti-aging properties have not been investigated. In this study, the main bioactive compounds in red rice extract (RRE) including proanthocyanidin, catechin, hydroxybenzoic acid, vanillic acid and oryzanol were studied in order to determine their anti-skin aging properties. The effects on skin degradation were assessed by inhibitory enzymatic activity against collagenase and matrixmetalloproteinase-2 (MMP-2). The production levels of collagen and hyaluronic acid obtained from human skin fibroblasts were determined by ELISA. Anti-melanogenesis activity of the bioactive compounds were investigated in B16-F10 mouse melanoma cells. The activity of collagenase and MMP-2 was strongly inhibited by proanthocyanidin and catechin, while hydroxybenzoic acid, vanillic acid and oryzanol had no effect. Moreover, proanthocyanindin and catechin significantly induced collagen and hyaluronic acid synthesis in human fibroblast cells. Proanthocyanidin and oryzanol reduced the melanin content in B16-F10 mouse melanoma cells. Proanthocyanidin, but not oryzanol, significantly decreased cellular tyrosinase activity. However, the bioactive compounds obtained from red rice extract had no effect on mushroom tyrosinase activity. In addition, proanthocynidin and catechin, exhibited strong DPPH radical scavenging activity, whereas oryzanol slightly inhibited this action. Taken together, these results suggest that proanthocyanidin, catechin, and oryzanol are the bioactive compounds in red rice that exhibit the greatest levels of anti-skin aging properties.

Development and validation of an UHPLC-HRMS protocol for the analysis of flavan-3-ol metabolites and catabolites in urine, plasma and feces of rats fed a red wine proanthocyanidin extract

This study developed, optimized and validated an ultra-high-performance liquid chromatography–high-resolution mass spectrometry (UHPLC-HRMS) method to identify and quantify metabolites and microbial-derived catabolites in urine, plasma and feces of rats following ingestion of 50 mg of a red wine proanthocyanidin-rich extract. The method was validated for specificity, linearity, limit of detection (LD) and quantification (LQ), intra-day and inter-day precision, recovery and matrix effects, which were determined for 34 compounds in the three biological matrices. After method validation, three parent flavan-3-ols, four 5-carbon side chain ring fission metabolites, and 27 phenolic acid and aromatic catabolites were quantified in plasma, urine and feces after red wine proanthocyanidin intake. These results establish the value of the UHPLC-HRMS protocol in obtaining a detailed picture of proanthocyanidin metabolites and their microbial-derived catabolites, along with their phase II metabolites, in biological fluids of rat, and potentially in human clinical studies designed to evaluate the bioavailability of dietary flavan-3-ols.

A proanthocyanidin-rich extract from Cassia abbreviata exhibits antioxidant and hepatoprotective activities in vivo

Ethnopharmacological relevanceCassia abbreviata is a small to medium sized branched umbrella-shaped deciduous tree. It is widely spread in the tropics, especially in Africa, having a long history in traditional medicine for the treatment of numerous conditions such as headaches, diarrhea, constipation, some skin diseases, malaria, syphilis, pneumonia, stomach troubles, uterine pains, and against gonorrhea. Aim of the studyWe investigated the phytochemical constituents of a root extract from Cassia abbreviata using HPLC-PDA-ESI-MS/MS. We also determined the antioxidant activities in vitro and in vivo using the nematode Caenorhabditis elegans as a model organism. The hepatoprotective activities in case of D-galactosamine (D-GaIN)-induced hepatotoxicity were studied in a rat model. Materials and methodsHPLC-PDA-ESI-MS/MS analysis allowed the identification of the secondary metabolites of the methanol extract. DPPH and FRAP assays were used to determine the antioxidant activities in vitro. Using the C. elegans model, survival rates under juglone induced oxidative stress, intracellular ROS content, quantification of Phsp-16.2: GFP expression and subcellular DAF-16: GFP localization were investigated to determine the antioxidant activities in vivo. The in vivo hepatoprotective potential of the root extract was evaluated for D-galactosamine (D-GaIN)-induced hepatotoxicity in rats. The activity of the liver enzymes alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyltransferase (GGT), in addition to liver peroxidation product malondialdehyde (MDA) and glutathione content (GSH), as well as albumin and total bilirubin concentration, were determined. A histopathological study was also performed. Results and conclusionsC. abbreviata root extract is rich in polyphenolics, particularly proanthocyanidins. HPLC-PDA-ESI-MS/MS analysis resulted in the identification of 57 compounds on the bases of their mass spectra. (epi)-Catechin, (epi)-afzelechin, (epi)-guibourtinidol, and (ent)-cassiaflavan monomers as well as their dimers, trimers, and their diastereomers are the main components of the extract.The total phenolic content amounted for 474mg/g root extract expressed as gallic acid equivalent using the Folin-Ciocalteu method. The extract exhibited powerful antioxidant activity with EC50 of 6.3μg/mL in DPPH and 19.15mM FeSO4 equivalent/mg sample in FRAP assay. In C. elegans model, the extract (200μg/mL) was able to increase the survival rate by 44.56% and reduced the ROS level to 61.73%, compared to control group.Pretreatment of rats with 100mg extract/kg (b. wt.) reduced MDA by 47.36% and elevated GSH by 59.1%. The extract caused a significant reduction of ALT, AST and GGT activities by 11%, 35.7% and 65%, respectively.The findings of this study suggest that the proanthocyanidin-rich extract from C. abbreviata may be an interesting candidate for hepatoprotective activity in case of hepatocellular injury.

Great diversity among commercial inactive dry-yeast based products

Commercial inactive dry-yeast based (IDYB) products have been shown to impact positively in different ways on the winemaking process, including sensory enhancement. Despite their relevance little information about physicochemical characteristics of individual IDYB products is available. This study aimed to physicochemically characterize a group of ten commercial IDYB products. Organic, protein and carbohydrate contents by spectrophotometric methods, protein diffusion on cellulose membranes and electrophoretic protein profiles were assessed. Interaction of a IDYB product (CP10) with either salivary protein or a proanthocyanidin-rich extract (binary mixtures) or with both of them (ternary mixtures) was also assessed. Marked physicochemical differences were observed among all ten products. CP10 was found to interact with seed extract and salivary protein. Also, as part of CP10-SE complexes, CP10 interacted with the salivary protein to form ternary complexes. Due to their huge diversity, physicochemical characterization of IDYB products before use in winemaking is recommended.

Cranberry proanthocyanidins modulate reactive oxygen species in Barrett's and esophageal adenocarcinoma cell lines.

BACKGROUND: We recently reported that a cranberry proanthocyanidin rich extract (C-PAC) induces autophagic cell death in apoptotic resistant esophageal adenocarcinoma (EAC) cells and necrosis in autophagy resistant cells. EAC is characterized by high morbidity and mortality rates supporting development of improved preventive interventions.OBJECTIVE: The current investigation sought to investigate the role of reactive oxygen species (ROS) in the context of C-PAC induced cell death.METHODS: A panel of human esophageal cell lines of EAC or BE (Barrett’s esophagus) origin were treated with C-PAC and assessed for ROS modulation using CellROX® Green reagent and the Amplex Red assay to specifically measure hydrogen peroxide levels.RESULTS: C-PAC significantly increased ROS levels in EAC cells, but significantly reduced ROS levels in CP-C BE cells. Increased hydrogen peroxide levels were also detected in C-PAC treated EAC cells and supernatant; however, hydrogen peroxide levels were significantly increased in medium alone, without cells, suggesting that C-PAC interferes or directly acts on the substrate. Hydrogen peroxide levels did not change in C-PAC treated CP-C BE cells.CONCLUSION: These experiments provide additional mechanistic insight regarding C-PAC induced cancer cell death through modulation of ROS. Additional research is warranted to identify specific ROS species associated with C-PAC exposure.

Bond Strength between Fiber Posts and Root Dentin Treated with Natural Cross-linkers

IntroductionThe aim of this study was to investigate the effects of a proanthocyanidin-rich extract (grape seed extract [GSE]) on the bond strength and stability of the adhesion of fiber posts to the root dentine using 2 adhesive systems: a total-etch and a self-etch adhesive system. MethodsSingle-rooted human teeth were randomly divided into 6 groups: G1 (control), untreated + total-etch adhesive Adper Scotchbond Multi-Purpose (3M ESPE, St Paul, MN); G2, 6.5% GSE for 5 minutes + SB; G3, 10% GSE for 5 minutes + SB; and G4 to G6 groups were similar to previous ones; however, the self-etch adhesive system Clearfil SE Bond (Kuraray, Kurashiki, Japan) was used. Fiber posts were cemented with RelyX ARC (3M ESPE), and the specimens were immediately tested for push out or stored for 12 months. The bond strength means were analyzed by analysis of variance and Games-Howell post hoc tests (α = 0.05). Additionally, matrix metalloproteinase inhibition by 0.65%, 0.065%, and 0.0065% GSE was examined with gelatin zymography. ResultsThe use of GSE did not affect immediate bond strength to dentin and contributed to preserve the bond strength after 12 months (P < .05) for both adhesives. The bond strength of SB was significantly higher than Clearfil SE Bond (P < .05). Gelatin zymography showed reduced matrix metalloproteinase activity when recombinant enzymes were incubated with both 0.65% and 0.065% GSE, with complete inhibition at the highest concentration. ConclusionsThe pretreatment with GSE can be used as a natural alternative to improve bond strength stability of dentin–adhesive interfaces in root canals.

Serum metabolites of proanthocyanidin-administered rats decrease lipid synthesis in HepG2 cells

The regular consumption of flavonoids has been associated with reduced mortality and a decreased risk of cardiovascular diseases. The proanthocyanidins found in plasma are very different from the original flavonoids in food sources. The use of physiologically appropriate conjugates of proanthocyanidins is essential for the in vitro analysis of flavonoid bioactivity.In this study, the effect of different proanthocyanidin-rich extracts, which were obtained from cocoa (CCX), French maritime pine bark (Pycnogenol extract, PYC) and grape seed (GSPE), on lipid homeostasis was evaluated. Hepatic human cells (HepG2 cells) were treated with 25 mg/L of CCX, PYC or GSPE. We also performed in vitro experiments to assess the effect on lipid synthesis that is induced by the bioactive GSPE proanthocyanidins using the physiological metabolites that are present in the serum of GSPE-administered rats. For this, Wistar rats were administered 1 g/kg of GSPE, and serum was collected after 2 h. The semipurified serum of GSPE-administered rats was fully characterized by liquid chromatography tandem triple quadrupole mass spectrometry (LC–QqQ/MS2). The lipids studied in the analyses were free cholesterol (FC), cholesterol ester (CE) and triglycerides (TG).All three proanthocyanidin-rich extracts induced a remarkable decrease in the de novo lipid synthesis in HepG2 cells. Moreover, GSPE rat serum metabolites reduced the total percentage of CE, FC and particularly TG; this reduction was significantly higher than that observed in the cells directly treated with GSPE. In conclusion, the bioactivity of the physiological metabolites that are present in the serum of rats after their ingestion of a proanthocyanidin-rich extract was demonstrated in Hep G2 cells.

Immunomodulatory effect of a proanthocyanidin-rich extract from Pinus radiata bark by dosing period in chickens

We evaluated the immunomodulatory effects of a proanthocyanidin-rich extract (PAE) from Pinus radiata bark in Korean native chickens. In experiment 1, proliferation of peripheral blood mononuclear cells was significantly enhanced in chickens administered 1.25, 2.5, 5, and 10 mg/kg of PAE for 2 wk once daily by oral gavage compared with no administration of PAE in vitro. Proliferation of splenocyte and thymocyte cells was also significantly enhanced in chickens administered 1.25, 2.5, 5, and 10 mg/kg of PAE for 5 wk. In experiment 2, proliferation of peripheral blood mononuclear cells, splenocytes, and thymocytes in chickens administered PAE for 6 wk was continuously higher than that in other groups. Additionally, increasing cell populations for which CD4+ CD8−(Th cells) and Bu-1+ (B cells) protein was expressed on the cell surface were significantly different after 4 wk of PAE administration in chickens. Thus, the immunomodulatory effects of a PAE from P. radiata bark differed by dosing period in chickens.

Effect of proanthocyanidin-rich extract from Pinus radiata bark on immune response of specific-pathogen-free White Leghorn chickens

Proanthocyanidins are naturally occurring compounds that are widely found in fruits, vegetables, nuts, seeds, flowers, and bark. We evaluated the immunomodulatory effects of proanthocyanidin-rich extract (PAE) from Pinus radiata bark in specific-pathogen-free White Leghorn chickens. Proliferation of peripheral blood mononuclear cells was significantly enhanced in chickens treated for 2 wk with 20 mg/kg of PAE. Proliferation of splenocytes and bursal cells was significantly enhanced in chickens treated for 5 wk with 5, 10, and 20 mg/kg of PAE. Thymocyte proliferation was significantly enhanced in chickens treated for 5 wk with 5 and 10 mg/kg of PAE. These effects were markedly enhanced by the presence of lipopolysaccharide, which acted on B cells responsible for humoral immunity, and concanavalin A, which acted directly on T cells involved in cell-mediated immunity. The PAE significantly promoted the expression of T helper 1 cytokine (interferon-γ) and decreased the expression of T helper 2 cytokine (IL-6). Thus, P. radiata PAE has immunomodulatory effects in specific-pathogen-free White Leghorn chickens.

MicroRNA alterations in Barrett′s esophagus, esophageal adenocarcinoma, and esophageal adenocarcinoma cell lines following cranberry extract treatment: Insights for chemoprevention

Background:Aberrant expression of small noncoding endogenous RNA molecules known as microRNAs (miRNAs) is documented to occur in multiple cancer types including esophageal adencarcinoma (EAC) and its only known precursor, Barrett's esophagus (BE). Recent studies have linked dysregulation of specific miRNAs to histological grade, neoplastic progression and metastatic potential.Materials and Methods:Herein, we present a summary of previously reported dysregulated miRNAs in BE and EAC tissues as well as EAC cell lines and evaluate a cranberry proanthocyanidin rich extract's (C-PAC) ability to modulate miRNA expression patterns of three human EAC cell lines (JHEso-Ad-1, OE33 and OE19).Results:A review of 13 published studies revealed dysregulation of 87 miRNAs in BE and EAC tissues, whereas 52 miRNAs have been reported to be altered in BE or EAC cell lines, with 48% overlap with miRNA changes reported in tissues. We report for the first time C-PAC–induced modulation of five miRNAs in three EAC cell lines resulting in 26 validated gene targets and identification of key signaling pathways including p53, angiogenesis, T-cell activation and apoptosis. Additionally, mutiple cancer related networks were ideintified as modulated by C-PAC utilizing Kyoto Encyclopedia of Genes and Genomes (KEGG), Protein Analysis Through Evolutionary Relationships (PANTHER), and MetaCore analysis tools.Conclusions:Study results support the cancer inhibitory potential of C-PAC is in part attributable to C-PAC's ability to modify miRNA profiles within EAC cells. A number of C-PAC–modulated miRNAs have been been identified as dysregulated in BE and EAC. Further insights into miRNA dysregulation and modulation by select cancer preventive agents will support improved targeted interventions in high-risk cohorts.

Effects of Proanthocyanidin-rich Extract from Pinus radiata Bark on Immune Responses of Broiler Chickens

Proantocyanidins are naturally occurring compounds that are widely found in fruits, vegetables, nuts, seeds, flowers and bark. In part I, we evaluated the immunomodulatory effects of proanthocyanidin-rich extract (PAE) from Pinus radiata bark in specific pathogen free (SPF) White Leghorn chickens. Proliferation of peripheral blood mononuclear cells was significantly enhanced in chickens treated for 2 weeks with 20 ㎎/㎏ PAE. Proliferation of splenocytes and bursal cells was significantly enhanced in chickens treated for 5 weeks with 5, 10, and 20 ㎎/㎏ PAE. Thymocyte proliferation was significantly enhanced in chickens treated for 5 weeks with 5 and 10 ㎎/㎏ PAE. These effects were markedly enhanced by the presence of lipopolysaccharide, which actedon B cells responsible for humoral immunity, and concanvalin A, which acteddirectly on T cells involved in cell mediated immunity. PAE significantly promoted the expression of Th1 cytokine (interferon-gamma) and decreased the expression of Th2 cytokine (interleukin-6). In part Ⅱ, we evaluated the immunomodulatory effects of proanthocyanidin-rich extract (PAE) from Pinus radiata bark in broiler. Proliferation of peripheral blood mononuclear cells and thymocytes was significantly enhanced in 2.5, 5, 10 ㎎/㎏ PAE-treated broiler chickens. Proliferation of splenocytes was significantly enhanced in 1.25, 2.5, 5, 10 ㎎/㎏ PAE-treated broiler chickens. These effects were markedly enhanced by the presence of LPS, which acts on B cells responsible for humoral immunity, and Con A, which acts directly on T cells involved in cell mediated immunity. PAE significantly promoted the expression of interleukin-18 and interleukin-1 β. In part Ⅲ, We evaluated the immunomodulatory effects of proanthocyanidin-rich extract (PAE) from Pinus radiata bark in Korean native chickens. In experiment 1, proliferation of peripheral blood mononuclear cells (PBMCs) was significantly enhanced in chickens administered for 2 weeks with 1.25, 2.5, 5 and 10 ㎎/㎏ PAE. Proliferation of splenocytes and thymocytes cells was significantly enhanced in chickens administered for 5 weeks with 1.25, 2.5, 5 and 10 ㎎/㎏ PAE. In experiment 2, proliferation of PBMCs, splenocytes and thymocytes in the PAE-administered chickens for 6 weeks were continuously higher than other groups and significantly promoted the expression of Th1 cytokines (IL-2, IL-12, and IFN- γ). Also, the CD4+CD8? (Th1 cell) and Bu-1+ (B cell) cell population significantly upregulated after 4 weeks in PAE-administered chickens.

Abstract B67: In vitro and in vivo inhibitory effects of cranberry proanthocyanidins against esophageal adenocarcinoma

Abstract This study investigated the cancer inhibitory potential of a proanthocyanidin rich cranberry extract utilizing a panel of authenticated human esophageal adenocarcinoma cell lines (EAC), a nude mouse xenograft model, and the rat esophagogastrodoudenal anastomosis (EGDA) model of EAC. PACs effects on cell morphology, global gene expression, phase of cell cycle, and cell death induction via apoptosis, autophagy, and necrosis were evaluated. Methods included RapidDiff and MDC staining, Annexin/PI and BrdU staining with flow cytometric evaluation, transmission electron microscopy (TEM), protein evaluation utilizing standard Western blot techniques, and validation of global gene expression changes by real-time PCR. PAC treatment [50-100µg/ml] of EAC cancer cell lines resulted in significant increases in apoptosis, autophagy and necrosis. Differential cell death induction was noted based upon the cell lines resistance to a bile/acid cocktail mimicking acid exposure in humans as well as the status of the apoptotic and autophagic machinery of the select cell line. PACs cell death inducing capacity correlated with altered MAPKinase signaling as evidenced by activation of P-p38 and P-JNK and inhibition of P-ERK1/2. PAC treatment induced other proapoptotic markers including cleaved PARP, BAK1, BAX, and cytochrome C, but in a cell line dependent manner. Mild increases in Beclin-1 were noted following PAC treatment, as were significant increases in LC3II, a marker of autophagasome formation which occurs during autophagy induction. Results were confirmed by TEM and MDC staining. PAC induced cell cycle arrest at the G2/M transition, increased the percent of cells in G1, caused an S-phase delay and increased P21 levels in all three cell lines. PAC administration significantly inhibited the growth of OE19 xenografts via modulation of cell cycle and MAPK signaling pathways. Oral gavage of PAC [250µg/mouse] six times a week to nude mice resulted in a 67% decrease in OE19 tumor volume, increased cytochrome C protein levels and decreased levels of P-ERK1/2, cyclin A, PCNA, and CD31 in PAC treated tumors compared to vehicle treated tumors. Lastly, a four week pilot study was performed to evaluate the safety of daily administration of PAC in the rodent EGDA model. PAC appeared well tolerated given the average daily dose of 450µg/rat/day as evidenced by normal body weight and food consumption, serology profiles and histopathology of the major organs. Further long-term studies are planned to more fully access the chemopreventive potential of PAC against esophageal adenocarcinoma and premalignancy utilizing the clinically relevant rat EGDA surgical model. Citation Information: Cancer Prev Res 2010;3(12 Suppl):B67.

Apple Proanthocyanidins Do Not Reduce the Induction of Preneoplastic Lesions in the Colon of Rats Associated with Human Microbiota

Since the gut microbiota metabolizes various dietary constituents unabsorbed by the small intestine and modulates colon function, it plays an essential role in colon carcinogenesis. First, we have developed a model of human microbiota-associated rats (HMA), fed a human-type diet and injected with 1-2,dimethylhydrazine (DMH). We observed that the number and size of DMH-induced aberrant crypt foci (ACF) were significantly higher in HMA rats than in germ-free or conventional rats. Second, we used this model to assess the protective effect of an apple proanthocyanidin-rich extract (APE) on colon carcinogenesis. In this model, ACF number and multiplicity were not reduced by APE at 0.001% and 0.01% in drinking water. They were higher with APE 0.1% than with APE 0.01%. Therefore, the cross-talk between human microbiota and the colon epithelium should be taken into account in carcinogenesis models. Moreover, attention should be paid prior to using proanthocyanidin extracts as dietary supplements for humans.